Apelin在多囊卵巢综合征中表达及对颗粒细胞作用机制的初步探讨
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摘要
多囊卵巢综合征(polycystic ovary syndrome，PCOS)是生育期女性常见的一种内分泌紊乱伴代谢异常疾病，发病率4-10%，以稀发排卵或持续无排卵、高雄激素血症、胰岛素抵抗（IR）和卵巢多囊样改变为特征。PCOS患者近期表现为月经失调、不孕、多毛、痤疮、肥胖，远期心血管疾病、糖尿病、肿瘤发生风险明显增高，PCOS严重威胁患者的生理及心理健康。生育期PCOS女性排卵异常是月经稀发或闭经、不孕的最主要原因。研究PCOS患者卵泡发育，有利于了解其生理病理改变及其可能的发病机制。PCOS患者卵巢内的始基卵泡数量与正常健康妇女比较无明显差异，而窦状卵泡、闭锁卵泡数量明显增多，无优势卵泡发育，导致排卵障碍，影响月经、妊娠及妊娠结局等。有学者提出PCOS是原发性卵泡病，而颗粒细胞的增殖、凋亡是卵泡发育及闭锁的根本原因，研究发现卵巢局部众多细胞因子通过旁分泌或者自分泌等方式参与颗粒细胞增殖或凋亡。
无排卵PCOS患者改变生活方式后，排卵恢复者的体重、腹部脂肪减少比未恢复者更明显；此外PCOS患者内脏脂肪持续减少与排卵恢复紧密相关。可见肥胖及IR相关脂肪因子在PCOS患者卵泡发育中有重要影响，近年大量国内外研究侧重于探讨脂肪因子如脂联素、瘦素等在PCOS患者基因表达和（或）蛋白功能缺陷对脂、糖代谢影响，而对各种脂肪因子对PCOS患者卵泡发育微环境影响研究较少。
Apelin为孤独G蛋白偶联受体-血管紧张素Ⅱ受体1相关的受体蛋白APJ的天然配体，apelin广泛分布于脑、胃、心、肺、肾、乳腺、子宫和卵巢等组织，参与血管平滑肌细胞、成骨细胞、血管内皮细胞等细胞增殖与分化调节。2005年研究发现鼠、人类皮下组织的脂肪细胞可分泌apelin，作为新发现的脂肪因子，诸多研究发现apelin参与IR相关的代谢综合征疾病，如肥胖、2型糖尿病、原发性高血压、PCOS等发病。有关apelin与PCOS发病研究中，王芳发现PCOS患者apelin水平显著高于对照者，apelin水平与BMI、腰围、Ins和HOMA-IR成正相关，BMI是影响血清apelin水平的独立相关因素；而G ren发现PCOS患者血清apelin水平升高与临床特征及生化指标之间无相关性；Chang发现PCOS患者血清apelin水平与对照组比较明显降低，并与载脂蛋白A负相关，可能参与远期心血管疾病发生。目前apelin与PCOS的研究处于初期阶段，但所有的研究表明，apelin可能参与PCOS发病，其具体作用机制有待研究。本研究通过检测PCOS患者血清apelin的表达，探讨其与患者脂、糖代谢和性激素之间的相关性；研究PCOS大鼠卵巢apelin/APJ表达情况和apelin对体外培养大鼠卵巢颗粒细胞影响，以进一步探索PCOS复杂的发病机制。本研究共分如下四部分：
第一部分：Apelin在多囊卵巢综合征患者血清中的表达及意义
目的检测PCOS患者血清apelin水平及与脂、糖代谢及性激素相关性。方法采用化学发光法检测50名PCOS患者及20名健康对照者空腹血糖（FBG）、胰岛素（Ins）、总胆固醇（TG）、甘油三酯（TC）、高密度脂蛋白（HDL）、低密度脂蛋白（LDL）水平；采取磁酶免疫法测定卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、孕酮（P）、催乳素（RPL）、睾酮（T）的血清浓度；采用酶联免疫法测受试验者血清apelin水平。结果（1）PCOS患者WHR[0.84（0.08)]、BMI[22.88（3.15）kg/m2]、T[0.66（0.35）ng/ml）、Ins[9.07（8.27）μu/ml]、LH[9.90（5.85）mIU/ml]、HOMA-IR[1.93(1.80)]与健康对照组WHR[0.79（0.05)]、BMI[（19.88（2.95）kg/m2]、T[0.29(0.23)ng/ml]、Ins[5.98(3.02)μu/ml]、LH[3.45(2.88)mIU/ml、HOMA-IR[1.26(1.43)]比较，差异均有统计学意义（P<0.05或P<0.001)。（2）PCOS患者血清apelin[501.40(98.66)pg/ml]明显高于对照者[416.77(61.07)pg/ml](P<0.001)；PCOS肥胖者apelin[520.80(100.27)pg/ml]、PCOS非肥胖者apelin[497.28(98.69)pg/ml]分别与对照组比较，差异均有统计学意义(P<0.001)；PCOS肥胖与非肥胖者apelin比较，差异有统计学意义（P<0.05）。（3）PCOS患者apelin水平与HOMA-IR、BMI、WHR、Ins、FBG成正相关(r=0.43，P=0.007；r=0.38，P=0.02；r=0.456，P=0.003；r=0.977，P=0.000；r=0.335，P=0.017)，与HDL(r=-0.456，P=0.005)成负相关。（4）二项logistic回归分析显示apelin与PCOS发病有关(P<0.05)。结论（1）PCOS患者血清apelin水平明显高于正常对照者。（2）PCOS患者血清apelin水平与肥胖及IR密切相关，apelin可能与PCOS发病有关。
第二部份：Apelin/APJ在PCOS大鼠卵巢中的表达及意义
目的探讨apelin/APJ在PCOS大鼠卵巢组织中的表达。方法21日龄SD鼠持续肌肉注射脱氢表雄酮（DHEA）21天，建立PCOS大鼠模型，观察体重、卵巢重量、HE染色观察卵巢改变；测定E2、T、P、FSH、LH水平；采用免疫组化法观察apelin/APJ在PCOS大鼠卵巢中分布与表达；qRT-PCR检测卵巢组织内apelin/APJ mRNA表达，W-B检测卵巢组织内apelin/APJ蛋白表达。结果（1）实验组大鼠体重(134.47±5.48)g与对照组（138.27±17.20)g比较，低于对照组，差异无统计学意义（P>0.05）；实验组大鼠卵巢重（79.87±16.34)g与对照组（91.13±21.95)g比较，低于对照组，差异无统计学意义（P>0.05）；实验组大鼠卵巢/体重（0.67±0.17）与对照组（0.57±0.06）比较，差异有统计学意义（P<0.05）。（2）实验组大鼠血清LH（2.80±1.67）mIU/ml、T （3.28±2.00)ng/ml，与对照组LH （0.55±0.24） mIU/ml、 T（0.31±0.20)ng/ml比较，差异均有统计学意义（P<0.001），实验组FSH（1.43±0.72）mIU/ml、P（2.54±0.86）pg/ml、E2（24.35±6.99）pg/ml与对照组FSH（1.03±0.33）mIU/ml、P（3.14±0.88）ng/ml、E（237.33±14.15）pg/ml比较，差异无统计学意义（P>0.05）。（3）实验组卵巢组织HE染色见卵巢内有较多的早期发育小卵泡和未及时闭锁的囊状大卵泡，少见黄体，形成典型的多囊样改变卵巢，对照组卵巢内有不同阶段卵泡发育，可见黄体，无明显扩张的囊状卵泡。（4）免疫组织化学染色发现apelin/APJ分别表达于大鼠卵泡膜细胞、颗粒细胞中；实验组卵泡apelin蛋白表达的平均灰度值（623.16±14.58），对照组卵泡apelin蛋白表达平均灰度值（166.19±10.06），apelin在实验组表达量明显高于对照组(P<0.05)；实验组APJ蛋白表达平均灰度值为（813.56±15.75），对照组APJ蛋白表达的平均灰度值（231.21±12.08），实验组APJ表达量明显高于对照组(P<0.05)。（5）实验组卵巢内apelin mRNA的相对表达量(4.61±1.93)，对照组的相对表达量为(1.04±0.36)，两组比较差异有统计学意义(P<0.05)；实验组APJ mRNA的表达量(6.81±0.94)，对照组相对表达量（1.25±0.38），两组比较，差异有统计学意义(P<0.05)。（5）实验组apelin蛋白相对表达量（0.52±0.13），对照组apelin蛋白相对表达量（0.31±0.04），两组比较，差异有统计学意义(P<0.05)；实验组APJ蛋白的相对表达(0.76±0.24)，对照组表达量为（0.37±0.15），两组比较差异有统计学意义(P<0.05)。结论（1）DHEA可成功诱导PCOS大鼠模型。（2）apelin/APJ在大鼠卵泡内膜细胞和颗粒细胞均为阳性表达。（3）PCOS大鼠卵巢组织apelin/APJ表达明显高于对照组，可能与PCOS局部卵泡异常发育的复杂病因有关。
第三部分：Apelin对大鼠卵巢颗粒细胞增殖和凋亡的影响
目的探讨apelin对大鼠卵巢颗粒细胞增殖和凋亡的影响。方法原代培养SD大鼠卵巢颗粒细胞，贴壁分组后，对照组加入等容量无血清DMEM/F12培养基，低浓度组加入等容量apelin10-8mol/l，高浓度组加入等容量apelin10-6mol/l，继续培养12h、24h、48h，采用MTT法及流式细胞技术（Annexin/PI）法检测颗粒细胞增殖和凋亡率。结果（1）颗粒细胞培育12h、24h、48h后采用MTT检测，低浓度组吸光值分别为（0.30±0.03）、（0.58±0.06）、（0.64±0.02）、高浓度组分别为（0.26±0.02）、（0.50±0.11）、（0.58±0.01），两组同一时间点分别与对照组（0.13±0.02）、（0.33±0.07）、（0.35±0.02）比较，差异均有统计学意义(P<0.05)；48h低浓度组吸光值明显高于对照组及高浓度组(P<0.001，P<0.05)。（2）Annexin/PI检测细胞12h、24h、48h凋亡率，低浓度组分别为（8.15±0.43）%、（7.16±0.86）%、（6.07±0.33）%，高浓度分别为（8.45±0.89）%、（9.67±0.27）%、（12.63±0.39）%，两组同一时间点分别与空白对照组（8.85±0.57）%、（10.24±0.25）%、（15.43±0.85）%比较，差异均有统计学意义（P<0.05）；在48h低浓度组细胞凋亡率均明显低于对照组及高浓度（P<0.001）。结论（1）高浓度和低浓度组apelin均可促进颗粒细胞增殖，48h低浓度apelin促大鼠卵巢颗粒细胞增殖作用最明显。（2）高浓度和低浓度组apelin均可抑制颗粒细胞凋亡，48h低浓度apelin抑制大鼠卵巢颗粒细胞凋亡作用最明显。
第四部分：Apelin对大鼠卵巢颗粒细胞作用信号的通路研究
目的观察apelin对大鼠卵巢颗粒细胞促增殖抑制凋亡作用的信号通路。方法（1）观察apelin10-8mol/l作用颗粒细胞0、5、15、30、60min后Akt及磷酸化Akt（pAkt）蛋白表达情况。（2）用小分子RNA干扰技术观察APJ-siRNA001、APJ-siRNA002、APJ-siRNA003在50nM、100nM浓度12h、24h、48h对颗粒细胞的转染率及对颗粒细胞APJ蛋白抑制情况；结果100nMAPJ-siRNA002转染颗粒细胞48h后可沉默APJ表达。（3）将培育的颗粒细胞分为对照组A.apelin10-8mol/l组，实验组B.apelin10-8mol/l+100nM APJ-siRNA002组，实验组C.apelin10-8mol/l+PI-3K信号抑制剂LY294002组，实验组D.apelin10-8mol/l+Akt信号抑制剂HIM0组，采用MTT观察4组细胞培育48h后增殖情况及检测Akt、p-Akt、Bad、Bax、Foxo3a、Bc1-2蛋白表达变化。结果（1）apelin10-8mol/l在5min开始激活信号Akt激酶蛋白，15min达高峰，APJ-siRNA、阻断剂LY294002和阻断剂HIM0可抑制Akt激酶磷酸化。（2）细胞培育48h后进行MTT检测A、B、C、D组细胞吸光值分别为(0.61±0.03)、（0.34±0.03)、（0.36±0.01)、（0.38±0.04)，B、C、D组与对照A组比较，差异均有统计学意义(P<0.05)。（3）B、C、D组促进Bad、Bax、Foxo3a蛋白表达上调，与A组比较，差异有统计学意义(P<0.05)；B、C、D组抑制Bc1-2蛋白表达下调，与A组比较，差异有统计学意义(P<0.05)。结论（1）apelin10-8mol/l可以激活Akt激酶；APJ-siRNA002、PI-3K信号阻断剂LY294002和Akt信号阻断剂HIM0均可抑制其激活。（2）apelin10-8mol/l促细胞增殖抗凋亡效应可被APJ-siRNA002、PI-3K信号阻断剂LY294002和Akt信号阻断剂HIM0抑制。（2）ape1in与APJ受体结合通过PI-3K/Akt信号通路参与大鼠卵巢颗粒细胞增殖与凋亡调节。
Polycystic ovary syndrome(PCOS) is the most commonendocrinopathy associated with metabolism dysfunction in women ofreproductive age，with the incidence4%-10%，which is a heterogeneousdisease characterized by oligo-ovulation or chronic anovulation，features ofhyperandrogenism and/or biochemical hyperandrogenism，insulin resistanceand polycystic ovarian morphology on ultrasonograph. Compared with thehealthy women， PCOS women with menstrual disorder， infertility，hirsutism and acne have a greater risk for cardiovascular disease， diabetesmellitus， tumorous.Women with PCOS has a serious negative impact onhealth-related quality of life， affecting both physical and psychologicalwell-being. Follicular developmental anomaly is the main cause ofoligomenorrhea or amenorrhea， infertility in reproductive age women withPCOS. The study about follicular developmental failure in PCOS can help usto understand the pathogenesis of PCOS. The numbers of primordialfollicles in ovaries of patients with PCOS and healthy women were similar，while the number of growing and atresia follicles were increased，and the number of over-recruitment follicles recruited into the growing phase fail tomature even result in the appearance of polycystic ovaries， ovulationfailure. Some scholar propose that PCOS is a primarily follicle disease. Inspite of the mechanism of abnormal folliculogenes is unclear， manyresearches showed that alteration of many factors may directly or indirectlyregulation the microenvironment of folliculogenes through endocrine andlocal paracrine/autocrine actions.
