益肾调周法干预卵巢早衰模型大鼠机制的实验研究
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摘要
目的：通过观察益肾调周法对卵巢早衰造模大鼠动情周期的影响；对卵巢甾体类固醇激素生成能力(E2、P、T)的影响；对卵巢储备能力的测评(FSH、LH、INHB、AMH);对体液免疫(AOAB)和神经免疫(β-EP)的调节；对卵巢颗粒细胞凋亡相关蛋白表达的干预(Bcl-2、Fas、Fas-I);对细胞免疫相关细胞因子(VEGF、TNF-α、 INF-γ)的调节;血清中血脂的变化(TC、TG)以及对卵巢组织形态学观察等项目,从神经-内分泌-免疫网络的整体性和协调性方面探讨该法干预卵巢早衰的机制,探讨其量效关系,为中医药防治POF的方法提供科学的依据。
方法：选SPF级健康雌性8周龄SD大鼠,通过阴道脱落细胞学涂片,选出有动情周期为受试动物,除空白组(A)外,其余大鼠均给予注射环磷酰胺致POF模型。环磷酰胺模型大鼠随机分为即模型对照组(B)、克龄蒙对照组(C)、滋肾填精汤高剂量组(D1)、滋肾填精汤低剂量组(D2)、温肾填精汤高剂量组(E1)、温肾填精汤低剂量组(E2)、益肾调周法高剂量组(F1)、益肾调周法低剂量组(F2)、益肾调周法高-低剂量组(G1)、益肾调周法低-高剂量组(G2),A,B组给予每日灌胃两次蒸馏水,中药各组实验当日冷开水配置所需动物灌胃浓度,低剂量,高剂量分别相当于临床等效药量的2、4倍,克龄蒙对照组采用相当于临床等效药量的4倍,每日灌胃一次。各组均连续用药20天。用药期间每日检测一次阴道脱落细胞,并记录。于实验末次给药后,实验大鼠断头采血,取血清。采用放射免疫计数仪测定血清中E2、T、P值;用酶联免疫测定试剂盒测定血清中FSH、LH、β-EP、 AOAB和VEGF、TNF-α、IFN-γ的表达;采用双抗体夹心酶联免疫吸附技术，检测AMH, INHB;氧化酶法测定血清TC、TG水平;取血后将大鼠,分离出下丘脑,酶联免疫测定试剂盒测定下丘脑β-EP水平;断头后迅速解剖同一侧卵巢,固定,脱水,石蜡包埋,连续切片,染色,最大剖面光镜下观察卵巢的各级卵泡数及黄体数的变化及病理改变;免疫组化法观察Bcl-2、Fas、Fas-L的蛋白表达。
结果：1.益肾调周法对动情周期的影响：益肾调周法高-低剂量(G1)能够有效改善POF模型大鼠的紊乱的动情周期,恢复正常周期。
2.益肾调周法对甾体类固醇激素的影响：益肾调周法高-低剂量组(G1)的血清E2,P值显著升高,而T值显著下降。
3.益肾调周法对卵巢储备能力的影响：益肾调周法高-低剂量组(G1)血清FSH, LH的水平显著降低,而血清AMH, INHB的表达增强。
4.益肾调周法对卵巢细胞免疫相关因子的影响：益肾调周法高-低剂量组((G1)血清TNF-α、FN-γ的表达显著降低,但是血清VEGF的表达增强不显著。
5.益肾调周法对卵巢颗粒细胞凋亡相关蛋白表达的影响：益肾调周法高-低剂量组(G1)卵巢颗粒细胞Bcl-2蛋白的阳性表达增强显著,Fas蛋白的表达增强,而使Fas-L蛋白表达减弱。
6.益肾调周法对体液免疫和神经免疫的影响：益肾调周法高-低剂量组(G1)血清AOAB浓度显著降低,血清和下丘脑的β-EP的含量显著提高。
7.益肾调周法对血脂的影响：益肾调周法高-低剂量组(G1)血清中TC和TG的水平显著降低。
8.益肾调周法对卵巢形态学的影响：益肾调周法高-低剂量组(G1)卵巢中初级卵泡,生长卵泡以及黄体的数量显著增加,炎性侵润减少。
结论：1.腹腔注射环磷酰胺,符合POF的诊断标准,成功造模。
2.益肾调周法高-低剂量(G1)有效改善紊乱的动情周期。
3.益肾调周法高-低剂量组(G1)能够调节卵巢颗粒细胞的甾体类固醇激素的生成并且能够提高卵巢的储备能力。
5.益肾调周法高-低剂量成功参与细胞免疫调节和体液免疫调节,加强对卵巢功能的神经调节,益肾调周法高-低剂量(G1)有效抑制卵巢细胞凋亡的发生。
8.益肾调周法高-低剂量组(G1)有效降低血脂并促进卵泡发育。
因此,益肾调周法能够通过调整NEI网络的多个环节以实现干预POF的综合效应。
Objective:Observation the effect of Yishen tune-periods method to oestrous cycle in POF model rats and ovarian steroid hormone generating capacity in POF model rats (E2、P、T); Observation the effect of Yishen tune-periods method to ovarian reserve capacity (FSH、LH、 INHB、AMH); Observation the moderate effect of Yishen tune-periods method to humoral immunity (AOAB) and neuroimmunology (P-EP); Observation the effect of Yishen tune-periods method to the apoptosis related proteins of granule cells (Bcl-2、Fas、Fas-1); Observation the moderate effect of Yishen tune-periods method to cellular immune related factors (VEGF、TNF-α INF-γ); Observation the change of Yishen tune-periods method to serum lipid (TC、TG) and Observation the effect of Yishen tune-periods method to ovarian morphology,in order to discuss interaction mechanism between POF and Yishen tune-periods method considering integrity and coordination of nerve-endocrine-immune network,and dose-effect relationship,it can provide the scientific basis in prevention and treatment POF with Chinese medicine.