In anovulatory women with PCOS and obesity undergoing a lifestyleprogram， resume ovulation women lose more body weight and abdominalfat than remain anovulatory women. In addition， this study shows thatconsistent loss of intra-abdominal fat is associated with resumption ofovulation. It is indicated that adipokines correlated with obesity or insulinresistance play important roles in the development of follicles. Recently，some domestic and international researches focuses on various adipokinesgene expression and/or protein function defects in insulin resistance possiblemechanisms of patients with PCOS， while the relationship betweenadipokines and follicular microenvironment in patients with PCOS has notbeen reported yet. Apelin is an endogenous ligand of the orphanG-protein-coupled receptor APJ， which is consisting of77amino acidresidues and exists in multiple molecular forms. Apelin signaling pathwayplays a role in the central and peripheral regulation of the cardiovascularsystem， such as blood pressure， in water and food intake， and possibly in immune function. In2005， studies detected the apelin can secrete by ratand human hypodermic adipose cell. Many studies explained thepathogenesis between apelin and insulin resistance correlated disease，such as2type diabetes mellitus、primarily hypertension、PCOS. Wangstudied that insulin， HOMA-IR and apelin levels in women with PCOS，compared with healthy women；were significantly increased；apelin levelspositively correlated with BMI，WHR， insulin and HOMA-IR， and BMIwas the independent influencing factor for apelin levels changes. G renreported that the apelin level in women with PCOS were significantly highcompared with healthy group， while failed to found the relationshipbetween apelin levels and biochemical indicator in patient withPCOS.Increased apelin levels have been reported in women with PCOS，but not all previous studies.Chang reported that apelin levels were lowerthan that of healthy women and apelin was positively correlated withapolipoprotein A levels in patients with PCOS. All studies indicated that thechanges of apelin levels are associated with obesity and IR，and contribute，in part to pathogenesis of PCOS， while there is no study about themechanism of apelin in pathogenesis for women with PCOS. Our study wasdesigned to explore the apelin level in Chinese patients with PCOS and therole of apelin in lipometabolism，glycometabolism and insulin resistance；to study the apelin express in the rat ovary with PCOS and control group；to future exprole the effect apelin on the cultured rat granulose cells. The study including four chapters， as following：
Chapter1Serum apelin levels in women with Polycystic OvarySyndrome
[Abstract] Objective This study is designed in order to investigateserum apelin levels between PCOS patients and healthy subjects. MethodsThe basal levels of fasting blood glucose(FBG)， insulin （Ins），high-density lipoprotein(HDL)， low density lipoprotein(LDL)，triglycerides(TG)， total cholesterol(TC)， follicle-stimulating hormone(FSH)， luteinizing hormone (LH)， testosterone (T)，progestin(P)，prolactin(PRL)，estrogen(E2) and apelin were measured on fifty women withPCOS， and20healthy women. Subjects with PCOS were further dividedinto two subgroups according to body mass index (BMI) as BMI≥25kg/m2and BMI<25kg/m2.Results The apelin level of the PCOS groupwith[501.40(98.66)pg/ml]was higher than the controls[416.77(61.07)pg/ml](P<0.001).Both the BMI≥25kg/m2and BMI<25kg/m2women with PCOShad higher apelin levels，compared to the controls(P<0.001).Compared withthe BMI≥25kg/m2patients， the BMI<25kg/m2women with PCOS hadsignificantly lower apelin levels(P<0.05).A positive correlation was foundbetween apelin level and HOMA-IR、BMI and waist： hip ratio(WHR)、insulin、glucose in the patients with PCOS((r=0.43，P=0.007；r=0.38，P=0.02；r=0.456，P=0.003；r=0.977，P=0.000；r=0.335，P=0.017)，anda negative correlation was found between apelin and HDL(r=-0.456， P=0.005).Logistic analysis showed that apelin levels was significant in thedevelopment of PCOS(P<0.05).Conclusions Our study indicates that apelinin women with PCOS exhibit an increase along with obesity and IR. Furtherstudies are required to clarify the etiology and effects of apelin in womenwith PCOS.
Chapter2Expression of apelin/APJ in Polycystic Ovary Syndromeof rats
[Abstract] Objective To explore the role of apelin/APJ in thepathogenesis of PCOS.Methods Female SD rats aged21day were dividedinto two groups in pairs. The rats of experimental group were injected S.C.each day with dehydroeplandrosterone(DHEA) and the rats of control groupinjected with oil；Ovarian weight and sex hormone levels in rat blood of bothgroups were determined；Ovarian change was observed by HE staining. Theexpression of apelin and APJ were determined by immunohistochemical-staining in the ovaries； The levels of apelin and APJ mRNA and protein inovary tissue were measured by qRT-PCR and W-B technique. Results （1）The ovarian and body weight of model group were both lower than that ofthe control group（P>0.05），however， the ovarian weight/body weight ofmodel group was higher than that of control group（P<0.05）（.2）Comparedwith the control group the serum of LH and T in model group experiment group were significantly increased （P<0.05）.Although there was nosignificant difference of FSH，P and E2between the two groups（P>0.05）.（3）There were some early development of little follicles and theabundance of subcaplular ovarian cyst in ovary of model group， which wastypical polycystic ovary morphous，while some different development offollicles and corpus luteum were detected in ovary of control group，without subcaplular ovariancyst.(4)Immunostai-ning of apelin and APJ inthe granulosa cells and theca cells of PCOS rat’s ovaries were obviouslystronger than that of normal control (P<0.05， respectively).The levels ofapelin and APJ mRNA and protein in ovary of PCOS were obviously higherthan that of normal control (P<0.05， respectively).Conclusion Thedisruption of apelin/APJ system may be involved in the pathogenesis ofPCOS.
Chapter3Effects of apelin on proliferation and apoptosis of theovarian granulosa cells in SD rat
[Abstract] Objective To observe effect of apelin on rat ovariangranulosa cells proliferation and apoptosis in vitro. Method primary cultureof SD rat ovarian granulosa cells were cultured with DMEM/F12serum-freemedium without blood serum in control group， low concentration (apelin10-8mol/l) group， high concentration (apelin10-6mol/l) group， and tested by MTT assay and flow cytometry (Annexin/PI)method for the detection ofGC proliferation and apoptosis at12h、24h、48h.Results Apelin10-8mol/lcould significantly promote the proliferation of the GC at48h(P<0.05)andsignificantly inhibit apoptosis of the GC at48h(P<0.05).Conclusion Apelincould promote proliferation and suppress apoptosis of GC in vitro.
Chapter4Mechanism of apelin promote proliferation and supressapoptosis in granulosa cells
[Abstract] Objective T0investigate mechanism of apelin promoteproliferation and suppress apoptosis in GC. Method (1) The expression ofAkt and pAkt inGC cultured with apelin10-8mol/l at0、5、15、30、60minwas detected by W-B.(2)RNA interference was used to down-regulate theexpression of APJ in rat GC；Rat GC was divided into4groups：A.apelin10-8mol/l group，B.apelin10-8mol/l+APJ-siRNA.C.apelin10-8mol/l+PI-3k inhi--bitor LY294002group， D.apelin10-8mol/l+Akt inhibitor HIM0， theapoptosis rate of GC cultured after48h was detected by MTT；the proteinlevels of Akt、pAkt、Bad、Bax、Foxo3a and Bc1-2were detected by W-B.
Results (1) Apelin activated the phosphorylation of Akt kinase at5min，andthe peak at15min，however，the phosphorylation of Akt were blocked byAPJ-siRNA、inhibitor LY294002and HIMO.(2)The effect of apelin10-8mol/l promote proliferation were blocked by APJ-siRNA、 inhibitor LY294002and HIMO in GC (P<0.05，respectively).(3)Apelin10-8mol/lincreased Bcl-2protein， decreased bad、foxo3a and bax protein in GC，while，the effect were blocked by APJ-siRNA、inhibitor LY294002andHIMO(P<0.05，respectively).Conclusion These results indicate that theeffect of apelin promote proliferation and suppress apoptosis in GC ismediated via the APJ/PI-3K/Akt signaling pathway.
无排卵PCOS患者改变生活方式后，排卵恢复者的体重、腹部脂肪减少比未恢复者更明显；此外PCOS患者内脏脂肪持续减少与排卵恢复紧密相关。可见肥胖及IR相关脂肪因子在PCOS患者卵泡发育中有重要影响，近年大量国内外研究侧重于探讨脂肪因子如脂联素、瘦素等在PCOS患者基因表达和（或）蛋白功能缺陷对脂、糖代谢影响，而对各种脂肪因子对PCOS患者卵泡发育微环境影响研究较少。
Apelin为孤独G蛋白偶联受体-血管紧张素Ⅱ受体1相关的受体蛋白APJ的天然配体，apelin广泛分布于脑、胃、心、肺、肾、乳腺、子宫和卵巢等组织，参与血管平滑肌细胞、成骨细胞、血管内皮细胞等细胞增殖与分化调节。2005年研究发现鼠、人类皮下组织的脂肪细胞可分泌apelin，作为新发现的脂肪因子，诸多研究发现apelin参与IR相关的代谢综合征疾病，如肥胖、2型糖尿病、原发性高血压、PCOS等发病。有关apelin与PCOS发病研究中，王芳发现PCOS患者apelin水平显著高于对照者，apelin水平与BMI、腰围、Ins和HOMA-IR成正相关，BMI是影响血清apelin水平的独立相关因素；而G ren发现PCOS患者血清apelin水平升高与临床特征及生化指标之间无相关性；Chang发现PCOS患者血清apelin水平与对照组比较明显降低，并与载脂蛋白A负相关，可能参与远期心血管疾病发生。目前apelin与PCOS的研究处于初期阶段，但所有的研究表明，apelin可能参与PCOS发病，其具体作用机制有待研究。本研究通过检测PCOS患者血清apelin的表达，探讨其与患者脂、糖代谢和性激素之间的相关性；研究PCOS大鼠卵巢apelin/APJ表达情况和apelin对体外培养大鼠卵巢颗粒细胞影响，以进一步探索PCOS复杂的发病机制。本研究共分如下四部分：
第一部分：Apelin在多囊卵巢综合征患者血清中的表达及意义
目的检测PCOS患者血清apelin水平及与脂、糖代谢及性激素相关性。方法采用化学发光法检测50名PCOS患者及20名健康对照者空腹血糖（FBG）、胰岛素（Ins）、总胆固醇（TG）、甘油三酯（TC）、高密度脂蛋白（HDL）、低密度脂蛋白（LDL）水平；采取磁酶免疫法测定卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E2）、孕酮（P）、催乳素（RPL）、睾酮（T）的血清浓度；采用酶联免疫法测受试验者血清apelin水平。结果（1）PCOS患者WHR[0.84（0.08)]、BMI[22.88（3.15）kg/m2]、T[0.66（0.35）ng/ml）、Ins[9.07（8.27）μu/ml]、LH[9.90（5.85）mIU/ml]、HOMA-IR[1.93(1.80)]与健康对照组WHR[0.79（0.05)]、BMI[（19.88（2.95）kg/m2]、T[0.29(0.23)ng/ml]、Ins[5.98(3.02)μu/ml]、LH[3.45(2.88)mIU/ml、HOMA-IR[1.26(1.43)]比较，差异均有统计学意义（P<0.05或P<0.001)。（2）PCOS患者血清apelin[501.40(98.66)pg/ml]明显高于对照者[416.77(61.07)pg/ml](P<0.001)；PCOS肥胖者apelin[520.80(100.27)pg/ml]、PCOS非肥胖者apelin[497.28(98.69)pg/ml]分别与对照组比较，差异均有统计学意义(P<0.001)；PCOS肥胖与非肥胖者apelin比较，差异有统计学意义（P<0.05）。（3）PCOS患者apelin水平与HOMA-IR、BMI、WHR、Ins、FBG成正相关(r=0.43，P=0.007；r=0.38，P=0.02；r=0.456，P=0.003；r=0.977，P=0.000；r=0.335，P=0.017)，与HDL(r=-0.456，P=0.005)成负相关。（4）二项logistic回归分析显示apelin与PCOS发病有关(P<0.05)。结论（1）PCOS患者血清apelin水平明显高于正常对照者。（2）PCOS患者血清apelin水平与肥胖及IR密切相关，apelin可能与PCOS发病有关。
第二部份：Apelin/APJ在PCOS大鼠卵巢中的表达及意义
目的探讨apelin/APJ在PCOS大鼠卵巢组织中的表达。方法21日龄SD鼠持续肌肉注射脱氢表雄酮（DHEA）21天，建立PCOS大鼠模型，观察体重、卵巢重量、HE染色观察卵巢改变；测定E2、T、P、FSH、LH水平；采用免疫组化法观察apelin/APJ在PCOS大鼠卵巢中分布与表达；qRT-PCR检测卵巢组织内apelin/APJ mRNA表达，W-B检测卵巢组织内apelin/APJ蛋白表达。结果（1）实验组大鼠体重(134.47±5.48)g与对照组（138.27±17.20)g比较，低于对照组，差异无统计学意义（P>0.05）；实验组大鼠卵巢重（79.87±16.34)g与对照组（91.13±21.95)g比较，低于对照组，差异无统计学意义（P>0.05）；实验组大鼠卵巢/体重（0.67±0.17）与对照组（0.57±0.06）比较，差异有统计学意义（P<0.05）。（2）实验组大鼠血清LH（2.80±1.67）mIU/ml、T （3.28±2.00)ng/ml，与对照组LH （0.55±0.24） mIU/ml、 T（0.31±0.20)ng/ml比较，差异均有统计学意义（P<0.001），实验组FSH（1.43±0.72）mIU/ml、P（2.54±0.86）pg/ml、E2（24.35±6.99）pg/ml与对照组FSH（1.03±0.33）mIU/ml、P（3.14±0.88）ng/ml、E（237.33±14.15）pg/ml比较，差异无统计学意义（P>0.05）。（3）实验组卵巢组织HE染色见卵巢内有较多的早期发育小卵泡和未及时闭锁的囊状大卵泡，少见黄体，形成典型的多囊样改变卵巢，对照组卵巢内有不同阶段卵泡发育，可见黄体，无明显扩张的囊状卵泡。（4）免疫组织化学染色发现apelin/APJ分别表达于大鼠卵泡膜细胞、颗粒细胞中；实验组卵泡apelin蛋白表达的平均灰度值（623.16±14.58），对照组卵泡apelin蛋白表达平均灰度值（166.19±10.06），apelin在实验组表达量明显高于对照组(P<0.05)；实验组APJ蛋白表达平均灰度值为（813.56±15.75），对照组APJ蛋白表达的平均灰度值（231.21±12.08），实验组APJ表达量明显高于对照组(P<0.05)。（5）实验组卵巢内apelin mRNA的相对表达量(4.61±1.93)，对照组的相对表达量为(1.04±0.36)，两组比较差异有统计学意义(P<0.05)；实验组APJ mRNA的表达量(6.81±0.94)，对照组相对表达量（1.25±0.38），两组比较，差异有统计学意义(P<0.05)。（5）实验组apelin蛋白相对表达量（0.52±0.13），对照组apelin蛋白相对表达量（0.31±0.04），两组比较，差异有统计学意义(P<0.05)；实验组APJ蛋白的相对表达(0.76±0.24)，对照组表达量为（0.37±0.15），两组比较差异有统计学意义(P<0.05)。结论（1）DHEA可成功诱导PCOS大鼠模型。（2）apelin/APJ在大鼠卵泡内膜细胞和颗粒细胞均为阳性表达。（3）PCOS大鼠卵巢组织apelin/APJ表达明显高于对照组，可能与PCOS局部卵泡异常发育的复杂病因有关。
第三部分：Apelin对大鼠卵巢颗粒细胞增殖和凋亡的影响
目的探讨apelin对大鼠卵巢颗粒细胞增殖和凋亡的影响。方法原代培养SD大鼠卵巢颗粒细胞，贴壁分组后，对照组加入等容量无血清DMEM/F12培养基，低浓度组加入等容量apelin10-8mol/l，高浓度组加入等容量apelin10-6mol/l，继续培养12h、24h、48h，采用MTT法及流式细胞技术（Annexin/PI）法检测颗粒细胞增殖和凋亡率。结果（1）颗粒细胞培育12h、24h、48h后采用MTT检测，低浓度组吸光值分别为（0.30±0.03）、（0.58±0.06）、（0.64±0.02）、高浓度组分别为（0.26±0.02）、（0.50±0.11）、（0.58±0.01），两组同一时间点分别与对照组（0.13±0.02）、（0.33±0.07）、（0.35±0.02）比较，差异均有统计学意义(P<0.05)；48h低浓度组吸光值明显高于对照组及高浓度组(P<0.001，P<0.05)。（2）Annexin/PI检测细胞12h、24h、48h凋亡率，低浓度组分别为（8.15±0.43）%、（7.16±0.86）%、（6.07±0.33）%，高浓度分别为（8.45±0.89）%、（9.67±0.27）%、（12.63±0.39）%，两组同一时间点分别与空白对照组（8.85±0.57）%、（10.24±0.25）%、（15.43±0.85）%比较，差异均有统计学意义（P<0.05）；在48h低浓度组细胞凋亡率均明显低于对照组及高浓度（P<0.001）。结论（1）高浓度和低浓度组apelin均可促进颗粒细胞增殖，48h低浓度apelin促大鼠卵巢颗粒细胞增殖作用最明显。（2）高浓度和低浓度组apelin均可抑制颗粒细胞凋亡，48h低浓度apelin抑制大鼠卵巢颗粒细胞凋亡作用最明显。
第四部分：Apelin对大鼠卵巢颗粒细胞作用信号的通路研究
目的观察apelin对大鼠卵巢颗粒细胞促增殖抑制凋亡作用的信号通路。方法（1）观察apelin10-8mol/l作用颗粒细胞0、5、15、30、60min后Akt及磷酸化Akt（pAkt）蛋白表达情况。（2）用小分子RNA干扰技术观察APJ-siRNA001、APJ-siRNA002、APJ-siRNA003在50nM、100nM浓度12h、24h、48h对颗粒细胞的转染率及对颗粒细胞APJ蛋白抑制情况；结果100nMAPJ-siRNA002转染颗粒细胞48h后可沉默APJ表达。（3）将培育的颗粒细胞分为对照组A.apelin10-8mol/l组，实验组B.apelin10-8mol/l+100nM APJ-siRNA002组，实验组C.apelin10-8mol/l+PI-3K信号抑制剂LY294002组，实验组D.apelin10-8mol/l+Akt信号抑制剂HIM0组，采用MTT观察4组细胞培育48h后增殖情况及检测Akt、p-Akt、Bad、Bax、Foxo3a、Bc1-2蛋白表达变化。结果（1）apelin10-8mol/l在5min开始激活信号Akt激酶蛋白，15min达高峰，APJ-siRNA、阻断剂LY294002和阻断剂HIM0可抑制Akt激酶磷酸化。（2）细胞培育48h后进行MTT检测A、B、C、D组细胞吸光值分别为(0.61±0.03)、（0.34±0.03)、（0.36±0.01)、（0.38±0.04)，B、C、D组与对照A组比较，差异均有统计学意义(P<0.05)。（3）B、C、D组促进Bad、Bax、Foxo3a蛋白表达上调，与A组比较，差异有统计学意义(P<0.05)；B、C、D组抑制Bc1-2蛋白表达下调，与A组比较，差异有统计学意义(P<0.05)。结论（1）apelin10-8mol/l可以激活Akt激酶；APJ-siRNA002、PI-3K信号阻断剂LY294002和Akt信号阻断剂HIM0均可抑制其激活。（2）apelin10-8mol/l促细胞增殖抗凋亡效应可被APJ-siRNA002、PI-3K信号阻断剂LY294002和Akt信号阻断剂HIM0抑制。（2）ape1in与APJ受体结合通过PI-3K/Akt信号通路参与大鼠卵巢颗粒细胞增殖与凋亡调节。
Polycystic ovary syndrome(PCOS) is the most commonendocrinopathy associated with metabolism dysfunction in women ofreproductive age，with the incidence4%-10%，which is a heterogeneousdisease characterized by oligo-ovulation or chronic anovulation，features ofhyperandrogenism and/or biochemical hyperandrogenism，insulin resistanceand polycystic ovarian morphology on ultrasonograph. Compared with thehealthy women， PCOS women with menstrual disorder， infertility，hirsutism and acne have a greater risk for cardiovascular disease， diabetesmellitus， tumorous.Women with PCOS has a serious negative impact onhealth-related quality of life， affecting both physical and psychologicalwell-being. Follicular developmental anomaly is the main cause ofoligomenorrhea or amenorrhea， infertility in reproductive age women withPCOS. The study about follicular developmental failure in PCOS can help usto understand the pathogenesis of PCOS. The numbers of primordialfollicles in ovaries of patients with PCOS and healthy women were similar，while the number of growing and atresia follicles were increased，and the number of over-recruitment follicles recruited into the growing phase fail tomature even result in the appearance of polycystic ovaries， ovulationfailure. Some scholar propose that PCOS is a primarily follicle disease. Inspite of the mechanism of abnormal folliculogenes is unclear， manyresearches showed that alteration of many factors may directly or indirectlyregulation the microenvironment of folliculogenes through endocrine andlocal paracrine/autocrine actions.
In anovulatory women with PCOS and obesity undergoing a lifestyleprogram， resume ovulation women lose more body weight and abdominalfat than remain anovulatory women. In addition， this study shows thatconsistent loss of intra-abdominal fat is associated with resumption ofovulation. It is indicated that adipokines correlated with obesity or insulinresistance play important roles in the development of follicles. Recently，some domestic and international researches focuses on various adipokinesgene expression and/or protein function defects in insulin resistance possiblemechanisms of patients with PCOS， while the relationship betweenadipokines and follicular microenvironment in patients with PCOS has notbeen reported yet. Apelin is an endogenous ligand of the orphanG-protein-coupled receptor APJ， which is consisting of77amino acidresidues and exists in multiple molecular forms. Apelin signaling pathwayplays a role in the central and peripheral regulation of the cardiovascularsystem， such as blood pressure， in water and food intake， and possibly in immune function. In2005， studies detected the apelin can secrete by ratand human hypodermic adipose cell. Many studies explained thepathogenesis between apelin and insulin resistance correlated disease，such as2type diabetes mellitus、primarily hypertension、PCOS. Wangstudied that insulin， HOMA-IR and apelin levels in women with PCOS，compared with healthy women；were significantly increased；apelin levelspositively correlated with BMI，WHR， insulin and HOMA-IR， and BMIwas the independent influencing factor for apelin levels changes. G renreported that the apelin level in women with PCOS were significantly highcompared with healthy group， while failed to found the relationshipbetween apelin levels and biochemical indicator in patient withPCOS.Increased apelin levels have been reported in women with PCOS，but not all previous studies.Chang reported that apelin levels were lowerthan that of healthy women and apelin was positively correlated withapolipoprotein A levels in patients with PCOS. All studies indicated that thechanges of apelin levels are associated with obesity and IR，and contribute，in part to pathogenesis of PCOS， while there is no study about themechanism of apelin in pathogenesis for women with PCOS. Our study wasdesigned to explore the apelin level in Chinese patients with PCOS and therole of apelin in lipometabolism，glycometabolism and insulin resistance；to study the apelin express in the rat ovary with PCOS and control group；to future exprole the effect apelin on the cultured rat granulose cells. The study including four chapters， as following：
Chapter1Serum apelin levels in women with Polycystic OvarySyndrome
[Abstract] Objective This study is designed in order to investigateserum apelin levels between PCOS patients and healthy subjects. MethodsThe basal levels of fasting blood glucose(FBG)， insulin （Ins），high-density lipoprotein(HDL)， low density lipoprotein(LDL)，triglycerides(TG)， total cholesterol(TC)， follicle-stimulating hormone(FSH)， luteinizing hormone (LH)， testosterone (T)，progestin(P)，prolactin(PRL)，estrogen(E2) and apelin were measured on fifty women withPCOS， and20healthy women. Subjects with PCOS were further dividedinto two subgroups according to body mass index (BMI) as BMI≥25kg/m2and BMI<25kg/m2.Results The apelin level of the PCOS groupwith[501.40(98.66)pg/ml]was higher than the controls[416.77(61.07)pg/ml](P<0.001).Both the BMI≥25kg/m2and BMI<25kg/m2women with PCOShad higher apelin levels，compared to the controls(P<0.001).Compared withthe BMI≥25kg/m2patients， the BMI<25kg/m2women with PCOS hadsignificantly lower apelin levels(P<0.05).A positive correlation was foundbetween apelin level and HOMA-IR、BMI and waist： hip ratio(WHR)、insulin、glucose in the patients with PCOS((r=0.43，P=0.007；r=0.38，P=0.02；r=0.456，P=0.003；r=0.977，P=0.000；r=0.335，P=0.017)，anda negative correlation was found between apelin and HDL(r=-0.456， P=0.005).Logistic analysis showed that apelin levels was significant in thedevelopment of PCOS(P<0.05).Conclusions Our study indicates that apelinin women with PCOS exhibit an increase along with obesity and IR. Furtherstudies are required to clarify the etiology and effects of apelin in womenwith PCOS.
Chapter2Expression of apelin/APJ in Polycystic Ovary Syndromeof rats
[Abstract] Objective To explore the role of apelin/APJ in thepathogenesis of PCOS.Methods Female SD rats aged21day were dividedinto two groups in pairs. The rats of experimental group were injected S.C.each day with dehydroeplandrosterone(DHEA) and the rats of control groupinjected with oil；Ovarian weight and sex hormone levels in rat blood of bothgroups were determined；Ovarian change was observed by HE staining. Theexpression of apelin and APJ were determined by immunohistochemical-staining in the ovaries； The levels of apelin and APJ mRNA and protein inovary tissue were measured by qRT-PCR and W-B technique. Results （1）The ovarian and body weight of model group were both lower than that ofthe control group（P>0.05），however， the ovarian weight/body weight ofmodel group was higher than that of control group（P<0.05）（.2）Comparedwith the control group the serum of LH and T in model group experiment group were significantly increased （P<0.05）.Although there was nosignificant difference of FSH，P and E2between the two groups（P>0.05）.（3）There were some early development of little follicles and theabundance of subcaplular ovarian cyst in ovary of model group， which wastypical polycystic ovary morphous，while some different development offollicles and corpus luteum were detected in ovary of control group，without subcaplular ovariancyst.(4)Immunostai-ning of apelin and APJ inthe granulosa cells and theca cells of PCOS rat’s ovaries were obviouslystronger than that of normal control (P<0.05， respectively).The levels ofapelin and APJ mRNA and protein in ovary of PCOS were obviously higherthan that of normal control (P<0.05， respectively).Conclusion Thedisruption of apelin/APJ system may be involved in the pathogenesis ofPCOS.
Chapter3Effects of apelin on proliferation and apoptosis of theovarian granulosa cells in SD rat
[Abstract] Objective To observe effect of apelin on rat ovariangranulosa cells proliferation and apoptosis in vitro. Method primary cultureof SD rat ovarian granulosa cells were cultured with DMEM/F12serum-freemedium without blood serum in control group， low concentration (apelin10-8mol/l) group， high concentration (apelin10-6mol/l) group， and tested by MTT assay and flow cytometry (Annexin/PI)method for the detection ofGC proliferation and apoptosis at12h、24h、48h.Results Apelin10-8mol/lcould significantly promote the proliferation of the GC at48h(P<0.05)andsignificantly inhibit apoptosis of the GC at48h(P<0.05).Conclusion Apelincould promote proliferation and suppress apoptosis of GC in vitro.
Chapter4Mechanism of apelin promote proliferation and supressapoptosis in granulosa cells
[Abstract] Objective T0investigate mechanism of apelin promoteproliferation and suppress apoptosis in GC. Method (1) The expression ofAkt and pAkt inGC cultured with apelin10-8mol/l at0、5、15、30、60minwas detected by W-B.(2)RNA interference was used to down-regulate theexpression of APJ in rat GC；Rat GC was divided into4groups：A.apelin10-8mol/l group，B.apelin10-8mol/l+APJ-siRNA.C.apelin10-8mol/l+PI-3k inhi--bitor LY294002group， D.apelin10-8mol/l+Akt inhibitor HIM0， theapoptosis rate of GC cultured after48h was detected by MTT；the proteinlevels of Akt、pAkt、Bad、Bax、Foxo3a and Bc1-2were detected by W-B.
Results (1) Apelin activated the phosphorylation of Akt kinase at5min，andthe peak at15min，however，the phosphorylation of Akt were blocked byAPJ-siRNA、inhibitor LY294002and HIMO.(2)The effect of apelin10-8mol/l promote proliferation were blocked by APJ-siRNA、 inhibitor LY294002and HIMO in GC (P<0.05，respectively).(3)Apelin10-8mol/lincreased Bcl-2protein， decreased bad、foxo3a and bax protein in GC，while，the effect were blocked by APJ-siRNA、inhibitor LY294002andHIMO(P<0.05，respectively).Conclusion These results indicate that theeffect of apelin promote proliferation and suppress apoptosis in GC ismediated via the APJ/PI-3K/Akt signaling pathway.
引文
[1] Naveed Sattar.PCOS，insulin resistance and long-term risks for diabetes and vasculardisease[J].Br J Diabetes Vasc Dis，2009，9(1)：15–18
[2] Goodarzi MO，Dumesic DA，Chazenbalk G，et al.Polycystic ovary syndromeetiology，pathogenesis and diagnosis[J].Nat Rev Endocrinol，2011，7(4)：219-231
[3] Azziz R，Marin C，Hoq L，et al.Healthe care related economic burden of thepolycystic ovary syndrome during the reproductive life span[J].J Clin Metab，2005，90(8)：4650–4658
[4] Andrea JC， Gerald FW， Valerie B，et al.Cardiometabolic risk in polycystic ovarysyndrome： a comparison of different approaches to defining the metabolicsyndrome[J].Hum Reprod，2008，23(10)：2352-2358
[5] Coffey S，Bano G，Mason HD，et al.Health-related quality of life in women withpolycystic ovary syndrome： A comparison with the general population using thePolycystic Ovary Syndrome Questionnaire (PCOSQ) and the Short Form-36(SF-36)[J].Gynaecol Endocrinol，2006，22(2)：80–86
[6] Hughesden，P.E.Morphology and morphogenesis of the Stein-Leventhal Ovary andof So-called“Hyperthecosis”[J].Obstet Gynaecol Surv，1982，37(2)：59–77
[7] Webber LJ， Stubbs S. Stark J，et al.Formation and early development of follicles inthe polycystic ovary[J].Lancet，2003，362(9389)：1017–1021
[8]邵华，侯丽辉，昊效科.多囊卵巢综合征的卵泡发育异常[J].现代妇产进展.2006，15(1)：64-68
[9] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17-33
[10] Mattioli M，Barboni B，Bacci ML，et a1．Maturation of pig oocytes：observationson membrane potential[J].Biol Reprod，1990，43(2)：318-322
[11]李慧蓉，魏兆莲.多囊卵巢综合征卵巢颗粒细胞凋亡研究进展[J].国际生殖健康/计划生育杂志，2009，28(5)：331-335
[12] Webber U，Stubbs S，Stark J，et a1.Prolonged survival in culture of preantral folliclesform polycystic ovaries[J].J Clin Endoerinol Metab，2007，92(5)：1975-1978
[13]王秀秀，李威，吴效科.多囊卵巢综合征无排卵与卵泡发育异常[J].医学研究杂志，2009，(38)：13-17
[14] Hussein MR.Apoptosis in the ovary：molecular mechanisms[J].Hum Reprod Update，2005，11(2)：161-177
[15] Chen Xl， Ni R， Mo Yq，et al.Appropriate BMI levels for PCOS patients inSouthern China[J].Hum Reprod，2010，25(5)：1295-1302
[16] Robker RL，Akison LK，Bennett BD，et al.Norman.Obese women exhibit differencesin ovarian metabolites， hormones， and gene expression compared withmoderate-weight women[J].J Clin Endocrinol Metab，2009，94(5)：1533-1540
[17] Kuchenbecker WK， Groen H，Asselt SJ，et al.In women with polycystic ovarysyndrome and obesity，loss of intra-abdominal fat is associated with resumption ofovulation[J].Hum Reprod，2011，26(9)：2505-2512
[18] Kleinz MJ， Davenport AP.Emerging roles of apelin in biology andmedicine[J].Pharmacol Ther，2005，107(2)：198-211
[19]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[20] G ren K，Sa s z N，Noyan V，et al.Plasma Apelin levels in patients with polycysticovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[21] Chang CY，Tsai YC，Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[1] Boucher J，Masri B，Daviand D，et a1.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[2] Soriguer F， Garrido L， Garcia S，et a1.Apelin levels are increased in morbidlyobese subjects with type2diabetes mellitus[J].Obes Surg，2009，19(11)：1574-80
[3] Sonmez A，Celebi G，Erdem G，et a1.Plasma apelin and ADMA levels in patientswith essential hypertension[J].Clin Exp Hypertens，2010，32(3)：179-83
[4]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[5] G ren K， Sa s z N，Noyan V， et al.Plasma apelin levels in patients withpolycystic ovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[6] Chang CY，Tsai YC， Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[7] Rotterdam ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group.Revised2003consensus on diagnostic criteria and long term health risks related topolycystic ovary syndrome[J].Fertil Steril，2004，81(1)：19–20
[8] Bernasconi D，Del Monte PP，Meozzi M，et al. The impact of obesity on hormonalparameters in hirsute women [J].Metabolism，1996，45(1)：72-75
[9] Bjorntop P. Abdominal obesity and the metabolic syndrome [J].Ann Med，1992，24(6)：465-468
[10] Naveed S. PCOS，insulin resistance and long-term risks for diabetes and vasculardisease[J].Br J Diabetes Vasc Dis，2009，9(1)：15-18
[11] Moran LJ，Misso ML，Wild RA，et al. Impaired glucose tolerance， type2diabetesand metabolic syndrome in polycystic ovary syndrome： a systematic review andmeta-analysis[J].Hum Reprod，2010，16（4）：347-363
[12] Wozniak SE， Gee L L，Wachtel M S，et al.Adipose tissue： the new endocrineorgan?A review article [J].Dig Dis Sci，2009，54(9)：1847-1856
[13] Ishida J， Hashimoto T， Hashimoto Y， et al.Regulatory roles for APJ， aseven-transmembrane receptor related to AT1，in blood pressure in vivo[J].J BiolChem，2004，279(25)：26274–26279
[14] Hashimoto T，Kihara M，Ishida J，et al.Apelin stimulates myosin light chainphosphorylation in vascular smooth muscle cells[J].Arterioscler Thromb Vasc Biol，2006，26(6)：1267-1272
[15] Kasai A，Shintani N，Oda M，et al．Apelin is a novel an giogenic factorin retinalendothelial cells[J]．Biochem Biophys Res Commun，2004，325(2)：395-400
[16] Saint Geniez M，Masri B，Malecaze F，et al．Expression of the murine m sr/APJrecptor an d its ligan d apelin is upregulated during formation of the retinalvessels[J]．Mech Dev，2002，110(1-2)：183-186
[17] Orio F Jr，Palomba S，Spinelli L，et al.The cardiovascular risk of young womenwith polycystic ovary syndrome： an observational， analytical， prospectivecase-control study[J].J Clin Endocrinol Metab，2004，89(8)：3696–3701
[18] Andrea JC，Gerald FW，Valerie B，et al.Cardiometabolic risk in polycystic ovarysyndrome： a comparison of different approaches to defining the metabolicsyndrome[J].Hum Reprod，2008，23(10)：2352-2358
[18] Takashi S， Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Anil ReprodSci，2009，116(1-2)：28-37
[20] Stefanie S，Bernadette A，Dieter S，et a1.The expression of apelin and its receptorAPJ during different physiological stages in the bovine ovary[J].Int J Biol Sci，2009，5(4)：344-350
[1] Boucher J，Masri B，Daviand D，et a1.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[2] Soriguer F，Garrido L，Garcia S，et a1.Apelin levels are increased in morbidly obesesubjects with type2diabetes mellitus[J].Obes Surg，2009，19(11)：1574-1580
[3] Sonmez A，Celebi G，Erdem G，et a1.Plasma apelin and ADMA Levels in patientswith essential hypertension[J].Clin Exp Hypertens，2010，32(3)：179-183
[4] Dray C，Debard C，Jager J，et a1.Apelin and APJ regulation in adipose tissue andskeletal muscle of type2diabetic mice and humans[J].Am J Physiol EndocrinolMetab，2010，298(6)：1161-1169
[5] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17–33
[6]苗竹林，王自能，杨艳东，等.TGF-β1在PCOS卵巢间质纤维化形成中的作用[J].生殖与避孕，2008，28(3)：174-178
[7] Takashi S，Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Ani ReprodSci，2009，116(1-2)：28-37
[8] Stefanie S，Bernadette A，Dieter S，et a1.The expression of Apelin and its receptorAPJ during different physiological stages in the bovine ovary[J].Int J Biol Sci，2009，5(4)：344-350
[9] Wei L，Hou XH，Tatemoto K，et a1.Regulation of apelin mRNA expression byinsulin and glucocorticoids in mouse3T3-L1adipocytes[J].Regul Pept，2005，132(1-3)：27-32
[10] Xie H，Tang SY， Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[11]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[12] Liu CH， Tao S， Li F， et al.PI3K/Akt signaling transduction pathway is involvedin rat vascular smooth muscle cell proliferation induced by apelin-13[J].ActaBiochim Biophys Sin，2010，42(6)：396-402
[13] Cui RR， Mao DA， Yi L，et al.Apelin suppresses apoptosis of human vascularsmooth muscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[14] Kidoya H， Ueno M，Yamada Y，et al.Spatial and temporal role of the apelin/APJsystem in the caliber size regulation of blood vessels during angiogenesis[J]. EMBOJ，2008，27(3)：522-534
[15] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved in ratvascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[1] Xie H，Tang SY， Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[2]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[3] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved inrat vascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[4] Cui RR， Mao DA，Yi L，et al.Apelin suppresses apoptosis of human vascularsmooth muscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[5]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398
[6] Hussein MR.Apoptosis in the ovary：molecular mechanisms[J].Hum Reprod Update，2005，11(2)：161-177
[7] Mattioli M，Barboni B，Bacci ML，et a1.Maturation of pig oocytes：observations onmembrane potential[J].Biol Reprod，1990，43(2)：318-322
[8]吴德华，姜桢，郭克芳，等.Apelin对肺血管内皮细胞增殖和抗凋亡的作用[J].复旦学报(医学版)，2008，35(5)：789-791
[9] Christopher M C，Susan LD，Melanie KM，et a1.Apelin，the ligand for theendothelial G-protein coupled receptor，APJ，is a potent angiogenic factor requiredfor normal vascular development of the frog embryo[J].Dev Biol，2006，296(1)：177-189
[10] Leon speroff主编.临床妇科内分泌学与不孕[M].山东科学技术出版社，2003，p372-372
[1] Xie H，Tang SY，Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[2]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[3] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved inrat vascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[4] Cui RR，Mao DA，Yi L，et al.Apelin suppresses apoptosis of human vascular smoothmuscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[5] Atsushi K，Yuki I，Toshihiko K，et al.Apelin Is a Crucial Factor for Hypoxia-InducedRetinal Angiogenesis[J].Arterioscler Thromb Vasc Biol，2010，30(11)：2182-2187
[6]张攀，沈伟.多囊卵巢综合征与PI-3K/Akt信号通路的关系[J].青岛农业大学学报(自然科学版)，2010，27(1)：11-16
[7] Fernanda P，Dalhia A，Griselda I，et al.Angiopoietin1reduces rat follicular atresiamediated by apoptosis through the PI3K/Akt pathway[J].Molecul Cellul Endocrin，2011，343(1-2)：79-87
[8]杨晓葵，宋蕊，贾婵维，等.大鼠窦状卵泡颗粒细胞中磷酸化蛋白激酶B表达与凋亡的关系[J].中国优生与遗传杂志，2010，18(8)：14-17
[9] Testa JR，Tsichlis PN.AKT signaling in normal and malignant cells[J].Oncog ene，2005，24(50)：7391–7393
[10]周华.胰岛素对大鼠血管平滑肌细胞PANKL表达影响及信号转导机制的研究
[D].湖南：湘雅二医院代谢内分泌学，2010
[11] Jonathan GJ，Coffer PJ，Ham J.Foxo transciption factors directly activate bim geneexpression and promote apoptosis in sympathetic neuron[J].J Cell Biol，2003，162(4)：613-622
[12] Dijkers PF，Medema RH，Lammers JW，et al.Expression of the pro-apoptotic Bcl-2family member Bim is regulated by the forkhead transcription factorFKHR-L1[J].Curr Biol，2000，10(19)：1201-1204
[13]孟春花.香猪卵巢细胞生物学特性和影响母猪卵巢颗粒细胞体外成熟的因素探讨[D].南京：南京农业大学动物遗传育种与繁殖，2008
[14] Hosaka T，Biggs W，Tieu D，et al.Disruption of forkhead transcription factor (FOXO)family members in mice reveals their functional diversification[J].PANS，2004，101(9)：2975-2980
[15] Kaipia A，Hsu SY，Hsueh AW.Expression and function of a proapoptotic Bcl-2family member Bcl-XL/Bcl-2-associated death promoter (Bad)in rat ovary[J]. Endo--crinology，1997，138(12)：5497-5505
[16] Hu XM，Christian P，Glenn IS.Expression and redistribution of cellular Bad，Bax，and Bcl-xL protein is associated with VCD-induced ovotoxicity in rats[J].BiolReprod，2001，65(5)1489-1495
[17] Shimojo N，Jesmin S，Zaedi S，et a1.Changes in important apoptosis-relatedmolecules in the endothelin-1-induced hypertrophied cardiomyocytes：effect of thepretreatment with eicosapentaenoic acid[J].Exp Biol Med，2006，231(6)：932-936
[18] Dorn GW.Apoptotic and non-apoptotic programmed cardiomyocyte death inventficular remodelling[J].Cardiovasc Re，2009，81(3)：465-473
[1] Naveed Sattar.PCOS，insulin resistance and long-term risks for diabetes and vasculardisease.Br J Diabetes Vasc Dis[J].2009，9(1)：15–18
[2] GoodarziMO，Dumesic DA，Chazenbalk G，et al.Polycystic ovary syndrome：etiology，pathogenesis and diagnosis[J].Nat Rev Endocrinol，2011，7(4)：219-231
[3] Hughesden PE.Morphology and morphogenesis of the Stein-Leventhal Ovary and ofso-called “Hyperthecosis”[J].Obstet and Gynaecol Surv，1982，37(2)：59-77
[4] Webber LJ， Stubbs S. Stark J，et al.Formation and early development of follicles inthe polycystic ovary[J].Lancet，2003，362(9389)：1017-1021
[5]邵华，侯丽辉，昊效科.多囊卵巢综合征的卵泡发育异常[J].现代妇产进展，2006，15(1)：64-68
[6]吴效科.多囊卵巢综合症可能是原发性卵泡病[J].中国实用妇科与产科杂志，2007，9，23(9)：660-663
[7] Artini PG，Monteleone P，Toldin MRP，et al.Growth factors and folliculogenesisin polycystic ovary patients[J].Expert Rev Endocrinol Metab，2007，2(2)：215–223
[8] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17–33
[9] Dumesic DA， Padmanabhan V，Abbott DH.Polycystic ovary syndrome and oocytedevelopmental competence[J].Obst Gyn Surv，2008，63(1)：39-48
[10] Dumesic DA，Abbott DH， Padmanabhan V.Polycystic ovary syndrome and itsdevelopmental origins[J].Rev Endocr Metab Discord，2007；8(2)：127-141
[11] Wood JR，Dumesic DA，Abbott DH，et al.Molecular abnormalities in oocytes fromwomen with polycystic ovary syndrome revealed by microarray analysis[J].J ClinEndocrinol Metab，2007，92(2)：705-713
[12] Pangas SA.Growth factors in ovarian development[J].Semin Reprod Med，2007，25(4)：225-34
[13] Miao ZL，Wang ZN，Yang YD，et al.Role of TGF-β1in the Formation of OvarianInterstitial Fibrosis in PCOS rat[J].J Reprod Contracep，2008，19(2)：83-92
[14] Chao KC，Ho CH，Shyong WY，et al.Anti-Mullerian hormone serum level as apredictive marker of ovarian function in Taiwanese women[J].J Chin Med Assoc，2012，75(2)：70-74
[15] Hart R，Doherty DA，Robert J，et al.Serum antimullerian hormone (AMH)levels areelevated in adolescent girls with polycystic ovaries and the polycystic ovariansyndrome(PCOS)[J]. Fertil Steril，2010，94(3)：1117-1121
[16] Almudena VL，DVM，Wen Y，et al.Developmental programming：prenataltestosterone excess disrupts anti-Mullerian hormone expression in preantral andantral follicles[J].Fertil Steril，2012，97(3)：748-756
[17] Patel SS，Carr BR.Oocyte quality in adult polycystic ovary syndrome[J]. SeminReprod Med2008，26(2)：196-203
[18] Franks S，Stark J，Hardy K.Follicle dynamics and anovulation in polycystic ovarysyndrome[J]. Hum Reprod Update，2008，14(4)：367-378
[19] Das M， Gillott DJ， Saridoan E，et al.Anti-Mullerian hormone is increased infollicular fluid from unstimulated ovaries in women with polycystic ovarysyndrome[J].Hum Reprod，2008，23(9)：2122-2126
[20] Dewailly D，Gronier H， Poncelet E，et al.Diagnosis of polycystic ovary syndrome(PCOS)： revisiting the threshold values of follicle count on ultrasound and of theserum AMH level for the definition of polycystic ovaries[J].Hum Reprod，2011，26，(11)：3123-3129
[21] Sasseville M，Ritter LJ，Nguyen TM，Liu F，et al.Growth differentiation factor9signaling requires ERK1/2activity in mouse granulosa and cumulus cells[J].J CellSci，2010，123(18)：3166-3176
[22] Hreinsson JG，Scott JE，Rasmussen C， et al.Growth differentiation factor-9promotes the growth，development， and survival of human ovarian follicles inorgan culture[J].J Clin Endocrinol Metab，2002，87(1)：316-21
[23] Zhao SY，Qiao J，Chen YJ，et al.Li.Expressionof growth differentiation factor-9andbone orphogenetic protein-15in oocytes and cumulus ranulosa cells of patients withpolycystic ovary yndrome[J].Fertil Steril，2010，94(1)：261-268
[24] Hsieh M， Zamah AM， Conti M. Epidermal growth factor-like growth factors in thefollicular fluid： role in oocyte development and maturation[J].Semin Reprod Med，2009，27(1)：52-61
[25] Jamnongjit M， Gill A，Hammes SR.Epidermal growth factor receptor signaling isrequired for normal ovarian steroidogenesis and oocyte maturation[J].Proc NatlAcad Sci USA，2005，102(45)：16257-16262
[26] Lin JX， Jia YD， Zhang CQ， et al.Effect of epidermal growth factor onfollicle-stimulating hormone-induced proliferation of granulosa cells from chickenprehierarchical follicles[J].J Zhejiang Univ Sci B，2011，12(11)：875-883
[27] Rusovici R，Hui YY，Lavoie HA.Epidermal growth factor-mediated inhibition offollicle-stimulating hormone-stimulated StAR gene expression in porcine granulosacells is associated with reduced histone H3acetylation[J].Biol Reprod，2005，72(4)：862-871
[28] Volpe A，Coukos G，D’Ambrogio G，et al.Follicular fluid steroid and epidermalgrowth factor content， and in vitro estrogen release by granulosaluteal cells frompatients with polycystic ovaries in an IVF/ET program[J].Eur J Obstet GynecolReprod Biol，1991；42(3)：195–199
[29] Violeta G，Katarina V，Saraga M，et al.Apoptosis and active caspase-3expression inhuman granulose cells[J].Fertil Steril，2005，83(2)：426-431
[30]张维怡，黄光英，刘洁，等.针刺对多囊卵巢综合征大鼠卵巢TGF-α、EGFR表达的影响[J].微循环学杂志，2008，18(4)：4-7
[31]王针织，俞超芹，蔡勤华，等.多囊卵巢综合征卵巢局部调控因子对卵泡生长发育的影响[J].中国妇产科临床杂志，2009，10(6)：471-475
[32] Marie PJ.Fibroblast growth factor signaling controlling bone formation： Anupdate.[J].Gene，2012，498(1)：2012-2021
[33] Bajram B，Fred S，Dieter S.Expression and localization of fibroblast growthfactor(FGF)family members during the final growth of bovine ovarianfollicles[J].Mol reprod dev，2004，67(2)：162–171
[34] Quennell JH， Stanton JAL，Hurst PR.Basic fibroblast growth factor expression inisolated small human ovarian follicles[J].Mol Hum Reprod，2004，10(9)：623–628
[35] Garor R，Abir R， Erman A，et al.Effects of basic fibroblast growth factor on in vitrodevelopment of human ovarian primordial follicles[J].Fertil Steril，2009，91(5)：1967-1975
[36] Gorar S，Culha C，Uc ZA，et al.Serum fibroblast growth factor21levels in polycysticovary syndrome[J].Gynecol Endocrinol，2010，26(11)：819-26
[37]潘洪娟，王继东，罗爽，等.FGF21在PCOS患者血清中的表达及意义[J].实用妇产科杂志，2011，27(8)：71-75
[38] Roeiy A，Chen X，Roberts VJ，et al.Expression of insulin-like growth factor-I (IGF-I)and IGF-II and the IGF-I，IGF-II，and insulin receptor genes and localization of thegene products in the human ovary[J].J clinical endocrinol metab，1993，77(5)：1411-8
[39] Fumio Otsuka， Shin Yamamoto，et al.Bone morphogenetic protein-15inhibitsfollicle-stimulating hormone (FSH) action by suppressing FSH receptorexpression[J].J biological chemistry，2001，276(14)：11387-11392
[40] Giudice LC.Growth factor action on ovarian function in polycysticovarysyndrome[J].Endocrinol Metab Clin North Am，1999，28(2)：325-339
[41] Rajaram S，Baylink DJ，Mohan S.Insulin-like growth factor-binding proteins in serumand other biological fluids：regulation and functions[J].Endocr Rev，1997，18(6)：801-31
[42] Sharma GT，Dubey PK，Kumar GS.Effects of IGF-1，TGF-α plus TGF-β1and bFGFon in vitro survival，growth and apoptosis in FSH-stimulated buffalo(Bubalisbubalus) preantral follicles[J].Growth Horm IGF Res，2010，20(4)：319-325
[43] Thomas FH，Campbell BK，Armstrong DG，et al.Effects of IGF-I bioavailability onbovine preantral follicular development in vitro[J].Reproduction，2007，133(6)：1121-1128
[44]徐淑云，赵淑萍，马德花，等.PCOS病人血清TNF-α及IGF-1水平变化及意义[J].青岛大学学报.2010，46(1)：46-48
[45] Wang H，Kwon MD，Choi.DH，et al.amRNA expression pattern of insulin-likegrowth factor components of granulosa cells and cumulus cells in women with andwithout polycystic ovary syndrome according to oocyte maturity[J].Fertil Steril，2010，6(94)：2417-2421
[46] Peng X，Maruo T，Samoto T，et al.Comparison of immunocytologic localization ofinsulin-like growth factor binding protein-4in normal and polycystic ovarysyndrome human ovaries[J].Endocr J，1996，43(3)：269-78
[47] Kwintkiewicz J，Nishi Y，Yanase T，et al. Peroxisome proliferatoractivated receptor-γmediates bisphenol A inhibition of FSH-stimulated IGF-1，aromatase，and estradiolin human granulosa cells[J].Environ Health Perspect，2010，118(3)：400–406
[48] Salas C，Julio-Pieper M，Valladares M，et al.Nerve growth factor-dependentactivation of trkA receptors in the human ovary results in synthesis of FSH receptorsand estrogen secretion[J].J Clin Endocrinol Metab，2006，91(6)：2396–2403
[49] Dissen GA， Cecilia GR，Sergio RO.Role of neurotrophic factors in early ovariandevelopment[J]. Semin Reprod Med，2009，27(1)：24-31
[50] Dissen GA，Cecilia GR，Paredes A，et al.Excessive ovarian production of nervegrowth factor facilitates development of cystic ovarian morphology in mice and is afeature of Polycystic Ovarian Syndrome in humans[J].Endocrinology，2009，150(6)：2906-2914
[51] Manneras L，Cajander S，L nn M，et al.Acupuncture and exercise restore adiposetissue expression of sympathetic markers and improve ovarian morphology in ratswith dihydrotestosterone-induced PCOS[J]. Am J Physiol Regul Integr CompPhysiol，2009，296(4)：1124-1131
[52] Blerkom J V，Antczak M，Schrander R.The developmental potential of the humanoocyte is related to the dissolved oxygen content of follicular fluid：association withvascular endothelial growth factor levels and perifollicular blood flowcharacteristics[J].Hum Reprod，1997，12（5）：1047-1055
[53] Agrawal R，Jacobs H，Panyne N，et al.Concentration of vascular endothelialgrowthfactor released by cultured human luteinized granulosecells is higher in women withpolycystic ovaries than in women withnormal ovaries[J].Fertil Steril，2002，78(6)：1164-1169
[54] Luo H，Kimura K，Aoki M，et al.Effect of vascular endothelial growth factor onmaturation，fertilization and developmental competence of bovine oocytes[J].J VetMed Sci，2002，64(9)：803-806
[55] Franks S，Robberts R，Hardy K，et al.Gonadotrophin regimens and oocyte quality inwomen with polycystic ovaries[J].Reprod Biomed Online，2003，6(2)：181–184
[56] Artini PG，Monti M，Matteucci C，et al.Vascular endothelial growth factor and basicfibroblast growth factor in polycystic ovary syndrome during controlled ovarianhyperstimulation[J].Gynecol Endocrinol，2006，22(8)：465-470
[57] Artini PG， Ruggiero M， Parisen Toldin MR，et al.Vascular endothelial growthfactor and its soluble receptor in patients with polycystic ovary syndromeundergoing IVF[J].Hum Fertil(Camb)，2009，12(1)：40-44
[58] Peitsidis P，Agrawal R.Role of vascular endothelial growth factor in women withPCO and PCOS：a systematic review[J].Reprod Biomed Online，2010，20(4)：444-452
[59]孙林，魏巍关，关咏梅.u-PA、U-PAR和PAI-1在睾酮联合hCG致多囊卵巢大鼠中的表达与意义[J].生殖与避孕，2006，26（10）：579-583
[60]申景欣，潘琼，薛敏，等.纤溶酶原激活因子及抑制因子在多囊卵巢综合征患者卵巢中的表达[J].第二军医大学学报，2010，12（2）：1309-1313
[61] Lindholm1A，Bixo M.Tissue plasminogen activator and plasminogen activatorinhibitor1in obese and lean patients with polycystic ovary syndrome[J]. GynecolEndocrinol，2010，26(10)：743-748
[62] Koiou E，Tziomalos K，Dinas K，et al.Plasma plasminogen activator inhibitor-1levelsin the different phenotypes of the polycystic ovary syndrome[J].Endocr J，2012，59(1)：21-9
[63] Lin YS，Tsai SJ，Lin MW，et al.Interleukin-6as an early chronic inflammatorymarker in Polycystic Ovary Syndrome with insulin receptor substrate-2polymorphism[J].Ame J Reprod mmunol，2011，66(6)：527-533
[64] Kim CH，Ahn JW，You RM，et al.Pioglitazone treatment decreases follicular fluidlevels of tumor necrosis factor-α and interleukin-6in patients with polycystic ovarysyndrome[J].Clin Exp Reprod Med.2011，38(2)：98–102
[65]游力，芦珂，赵跃然，等.血清抵抗素、C-反应蛋白及IL-6与PCOS的相关性研究[J].生殖与避孕，2010，30(5)：311-315
[66] Van Der Spuy ZM，Dyer SJ.The pathogenesis of infertility and early pregnancy lossin polycystic ovary syndrome[J].Best Pract Res Clin Obstet Gynaecol，2004，18(5)：755–771
[67]王秀霞，王天任，易晓芳，等.卵泡液中TNF-α和IFN-γ与卵丘颗粒细胞凋亡关系的探讨[J].生殖与避孕，2010，30(8)：519-522
[68]洪岭，滕晓明，李昆明，等.TNF-α在体外具有促进猪卵泡内膜细胞增殖的作用[J].华西医学，201025(1)：57-59
[69] Xiong YL，Liang XY，Yang X，et al.Low-grade chronic inflammation in theperipheral blood and ovaries of women with polycystic ovarian syndrome[J].Eur JObstet Gynecol Reprod Biol，2011，159(1)：148-150
[70]徐淑云，赵淑萍，马德花，等.PCOS病人血清TNF-α及IGF-1水平变化及意义[J].青岛大学医学院学报，2010，46(1)：45-48
[71] Pehlivanov B，Mitkov M.Serum leptin levels correlate with clinical and biochemicalindices of insulin resistance in women with polycystic ovary syndrome[J]，Eur JContracept Reprod Health Care，2009，14(2)：153-159
[72] Georgios A， Eleni K，Ioannis S，Vassilios L，et al.Serum and follicular fluid leptinlevels are correlated with human embryo quality[J].Reproduction，2005，130（1）：917–921
[73] Karamouti M，Kollia P，KallitsarisA，et al.Modulating effect of leptin on basal andfollicle stimulating hormone stimulated steroidogenesis in cultured human luteingranulosa cells[J].J Endocrinol，2009，32（5）：415-419
[74] Yildizhan R，Ilhan GA，Yildizhan B，et al.Serum retinol-binding protein4，leptin，and plasma asymmetric dimethylarginine levels in obese and nonobese youngwomen with polycystic ovary syndrome[J].Fertil Steril，2011，96(1)：246-251
[75] Seow KM，Juan CC，Hsu YP，Ho LT，Wang YY，Hwang JL.Serum and follicularresistin levels in women with polycystic ovarian syndrome during IVF-stimulatedcycles[J].Hum Reprod，2005，20(1)：117–121
[76]陆秀娥.多囊卵巢综合征患者颗粒细胞AQP9、RESISTIN表达及对体外受精结局的影响[D].浙江：浙江大学医学妇产科学生殖内分泌，2006
[77] Shen CJ，Tsai，EM，Lee JN，et al.The cocentrations of visfatin in the follicular fluidsof women undergoing controlled ovarian stimulation are correlated to the number ofoocytes retrieved[J].Fertil Steril，2010，93(6)：1844-1850
[78] Tan BK，Chen J，Digby JE，et al.Increased visfatin messenger ribonucleic acid andprotein levels in adipose tissue and adipocytes in women with polycystic ovarysyndrome：parallel increase in plasma visfatin[J].J Clin Endocrinol Metab，2006，91(12)：5022-5028
[79] Jongwutiwes T，Lertvikool S，Leelaphiwat S，et al.Serum visfatin in Asian womenwith polycystic ovary syndrome[J].Gynecol Endocrinol，2009，25(8)：536-542
[80] Chabrolle C，Tosca L，Ramé C，et al. Adiponectin increases insulin-like growthfactor-induced progesterone and estradiol secretion in human granulosacells[J].Fertil Steril，2009，92(6)：1988-1996
[81] Chappaz E，Albornoz MS，Campos D，et a1．Adiponectin enhances in vitrodevelopment of swine embryos[J].Domest Anjm Endocrin，2008，35(2)：198-207
[82] Louise MH，Leonhardt H，Kullberg J，et al.Adipose tissue has aberrant morphologyand function in PCOS：enlarged adipocytes and low serum adiponectin，but notcirculating sex steroids，are strongly associated with insulin resistance[J].J ClinEndocrinol Metab，2011，96(2)：304-311
[83]洪青青.多囊卵巢综合征患者子宫内膜和卵巢颗粒细胞脂联素及其受体表达的研究[D].上海，上海交通大学妇产科学，2009
[84] Matthias J，Kleinz，Anthony P，et al.Emerging roles of apelin in biology andmedicine[J].Pharmacology Therapeutics，2005，107(4)：198-211
[85]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[86] G ren K，Sa s z N，Noyan V，et al.Plasma apelin levels in patiets with polycysticovary syndrome [J]. J Turkish German Gynecol Assoc，2012，13(1)：27-31
[87] Chang CY，Tsai YC，Lee CH， et al.Lower serum apelin levels in women withpolycystic ovary syndrome [J].Fertil Steril，2011，95(8)：2520-2523
[88]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398
[89] Azziz R，Woods KS，Reyna R， et al. The prevalence and features of the polycysticovary syndrome in an unselected population [J].J Clin Endocrinol Metab，2004，89(6)：2745–2749
[90] Pasquali R.Obesity and androgens： facts and perspectives [J]. Fertil Steril，2006，85(5)：1319–1340
[91] Kuchenbecker WK， Groen H，Asselt SJ，et al.In women with polycystic ovarysyndrome and obesity， loss of intra-abdominal fat is associated with resumption ofovulation[J].Human Reprod.2011，26(9)：2505-2512
[1] Tatemoto K，Hosoya M，Habata Y，et al.Isolation and characterization of a novelendogenous peptide ligand for the human APJ receptor[J].Biochem Biophys ResCommun，1998，25(1)：471–476
[2] Takashi S，Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Ani ReprodSci，2009，116(1-2)：28-37
[3]丁永刚，倪安民.脂肪因子apelin与代谢综合征[J].中国糖尿病杂志，2008，16(7)：444-447
[4] De Falco M，De Luca L，Onori N，et al.Apelin expression in normal humantissues[J].In Vivo，2002，16(5)：333-336
[5] Kawamata Y， Habata Y，Fukusumi S，et al.Molecular properties of apelin：tissuedistribution and receptor binding[J].Biochim Biophys Acta，2001，1538(2-3)：162-171
[6] Habata Y，Fujii R， Hosoya M.Apelin， the natural ligand of the orphan receptorAPJ，is abundantly secreted in the colostrum[J].Biochim Biophys Acta，1999，1452(1)：25-35
[7] De Falco M，Fedele V，Russo T，et al.Distribution of apelin，the endogenous ligandof the APJ receptor，in the lizard Podarcis sicula[J].J Mol Histol，2004，35(5)：521-527
[8] Katugampola SD，Maguire JJ，Matthewson SR，et al.[(125)I]-(Pyr(1))Apelin-13isa novel radioligand for localizing the APJ orphan receptor in human and rat tissueswith evidence for a vasoconstrictor role in man[J].Br J Pharmacol，2001，132(6)：1255–1260
[9] Reaux A， De Mota N， Skultetyova I，et al.Physiological role of a novelneuropeptide，apelin，and its receptor in the rat brain[J].J Neurochem，2001，77(4)：1085–1096
[10] Choe H， Farzan M， Konkel M， et al. The orphan seven-transmembrane receptorapj supports the entry of primary T-cell-line-tropic and dualtropic humanimmunodeficiency virus type1[J].J Virol，1998，72(7)：6113–6118
[11]巴宏军，陈红珊，杜敏联，马华梅，等.肥胖女童血清apelin、chemerin水平及其与胰岛素抵抗的关系[J].中国病理生理杂志，2011，27(10)：2014-2016
[12] Katarzyna Z，Oswiecimska J，Swietochowska E，et al.Assessment of serum apehnlevels in girls with anorexia nervosa[J]．J Clin Endocfinol Metab，2010，95(6)：2935-3941
[13] Isabelle CL， Michaela V， Daviaud D，et al.Effect of hypocaloric diet-inducedweight loss in obese women on plasma apelin and adipose tissue expression ofapelin and APJ[J].Eur J Endocrinol，2008158(6)：905-910
[14] Boucher J，Masri B， Daviaud D，et al.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[15] Kunduzova O， Alet1N， Delesque TN，et al.Apelin/APJ signaling system： apotential link between adipose tissue and endothelial angiogenic processes[J]. FAS--EB J，2008，(22）：4146-4154
[16] Li L，Yang G，Li Q．Changes and relations of circulating visfatin，apelin，and resistinlevels in normal，impaired glucose tolerance，and type2diabetic subjects[J]. ExpClin Endocrinol Diabet，2006，114(10)：544-548
[17]杨冬莲，黎英荣，连莲，等.肥胖2型糖尿病患者血清apelin的变化及其与血脂的关系研究[J].广西医科大学学报，2010，27(4)：565-568
[18] Dray C，Debard C，Jager J，et al.Apelin and APJ regulation in adipose tissue andskeletal muscle of type2diabetic mice and humans[J].Am J Physiol EndocrinolMetab，2010，298(6)：1161-1169
[19] Erdem G，Dogru T，Tasci I，et al.Apelin levels in newly diagnosed type2diabetesmellitus[J].Exp Clin Endocrinol Diabetes，2008，116(5)：289-292
[20] Zhang Y，Shen C，Li X，et al.Low plasma apelin in newly diagnosed type2diabetesin Chinese people[J].Diabetes Care，2009，32(12)：150-150
[21] Kadoglou N PE，Tsanikidis H，Kapelouzou A，et al.Effects of rosiglitazone andmetformin treatment on apelin，visfatin，and ghrelin levels in patients with type2diabetes mellitus[J].Metabolism，2010，59(3)：373-379
[22] Trayhurn P，Beattie JH．Physiological role of adipose tissue：white adipose tissue asan endocrine and secretory organ[J].Proc Nutr Soc，2001，60(3)：329-339
[23] Higuchi K，Masaki T，Gotoh K，et a1.Apelin，an APJ receptor ligand，regulates bodyadiposity and favors the mRNA expression of uncoupling proteinsi nmice[J].Endo--crinology，2007，148(6)：2690-2697
[24]李庆辉，杨天伦，谭碧峰.原发性高血压患者血清apelin-13与脂联素的相关性研究[J].医学临床研究，2011，28(2)：223-226
[25] Strazynska A，Bryl W，Hoffmann K，et al.The estimation of concentartion of apelin，insulin， fasting glucose and anthropometric factors in young hypertensivepatients[J].J Hypertens，2010，28(28)：187-187
[26] Monika PK，Tomasz K，Andrzej M，et al.Reduced circulating apelin in essentialhypertension and its association with cardiac dysfunction[J].J Hypertens，2011，29(5)：971-979
[27]廖予妹，乔福元．妊娠期高血压疾病患者胎盘组织中血管活性肽apelin的表达及其意义[J].中华妇产科杂志，2007，42(6)：382-386
[28] Ishida J，Hashimoto J，Hashimoto Y，et a1．Regulatory roles for APJ，a seventransmembrane receptor related to angiotensin type I receptor in blood pressure invivo[J]．Bio1Chem，2004，279(25)：26274-26279
[29] Hashinoto T，Kihara M，Ishida I，et a1．Apelin Stimulates myosin light chainphosphory1ation in vascular smooth muscle cells[J]．Arterioscler Thromb VascBiol，2006，26(6)：1267-1272
[30]李丽英，魏雪梅，王伟，等.Apelin/APJ系统研究新进展[J].重庆医学，2011，40（22）：2275-2279
[31]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[32] G ren K，Sa s z N，Noyan V，et al.Plasma Apelin levels in patiets with polycysticovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[33] Chang CY， Tsai YC，Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[34]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398
[2] Goodarzi MO，Dumesic DA，Chazenbalk G，et al.Polycystic ovary syndromeetiology，pathogenesis and diagnosis[J].Nat Rev Endocrinol，2011，7(4)：219-231
[3] Azziz R，Marin C，Hoq L，et al.Healthe care related economic burden of thepolycystic ovary syndrome during the reproductive life span[J].J Clin Metab，2005，90(8)：4650–4658
[4] Andrea JC， Gerald FW， Valerie B，et al.Cardiometabolic risk in polycystic ovarysyndrome： a comparison of different approaches to defining the metabolicsyndrome[J].Hum Reprod，2008，23(10)：2352-2358
[5] Coffey S，Bano G，Mason HD，et al.Health-related quality of life in women withpolycystic ovary syndrome： A comparison with the general population using thePolycystic Ovary Syndrome Questionnaire (PCOSQ) and the Short Form-36(SF-36)[J].Gynaecol Endocrinol，2006，22(2)：80–86
[6] Hughesden，P.E.Morphology and morphogenesis of the Stein-Leventhal Ovary andof So-called“Hyperthecosis”[J].Obstet Gynaecol Surv，1982，37(2)：59–77
[7] Webber LJ， Stubbs S. Stark J，et al.Formation and early development of follicles inthe polycystic ovary[J].Lancet，2003，362(9389)：1017–1021
[8]邵华，侯丽辉，昊效科.多囊卵巢综合征的卵泡发育异常[J].现代妇产进展.2006，15(1)：64-68
[9] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17-33
[10] Mattioli M，Barboni B，Bacci ML，et a1．Maturation of pig oocytes：observationson membrane potential[J].Biol Reprod，1990，43(2)：318-322
[11]李慧蓉，魏兆莲.多囊卵巢综合征卵巢颗粒细胞凋亡研究进展[J].国际生殖健康/计划生育杂志，2009，28(5)：331-335
[12] Webber U，Stubbs S，Stark J，et a1.Prolonged survival in culture of preantral folliclesform polycystic ovaries[J].J Clin Endoerinol Metab，2007，92(5)：1975-1978
[13]王秀秀，李威，吴效科.多囊卵巢综合征无排卵与卵泡发育异常[J].医学研究杂志，2009，(38)：13-17
[14] Hussein MR.Apoptosis in the ovary：molecular mechanisms[J].Hum Reprod Update，2005，11(2)：161-177
[15] Chen Xl， Ni R， Mo Yq，et al.Appropriate BMI levels for PCOS patients inSouthern China[J].Hum Reprod，2010，25(5)：1295-1302
[16] Robker RL，Akison LK，Bennett BD，et al.Norman.Obese women exhibit differencesin ovarian metabolites， hormones， and gene expression compared withmoderate-weight women[J].J Clin Endocrinol Metab，2009，94(5)：1533-1540
[17] Kuchenbecker WK， Groen H，Asselt SJ，et al.In women with polycystic ovarysyndrome and obesity，loss of intra-abdominal fat is associated with resumption ofovulation[J].Hum Reprod，2011，26(9)：2505-2512
[18] Kleinz MJ， Davenport AP.Emerging roles of apelin in biology andmedicine[J].Pharmacol Ther，2005，107(2)：198-211
[19]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[20] G ren K，Sa s z N，Noyan V，et al.Plasma Apelin levels in patients with polycysticovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[21] Chang CY，Tsai YC，Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[1] Boucher J，Masri B，Daviand D，et a1.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[2] Soriguer F， Garrido L， Garcia S，et a1.Apelin levels are increased in morbidlyobese subjects with type2diabetes mellitus[J].Obes Surg，2009，19(11)：1574-80
[3] Sonmez A，Celebi G，Erdem G，et a1.Plasma apelin and ADMA levels in patientswith essential hypertension[J].Clin Exp Hypertens，2010，32(3)：179-83
[4]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[5] G ren K， Sa s z N，Noyan V， et al.Plasma apelin levels in patients withpolycystic ovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[6] Chang CY，Tsai YC， Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[7] Rotterdam ESHRE/ASRM-Sponsored PCOS Consensus Workshop Group.Revised2003consensus on diagnostic criteria and long term health risks related topolycystic ovary syndrome[J].Fertil Steril，2004，81(1)：19–20
[8] Bernasconi D，Del Monte PP，Meozzi M，et al. The impact of obesity on hormonalparameters in hirsute women [J].Metabolism，1996，45(1)：72-75
[9] Bjorntop P. Abdominal obesity and the metabolic syndrome [J].Ann Med，1992，24(6)：465-468
[10] Naveed S. PCOS，insulin resistance and long-term risks for diabetes and vasculardisease[J].Br J Diabetes Vasc Dis，2009，9(1)：15-18
[11] Moran LJ，Misso ML，Wild RA，et al. Impaired glucose tolerance， type2diabetesand metabolic syndrome in polycystic ovary syndrome： a systematic review andmeta-analysis[J].Hum Reprod，2010，16（4）：347-363
[12] Wozniak SE， Gee L L，Wachtel M S，et al.Adipose tissue： the new endocrineorgan?A review article [J].Dig Dis Sci，2009，54(9)：1847-1856
[13] Ishida J， Hashimoto T， Hashimoto Y， et al.Regulatory roles for APJ， aseven-transmembrane receptor related to AT1，in blood pressure in vivo[J].J BiolChem，2004，279(25)：26274–26279
[14] Hashimoto T，Kihara M，Ishida J，et al.Apelin stimulates myosin light chainphosphorylation in vascular smooth muscle cells[J].Arterioscler Thromb Vasc Biol，2006，26(6)：1267-1272
[15] Kasai A，Shintani N，Oda M，et al．Apelin is a novel an giogenic factorin retinalendothelial cells[J]．Biochem Biophys Res Commun，2004，325(2)：395-400
[16] Saint Geniez M，Masri B，Malecaze F，et al．Expression of the murine m sr/APJrecptor an d its ligan d apelin is upregulated during formation of the retinalvessels[J]．Mech Dev，2002，110(1-2)：183-186
[17] Orio F Jr，Palomba S，Spinelli L，et al.The cardiovascular risk of young womenwith polycystic ovary syndrome： an observational， analytical， prospectivecase-control study[J].J Clin Endocrinol Metab，2004，89(8)：3696–3701
[18] Andrea JC，Gerald FW，Valerie B，et al.Cardiometabolic risk in polycystic ovarysyndrome： a comparison of different approaches to defining the metabolicsyndrome[J].Hum Reprod，2008，23(10)：2352-2358
[18] Takashi S， Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Anil ReprodSci，2009，116(1-2)：28-37
[20] Stefanie S，Bernadette A，Dieter S，et a1.The expression of apelin and its receptorAPJ during different physiological stages in the bovine ovary[J].Int J Biol Sci，2009，5(4)：344-350
[1] Boucher J，Masri B，Daviand D，et a1.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[2] Soriguer F，Garrido L，Garcia S，et a1.Apelin levels are increased in morbidly obesesubjects with type2diabetes mellitus[J].Obes Surg，2009，19(11)：1574-1580
[3] Sonmez A，Celebi G，Erdem G，et a1.Plasma apelin and ADMA Levels in patientswith essential hypertension[J].Clin Exp Hypertens，2010，32(3)：179-183
[4] Dray C，Debard C，Jager J，et a1.Apelin and APJ regulation in adipose tissue andskeletal muscle of type2diabetic mice and humans[J].Am J Physiol EndocrinolMetab，2010，298(6)：1161-1169
[5] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17–33
[6]苗竹林，王自能，杨艳东，等.TGF-β1在PCOS卵巢间质纤维化形成中的作用[J].生殖与避孕，2008，28(3)：174-178
[7] Takashi S，Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Ani ReprodSci，2009，116(1-2)：28-37
[8] Stefanie S，Bernadette A，Dieter S，et a1.The expression of Apelin and its receptorAPJ during different physiological stages in the bovine ovary[J].Int J Biol Sci，2009，5(4)：344-350
[9] Wei L，Hou XH，Tatemoto K，et a1.Regulation of apelin mRNA expression byinsulin and glucocorticoids in mouse3T3-L1adipocytes[J].Regul Pept，2005，132(1-3)：27-32
[10] Xie H，Tang SY， Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[11]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[12] Liu CH， Tao S， Li F， et al.PI3K/Akt signaling transduction pathway is involvedin rat vascular smooth muscle cell proliferation induced by apelin-13[J].ActaBiochim Biophys Sin，2010，42(6)：396-402
[13] Cui RR， Mao DA， Yi L，et al.Apelin suppresses apoptosis of human vascularsmooth muscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[14] Kidoya H， Ueno M，Yamada Y，et al.Spatial and temporal role of the apelin/APJsystem in the caliber size regulation of blood vessels during angiogenesis[J]. EMBOJ，2008，27(3)：522-534
[15] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved in ratvascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[1] Xie H，Tang SY， Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[2]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[3] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved inrat vascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[4] Cui RR， Mao DA，Yi L，et al.Apelin suppresses apoptosis of human vascularsmooth muscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[5]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398
[6] Hussein MR.Apoptosis in the ovary：molecular mechanisms[J].Hum Reprod Update，2005，11(2)：161-177
[7] Mattioli M，Barboni B，Bacci ML，et a1.Maturation of pig oocytes：observations onmembrane potential[J].Biol Reprod，1990，43(2)：318-322
[8]吴德华，姜桢，郭克芳，等.Apelin对肺血管内皮细胞增殖和抗凋亡的作用[J].复旦学报(医学版)，2008，35(5)：789-791
[9] Christopher M C，Susan LD，Melanie KM，et a1.Apelin，the ligand for theendothelial G-protein coupled receptor，APJ，is a potent angiogenic factor requiredfor normal vascular development of the frog embryo[J].Dev Biol，2006，296(1)：177-189
[10] Leon speroff主编.临床妇科内分泌学与不孕[M].山东科学技术出版社，2003，p372-372
[1] Xie H，Tang SY，Cui RR，et al.Apelin and its receptor are expressed in humanosteoblasts[J].Regul Pept，2006，134(2-3)：118-125
[2]谢辉，李辉，梁秋华，等.Apelin抑制人成骨细胞的凋亡[J].中华骨质疏松和骨矿盐疾病杂志，2008，1(l)：39-44
[3] Liu CH，Tao S，Li F，et al.PI3K/Akt signaling transduction pathway is involved inrat vascular smooth muscle cell proliferation induced by apelin-13[J].Acta BiochimBiophys Sin，2010，42(6)：396-402
[4] Cui RR，Mao DA，Yi L，et al.Apelin suppresses apoptosis of human vascular smoothmuscle cells via APJ/PI3-K/Akt signaling pathways[J].Amino Acids，2010，39(5)：1193–1200
[5] Atsushi K，Yuki I，Toshihiko K，et al.Apelin Is a Crucial Factor for Hypoxia-InducedRetinal Angiogenesis[J].Arterioscler Thromb Vasc Biol，2010，30(11)：2182-2187
[6]张攀，沈伟.多囊卵巢综合征与PI-3K/Akt信号通路的关系[J].青岛农业大学学报(自然科学版)，2010，27(1)：11-16
[7] Fernanda P，Dalhia A，Griselda I，et al.Angiopoietin1reduces rat follicular atresiamediated by apoptosis through the PI3K/Akt pathway[J].Molecul Cellul Endocrin，2011，343(1-2)：79-87
[8]杨晓葵，宋蕊，贾婵维，等.大鼠窦状卵泡颗粒细胞中磷酸化蛋白激酶B表达与凋亡的关系[J].中国优生与遗传杂志，2010，18(8)：14-17
[9] Testa JR，Tsichlis PN.AKT signaling in normal and malignant cells[J].Oncog ene，2005，24(50)：7391–7393
[10]周华.胰岛素对大鼠血管平滑肌细胞PANKL表达影响及信号转导机制的研究
[D].湖南：湘雅二医院代谢内分泌学，2010
[11] Jonathan GJ，Coffer PJ，Ham J.Foxo transciption factors directly activate bim geneexpression and promote apoptosis in sympathetic neuron[J].J Cell Biol，2003，162(4)：613-622
[12] Dijkers PF，Medema RH，Lammers JW，et al.Expression of the pro-apoptotic Bcl-2family member Bim is regulated by the forkhead transcription factorFKHR-L1[J].Curr Biol，2000，10(19)：1201-1204
[13]孟春花.香猪卵巢细胞生物学特性和影响母猪卵巢颗粒细胞体外成熟的因素探讨[D].南京：南京农业大学动物遗传育种与繁殖，2008
[14] Hosaka T，Biggs W，Tieu D，et al.Disruption of forkhead transcription factor (FOXO)family members in mice reveals their functional diversification[J].PANS，2004，101(9)：2975-2980
[15] Kaipia A，Hsu SY，Hsueh AW.Expression and function of a proapoptotic Bcl-2family member Bcl-XL/Bcl-2-associated death promoter (Bad)in rat ovary[J]. Endo--crinology，1997，138(12)：5497-5505
[16] Hu XM，Christian P，Glenn IS.Expression and redistribution of cellular Bad，Bax，and Bcl-xL protein is associated with VCD-induced ovotoxicity in rats[J].BiolReprod，2001，65(5)1489-1495
[17] Shimojo N，Jesmin S，Zaedi S，et a1.Changes in important apoptosis-relatedmolecules in the endothelin-1-induced hypertrophied cardiomyocytes：effect of thepretreatment with eicosapentaenoic acid[J].Exp Biol Med，2006，231(6)：932-936
[18] Dorn GW.Apoptotic and non-apoptotic programmed cardiomyocyte death inventficular remodelling[J].Cardiovasc Re，2009，81(3)：465-473
[1] Naveed Sattar.PCOS，insulin resistance and long-term risks for diabetes and vasculardisease.Br J Diabetes Vasc Dis[J].2009，9(1)：15–18
[2] GoodarziMO，Dumesic DA，Chazenbalk G，et al.Polycystic ovary syndrome：etiology，pathogenesis and diagnosis[J].Nat Rev Endocrinol，2011，7(4)：219-231
[3] Hughesden PE.Morphology and morphogenesis of the Stein-Leventhal Ovary and ofso-called “Hyperthecosis”[J].Obstet and Gynaecol Surv，1982，37(2)：59-77
[4] Webber LJ， Stubbs S. Stark J，et al.Formation and early development of follicles inthe polycystic ovary[J].Lancet，2003，362(9389)：1017-1021
[5]邵华，侯丽辉，昊效科.多囊卵巢综合征的卵泡发育异常[J].现代妇产进展，2006，15(1)：64-68
[6]吴效科.多囊卵巢综合症可能是原发性卵泡病[J].中国实用妇科与产科杂志，2007，9，23(9)：660-663
[7] Artini PG，Monteleone P，Toldin MRP，et al.Growth factors and folliculogenesisin polycystic ovary patients[J].Expert Rev Endocrinol Metab，2007，2(2)：215–223
[8] Qiao J，Feng HL.Extra-and intra-ovarian factors in polycystic ovary syndrome：impact on oocyte maturation and embryo developmental competence[J].HumReprod Update，2011，17(1)：17–33
[9] Dumesic DA， Padmanabhan V，Abbott DH.Polycystic ovary syndrome and oocytedevelopmental competence[J].Obst Gyn Surv，2008，63(1)：39-48
[10] Dumesic DA，Abbott DH， Padmanabhan V.Polycystic ovary syndrome and itsdevelopmental origins[J].Rev Endocr Metab Discord，2007；8(2)：127-141
[11] Wood JR，Dumesic DA，Abbott DH，et al.Molecular abnormalities in oocytes fromwomen with polycystic ovary syndrome revealed by microarray analysis[J].J ClinEndocrinol Metab，2007，92(2)：705-713
[12] Pangas SA.Growth factors in ovarian development[J].Semin Reprod Med，2007，25(4)：225-34
[13] Miao ZL，Wang ZN，Yang YD，et al.Role of TGF-β1in the Formation of OvarianInterstitial Fibrosis in PCOS rat[J].J Reprod Contracep，2008，19(2)：83-92
[14] Chao KC，Ho CH，Shyong WY，et al.Anti-Mullerian hormone serum level as apredictive marker of ovarian function in Taiwanese women[J].J Chin Med Assoc，2012，75(2)：70-74
[15] Hart R，Doherty DA，Robert J，et al.Serum antimullerian hormone (AMH)levels areelevated in adolescent girls with polycystic ovaries and the polycystic ovariansyndrome(PCOS)[J]. Fertil Steril，2010，94(3)：1117-1121
[16] Almudena VL，DVM，Wen Y，et al.Developmental programming：prenataltestosterone excess disrupts anti-Mullerian hormone expression in preantral andantral follicles[J].Fertil Steril，2012，97(3)：748-756
[17] Patel SS，Carr BR.Oocyte quality in adult polycystic ovary syndrome[J]. SeminReprod Med2008，26(2)：196-203
[18] Franks S，Stark J，Hardy K.Follicle dynamics and anovulation in polycystic ovarysyndrome[J]. Hum Reprod Update，2008，14(4)：367-378
[19] Das M， Gillott DJ， Saridoan E，et al.Anti-Mullerian hormone is increased infollicular fluid from unstimulated ovaries in women with polycystic ovarysyndrome[J].Hum Reprod，2008，23(9)：2122-2126
[20] Dewailly D，Gronier H， Poncelet E，et al.Diagnosis of polycystic ovary syndrome(PCOS)： revisiting the threshold values of follicle count on ultrasound and of theserum AMH level for the definition of polycystic ovaries[J].Hum Reprod，2011，26，(11)：3123-3129
[21] Sasseville M，Ritter LJ，Nguyen TM，Liu F，et al.Growth differentiation factor9signaling requires ERK1/2activity in mouse granulosa and cumulus cells[J].J CellSci，2010，123(18)：3166-3176
[22] Hreinsson JG，Scott JE，Rasmussen C， et al.Growth differentiation factor-9promotes the growth，development， and survival of human ovarian follicles inorgan culture[J].J Clin Endocrinol Metab，2002，87(1)：316-21
[23] Zhao SY，Qiao J，Chen YJ，et al.Li.Expressionof growth differentiation factor-9andbone orphogenetic protein-15in oocytes and cumulus ranulosa cells of patients withpolycystic ovary yndrome[J].Fertil Steril，2010，94(1)：261-268
[24] Hsieh M， Zamah AM， Conti M. Epidermal growth factor-like growth factors in thefollicular fluid： role in oocyte development and maturation[J].Semin Reprod Med，2009，27(1)：52-61
[25] Jamnongjit M， Gill A，Hammes SR.Epidermal growth factor receptor signaling isrequired for normal ovarian steroidogenesis and oocyte maturation[J].Proc NatlAcad Sci USA，2005，102(45)：16257-16262
[26] Lin JX， Jia YD， Zhang CQ， et al.Effect of epidermal growth factor onfollicle-stimulating hormone-induced proliferation of granulosa cells from chickenprehierarchical follicles[J].J Zhejiang Univ Sci B，2011，12(11)：875-883
[27] Rusovici R，Hui YY，Lavoie HA.Epidermal growth factor-mediated inhibition offollicle-stimulating hormone-stimulated StAR gene expression in porcine granulosacells is associated with reduced histone H3acetylation[J].Biol Reprod，2005，72(4)：862-871
[28] Volpe A，Coukos G，D’Ambrogio G，et al.Follicular fluid steroid and epidermalgrowth factor content， and in vitro estrogen release by granulosaluteal cells frompatients with polycystic ovaries in an IVF/ET program[J].Eur J Obstet GynecolReprod Biol，1991；42(3)：195–199
[29] Violeta G，Katarina V，Saraga M，et al.Apoptosis and active caspase-3expression inhuman granulose cells[J].Fertil Steril，2005，83(2)：426-431
[30]张维怡，黄光英，刘洁，等.针刺对多囊卵巢综合征大鼠卵巢TGF-α、EGFR表达的影响[J].微循环学杂志，2008，18(4)：4-7
[31]王针织，俞超芹，蔡勤华，等.多囊卵巢综合征卵巢局部调控因子对卵泡生长发育的影响[J].中国妇产科临床杂志，2009，10(6)：471-475
[32] Marie PJ.Fibroblast growth factor signaling controlling bone formation： Anupdate.[J].Gene，2012，498(1)：2012-2021
[33] Bajram B，Fred S，Dieter S.Expression and localization of fibroblast growthfactor(FGF)family members during the final growth of bovine ovarianfollicles[J].Mol reprod dev，2004，67(2)：162–171
[34] Quennell JH， Stanton JAL，Hurst PR.Basic fibroblast growth factor expression inisolated small human ovarian follicles[J].Mol Hum Reprod，2004，10(9)：623–628
[35] Garor R，Abir R， Erman A，et al.Effects of basic fibroblast growth factor on in vitrodevelopment of human ovarian primordial follicles[J].Fertil Steril，2009，91(5)：1967-1975
[36] Gorar S，Culha C，Uc ZA，et al.Serum fibroblast growth factor21levels in polycysticovary syndrome[J].Gynecol Endocrinol，2010，26(11)：819-26
[37]潘洪娟，王继东，罗爽，等.FGF21在PCOS患者血清中的表达及意义[J].实用妇产科杂志，2011，27(8)：71-75
[38] Roeiy A，Chen X，Roberts VJ，et al.Expression of insulin-like growth factor-I (IGF-I)and IGF-II and the IGF-I，IGF-II，and insulin receptor genes and localization of thegene products in the human ovary[J].J clinical endocrinol metab，1993，77(5)：1411-8
[39] Fumio Otsuka， Shin Yamamoto，et al.Bone morphogenetic protein-15inhibitsfollicle-stimulating hormone (FSH) action by suppressing FSH receptorexpression[J].J biological chemistry，2001，276(14)：11387-11392
[40] Giudice LC.Growth factor action on ovarian function in polycysticovarysyndrome[J].Endocrinol Metab Clin North Am，1999，28(2)：325-339
[41] Rajaram S，Baylink DJ，Mohan S.Insulin-like growth factor-binding proteins in serumand other biological fluids：regulation and functions[J].Endocr Rev，1997，18(6)：801-31
[42] Sharma GT，Dubey PK，Kumar GS.Effects of IGF-1，TGF-α plus TGF-β1and bFGFon in vitro survival，growth and apoptosis in FSH-stimulated buffalo(Bubalisbubalus) preantral follicles[J].Growth Horm IGF Res，2010，20(4)：319-325
[43] Thomas FH，Campbell BK，Armstrong DG，et al.Effects of IGF-I bioavailability onbovine preantral follicular development in vitro[J].Reproduction，2007，133(6)：1121-1128
[44]徐淑云，赵淑萍，马德花，等.PCOS病人血清TNF-α及IGF-1水平变化及意义[J].青岛大学学报.2010，46(1)：46-48
[45] Wang H，Kwon MD，Choi.DH，et al.amRNA expression pattern of insulin-likegrowth factor components of granulosa cells and cumulus cells in women with andwithout polycystic ovary syndrome according to oocyte maturity[J].Fertil Steril，2010，6(94)：2417-2421
[46] Peng X，Maruo T，Samoto T，et al.Comparison of immunocytologic localization ofinsulin-like growth factor binding protein-4in normal and polycystic ovarysyndrome human ovaries[J].Endocr J，1996，43(3)：269-78
[47] Kwintkiewicz J，Nishi Y，Yanase T，et al. Peroxisome proliferatoractivated receptor-γmediates bisphenol A inhibition of FSH-stimulated IGF-1，aromatase，and estradiolin human granulosa cells[J].Environ Health Perspect，2010，118(3)：400–406
[48] Salas C，Julio-Pieper M，Valladares M，et al.Nerve growth factor-dependentactivation of trkA receptors in the human ovary results in synthesis of FSH receptorsand estrogen secretion[J].J Clin Endocrinol Metab，2006，91(6)：2396–2403
[49] Dissen GA， Cecilia GR，Sergio RO.Role of neurotrophic factors in early ovariandevelopment[J]. Semin Reprod Med，2009，27(1)：24-31
[50] Dissen GA，Cecilia GR，Paredes A，et al.Excessive ovarian production of nervegrowth factor facilitates development of cystic ovarian morphology in mice and is afeature of Polycystic Ovarian Syndrome in humans[J].Endocrinology，2009，150(6)：2906-2914
[51] Manneras L，Cajander S，L nn M，et al.Acupuncture and exercise restore adiposetissue expression of sympathetic markers and improve ovarian morphology in ratswith dihydrotestosterone-induced PCOS[J]. Am J Physiol Regul Integr CompPhysiol，2009，296(4)：1124-1131
[52] Blerkom J V，Antczak M，Schrander R.The developmental potential of the humanoocyte is related to the dissolved oxygen content of follicular fluid：association withvascular endothelial growth factor levels and perifollicular blood flowcharacteristics[J].Hum Reprod，1997，12（5）：1047-1055
[53] Agrawal R，Jacobs H，Panyne N，et al.Concentration of vascular endothelialgrowthfactor released by cultured human luteinized granulosecells is higher in women withpolycystic ovaries than in women withnormal ovaries[J].Fertil Steril，2002，78(6)：1164-1169
[54] Luo H，Kimura K，Aoki M，et al.Effect of vascular endothelial growth factor onmaturation，fertilization and developmental competence of bovine oocytes[J].J VetMed Sci，2002，64(9)：803-806
[55] Franks S，Robberts R，Hardy K，et al.Gonadotrophin regimens and oocyte quality inwomen with polycystic ovaries[J].Reprod Biomed Online，2003，6(2)：181–184
[56] Artini PG，Monti M，Matteucci C，et al.Vascular endothelial growth factor and basicfibroblast growth factor in polycystic ovary syndrome during controlled ovarianhyperstimulation[J].Gynecol Endocrinol，2006，22(8)：465-470
[57] Artini PG， Ruggiero M， Parisen Toldin MR，et al.Vascular endothelial growthfactor and its soluble receptor in patients with polycystic ovary syndromeundergoing IVF[J].Hum Fertil(Camb)，2009，12(1)：40-44
[58] Peitsidis P，Agrawal R.Role of vascular endothelial growth factor in women withPCO and PCOS：a systematic review[J].Reprod Biomed Online，2010，20(4)：444-452
[59]孙林，魏巍关，关咏梅.u-PA、U-PAR和PAI-1在睾酮联合hCG致多囊卵巢大鼠中的表达与意义[J].生殖与避孕，2006，26（10）：579-583
[60]申景欣，潘琼，薛敏，等.纤溶酶原激活因子及抑制因子在多囊卵巢综合征患者卵巢中的表达[J].第二军医大学学报，2010，12（2）：1309-1313
[61] Lindholm1A，Bixo M.Tissue plasminogen activator and plasminogen activatorinhibitor1in obese and lean patients with polycystic ovary syndrome[J]. GynecolEndocrinol，2010，26(10)：743-748
[62] Koiou E，Tziomalos K，Dinas K，et al.Plasma plasminogen activator inhibitor-1levelsin the different phenotypes of the polycystic ovary syndrome[J].Endocr J，2012，59(1)：21-9
[63] Lin YS，Tsai SJ，Lin MW，et al.Interleukin-6as an early chronic inflammatorymarker in Polycystic Ovary Syndrome with insulin receptor substrate-2polymorphism[J].Ame J Reprod mmunol，2011，66(6)：527-533
[64] Kim CH，Ahn JW，You RM，et al.Pioglitazone treatment decreases follicular fluidlevels of tumor necrosis factor-α and interleukin-6in patients with polycystic ovarysyndrome[J].Clin Exp Reprod Med.2011，38(2)：98–102
[65]游力，芦珂，赵跃然，等.血清抵抗素、C-反应蛋白及IL-6与PCOS的相关性研究[J].生殖与避孕，2010，30(5)：311-315
[66] Van Der Spuy ZM，Dyer SJ.The pathogenesis of infertility and early pregnancy lossin polycystic ovary syndrome[J].Best Pract Res Clin Obstet Gynaecol，2004，18(5)：755–771
[67]王秀霞，王天任，易晓芳，等.卵泡液中TNF-α和IFN-γ与卵丘颗粒细胞凋亡关系的探讨[J].生殖与避孕，2010，30(8)：519-522
[68]洪岭，滕晓明，李昆明，等.TNF-α在体外具有促进猪卵泡内膜细胞增殖的作用[J].华西医学，201025(1)：57-59
[69] Xiong YL，Liang XY，Yang X，et al.Low-grade chronic inflammation in theperipheral blood and ovaries of women with polycystic ovarian syndrome[J].Eur JObstet Gynecol Reprod Biol，2011，159(1)：148-150
[70]徐淑云，赵淑萍，马德花，等.PCOS病人血清TNF-α及IGF-1水平变化及意义[J].青岛大学医学院学报，2010，46(1)：45-48
[71] Pehlivanov B，Mitkov M.Serum leptin levels correlate with clinical and biochemicalindices of insulin resistance in women with polycystic ovary syndrome[J]，Eur JContracept Reprod Health Care，2009，14(2)：153-159
[72] Georgios A， Eleni K，Ioannis S，Vassilios L，et al.Serum and follicular fluid leptinlevels are correlated with human embryo quality[J].Reproduction，2005，130（1）：917–921
[73] Karamouti M，Kollia P，KallitsarisA，et al.Modulating effect of leptin on basal andfollicle stimulating hormone stimulated steroidogenesis in cultured human luteingranulosa cells[J].J Endocrinol，2009，32（5）：415-419
[74] Yildizhan R，Ilhan GA，Yildizhan B，et al.Serum retinol-binding protein4，leptin，and plasma asymmetric dimethylarginine levels in obese and nonobese youngwomen with polycystic ovary syndrome[J].Fertil Steril，2011，96(1)：246-251
[75] Seow KM，Juan CC，Hsu YP，Ho LT，Wang YY，Hwang JL.Serum and follicularresistin levels in women with polycystic ovarian syndrome during IVF-stimulatedcycles[J].Hum Reprod，2005，20(1)：117–121
[76]陆秀娥.多囊卵巢综合征患者颗粒细胞AQP9、RESISTIN表达及对体外受精结局的影响[D].浙江：浙江大学医学妇产科学生殖内分泌，2006
[77] Shen CJ，Tsai，EM，Lee JN，et al.The cocentrations of visfatin in the follicular fluidsof women undergoing controlled ovarian stimulation are correlated to the number ofoocytes retrieved[J].Fertil Steril，2010，93(6)：1844-1850
[78] Tan BK，Chen J，Digby JE，et al.Increased visfatin messenger ribonucleic acid andprotein levels in adipose tissue and adipocytes in women with polycystic ovarysyndrome：parallel increase in plasma visfatin[J].J Clin Endocrinol Metab，2006，91(12)：5022-5028
[79] Jongwutiwes T，Lertvikool S，Leelaphiwat S，et al.Serum visfatin in Asian womenwith polycystic ovary syndrome[J].Gynecol Endocrinol，2009，25(8)：536-542
[80] Chabrolle C，Tosca L，Ramé C，et al. Adiponectin increases insulin-like growthfactor-induced progesterone and estradiol secretion in human granulosacells[J].Fertil Steril，2009，92(6)：1988-1996
[81] Chappaz E，Albornoz MS，Campos D，et a1．Adiponectin enhances in vitrodevelopment of swine embryos[J].Domest Anjm Endocrin，2008，35(2)：198-207
[82] Louise MH，Leonhardt H，Kullberg J，et al.Adipose tissue has aberrant morphologyand function in PCOS：enlarged adipocytes and low serum adiponectin，but notcirculating sex steroids，are strongly associated with insulin resistance[J].J ClinEndocrinol Metab，2011，96(2)：304-311
[83]洪青青.多囊卵巢综合征患者子宫内膜和卵巢颗粒细胞脂联素及其受体表达的研究[D].上海，上海交通大学妇产科学，2009
[84] Matthias J，Kleinz，Anthony P，et al.Emerging roles of apelin in biology andmedicine[J].Pharmacology Therapeutics，2005，107(4)：198-211
[85]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[86] G ren K，Sa s z N，Noyan V，et al.Plasma apelin levels in patiets with polycysticovary syndrome [J]. J Turkish German Gynecol Assoc，2012，13(1)：27-31
[87] Chang CY，Tsai YC，Lee CH， et al.Lower serum apelin levels in women withpolycystic ovary syndrome [J].Fertil Steril，2011，95(8)：2520-2523
[88]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398
[89] Azziz R，Woods KS，Reyna R， et al. The prevalence and features of the polycysticovary syndrome in an unselected population [J].J Clin Endocrinol Metab，2004，89(6)：2745–2749
[90] Pasquali R.Obesity and androgens： facts and perspectives [J]. Fertil Steril，2006，85(5)：1319–1340
[91] Kuchenbecker WK， Groen H，Asselt SJ，et al.In women with polycystic ovarysyndrome and obesity， loss of intra-abdominal fat is associated with resumption ofovulation[J].Human Reprod.2011，26(9)：2505-2512
[1] Tatemoto K，Hosoya M，Habata Y，et al.Isolation and characterization of a novelendogenous peptide ligand for the human APJ receptor[J].Biochem Biophys ResCommun，1998，25(1)：471–476
[2] Takashi S，Naomichi K，Chiaki M，et a1.Apelin and APJ receptor expression ingranulosa and theca cells during different stages of follicular development in thebovine ovary： Involvement of apoptosis and hormonal regulation[J].Ani ReprodSci，2009，116(1-2)：28-37
[3]丁永刚，倪安民.脂肪因子apelin与代谢综合征[J].中国糖尿病杂志，2008，16(7)：444-447
[4] De Falco M，De Luca L，Onori N，et al.Apelin expression in normal humantissues[J].In Vivo，2002，16(5)：333-336
[5] Kawamata Y， Habata Y，Fukusumi S，et al.Molecular properties of apelin：tissuedistribution and receptor binding[J].Biochim Biophys Acta，2001，1538(2-3)：162-171
[6] Habata Y，Fujii R， Hosoya M.Apelin， the natural ligand of the orphan receptorAPJ，is abundantly secreted in the colostrum[J].Biochim Biophys Acta，1999，1452(1)：25-35
[7] De Falco M，Fedele V，Russo T，et al.Distribution of apelin，the endogenous ligandof the APJ receptor，in the lizard Podarcis sicula[J].J Mol Histol，2004，35(5)：521-527
[8] Katugampola SD，Maguire JJ，Matthewson SR，et al.[(125)I]-(Pyr(1))Apelin-13isa novel radioligand for localizing the APJ orphan receptor in human and rat tissueswith evidence for a vasoconstrictor role in man[J].Br J Pharmacol，2001，132(6)：1255–1260
[9] Reaux A， De Mota N， Skultetyova I，et al.Physiological role of a novelneuropeptide，apelin，and its receptor in the rat brain[J].J Neurochem，2001，77(4)：1085–1096
[10] Choe H， Farzan M， Konkel M， et al. The orphan seven-transmembrane receptorapj supports the entry of primary T-cell-line-tropic and dualtropic humanimmunodeficiency virus type1[J].J Virol，1998，72(7)：6113–6118
[11]巴宏军，陈红珊，杜敏联，马华梅，等.肥胖女童血清apelin、chemerin水平及其与胰岛素抵抗的关系[J].中国病理生理杂志，2011，27(10)：2014-2016
[12] Katarzyna Z，Oswiecimska J，Swietochowska E，et al.Assessment of serum apehnlevels in girls with anorexia nervosa[J]．J Clin Endocfinol Metab，2010，95(6)：2935-3941
[13] Isabelle CL， Michaela V， Daviaud D，et al.Effect of hypocaloric diet-inducedweight loss in obese women on plasma apelin and adipose tissue expression ofapelin and APJ[J].Eur J Endocrinol，2008158(6)：905-910
[14] Boucher J，Masri B， Daviaud D，et al.Apelin，a newly identified adipokineup-regulated by insulin and obesity[J].Endocrinology，2005，146(4)：1764-1771
[15] Kunduzova O， Alet1N， Delesque TN，et al.Apelin/APJ signaling system： apotential link between adipose tissue and endothelial angiogenic processes[J]. FAS--EB J，2008，(22）：4146-4154
[16] Li L，Yang G，Li Q．Changes and relations of circulating visfatin，apelin，and resistinlevels in normal，impaired glucose tolerance，and type2diabetic subjects[J]. ExpClin Endocrinol Diabet，2006，114(10)：544-548
[17]杨冬莲，黎英荣，连莲，等.肥胖2型糖尿病患者血清apelin的变化及其与血脂的关系研究[J].广西医科大学学报，2010，27(4)：565-568
[18] Dray C，Debard C，Jager J，et al.Apelin and APJ regulation in adipose tissue andskeletal muscle of type2diabetic mice and humans[J].Am J Physiol EndocrinolMetab，2010，298(6)：1161-1169
[19] Erdem G，Dogru T，Tasci I，et al.Apelin levels in newly diagnosed type2diabetesmellitus[J].Exp Clin Endocrinol Diabetes，2008，116(5)：289-292
[20] Zhang Y，Shen C，Li X，et al.Low plasma apelin in newly diagnosed type2diabetesin Chinese people[J].Diabetes Care，2009，32(12)：150-150
[21] Kadoglou N PE，Tsanikidis H，Kapelouzou A，et al.Effects of rosiglitazone andmetformin treatment on apelin，visfatin，and ghrelin levels in patients with type2diabetes mellitus[J].Metabolism，2010，59(3)：373-379
[22] Trayhurn P，Beattie JH．Physiological role of adipose tissue：white adipose tissue asan endocrine and secretory organ[J].Proc Nutr Soc，2001，60(3)：329-339
[23] Higuchi K，Masaki T，Gotoh K，et a1.Apelin，an APJ receptor ligand，regulates bodyadiposity and favors the mRNA expression of uncoupling proteinsi nmice[J].Endo--crinology，2007，148(6)：2690-2697
[24]李庆辉，杨天伦，谭碧峰.原发性高血压患者血清apelin-13与脂联素的相关性研究[J].医学临床研究，2011，28(2)：223-226
[25] Strazynska A，Bryl W，Hoffmann K，et al.The estimation of concentartion of apelin，insulin， fasting glucose and anthropometric factors in young hypertensivepatients[J].J Hypertens，2010，28(28)：187-187
[26] Monika PK，Tomasz K，Andrzej M，et al.Reduced circulating apelin in essentialhypertension and its association with cardiac dysfunction[J].J Hypertens，2011，29(5)：971-979
[27]廖予妹，乔福元．妊娠期高血压疾病患者胎盘组织中血管活性肽apelin的表达及其意义[J].中华妇产科杂志，2007，42(6)：382-386
[28] Ishida J，Hashimoto J，Hashimoto Y，et a1．Regulatory roles for APJ，a seventransmembrane receptor related to angiotensin type I receptor in blood pressure invivo[J]．Bio1Chem，2004，279(25)：26274-26279
[29] Hashinoto T，Kihara M，Ishida I，et a1．Apelin Stimulates myosin light chainphosphory1ation in vascular smooth muscle cells[J]．Arterioscler Thromb VascBiol，2006，26(6)：1267-1272
[30]李丽英，魏雪梅，王伟，等.Apelin/APJ系统研究新进展[J].重庆医学，2011，40（22）：2275-2279
[31]王芳，蒋玲，邓晓惠，等.多囊卵巢综合征患者血清apelin水平变化及相关因素分析[J].中国糖尿病杂志，2012，20(2)：95-99
[32] G ren K，Sa s z N，Noyan V，et al.Plasma Apelin levels in patiets with polycysticovary syndrome[J].J Turkish German Gynecol Assoc，2012，13(1)：27-31
[33] Chang CY， Tsai YC，Lee CH，et al.Lower serum apelin levels in women withpolycystic ovary syndrome[J].Fertil Steril，2011，95(8)：2520-2523
[34]罗爽，王继东，潘红娟，等.Apelin/APJ在PCOS大鼠卵巢中的表达及意义[J].生殖与避孕，2011，8(3)：392-398