Method:choose SPF female healthy8weeks SD rats, the vaginal exfoliated cytology of each rat was inspected to elect rats with normal estrous cycle for animal subjects, except the blank group (A), the rest of animal subjects are given by the injection of cyclophosphamide to establish POF model. POF model rats were randomly divided into namely model group (B),Climen group (C), Zishentianjing soup high-dose group (D1), Zishentianjing soup low-dose group (D2),Wenshentianjing soup high-dose group (E1),Wenshentianjing soup low-dose group(E2),Yishen tune-periods method high-dose group (F1),Yishen tune-periods method low-dose group (F2), Yishen tune-periods method high-low dose group (G1) and Yishen tune-periods method low-high dose group (G2). A, B group were poured distilled water into stomach twice daily, each Chinese traditional medicine group was poured into stomach with the cold water for required concentration, low dose, high doses were equivalent to the clinical equivalent quantity2,4times respectively, Climen group (C) is equivalent to the clinical equivalent4times of quantity, each group was poured once daily, Consecutive medication last20days. The vaginal exfoliated cytology of each rat was inspected and record.After the last medication, all rats were taken blood by decollation.after doing it,measure level of E2、P、T in serum using calculating instrument of radio-immunity method; detect level of FSH、LH、β-EP、AOAB and expression of VEGF、TNF-α IFN-yin serum using ELISA; detect level of AMH, INHB in serum using double antibody sandwich of ELISA; measure level of TC、TG in serum using CHO. Decollation to separate hypothalamus and measure content of β-EP with hypothalamus using ELISA;spaying and fixation with paraffins,slicing,red dying to observe quantity of follicle at all levels and pathological change of follicle;measure protein expression of Bcl-2、Fas、Fas-L in serum using immunohistochemical method
Result:1. The effect of Yishen tune-periods method to oestrous cycle:Disorder oestrous cycle was improved in Yishen tune-periods method high-lowdose group (G1) dramatically.
2. The effect of Yishen tunc-periods method to ovarian steroid hormone generating capacity:The level of E2, P in serum of Yishen tune-periods method high-lowdose group (G1) was elevated dramatically and the level of T in same group was reduced dramatically.
3. The effect of Yishen tune-periods method to ovarian reserve capacity:The level of FSH、LH in serum of Yishen tune-periods method high-low dose group (G1) drops dramatically,the expression of INHB、 AMH in serum was strengthened apparently.
4. The moderate effect of Yishen tune-periods method to cellular immune related factors:The expression of TNF-αand INF-ydrops apparently in serum of Yishen tune-periods method high-low dose group (G1), and the expression of VEGF are strengthened in same group indistinctively.
5.The effect of Yishen tune-periods method to the apoptosis related proteins of granule cells:Not only the expression of Bcl-2in serum of Yishen tune-periods method high-low dose group (G1) was strengthened distinctively, but also Fas did, in the contrary, the expression of Fas-1drops apparently.
6.The moderate effect of Yishen tune-periods method to humoral immunityand neuroimmunology:The concentration of AOAB in serum of Yishen tune-periods method high-low dose group (G1) drop apparently,and the concentration of (3-EP in serum and hypothalamus are elevated apparently in same group.
7. The change of Yishen tune-periods method to serum lipid:The level of both TC and TG drop apparently in serum of Yishen tune-periods method high-low dose group (G1).
8.The effect of Yishen tune-periods method to ovarian morphology:The quantity of primary follicle,growing follicle and corpus luteum are incr-eased apparently in ovary of Yishen tune-periods method high-low dose group (G1),and inflammatory assault embellish are redused apparently in same group.
Conclusion:1. It is succeeding by the injection of cyclophosphamide to establish POF model.
2. Yishen tune-periods method with high-low dose can improve the disorder oestrous cycle.
3. Yishen tune-periods method with high-low dose can adjust ovarian steroid hormone generating capacity.
4. Yishen tune-periods method with high-low dose can improve ovarian reserve capacity.
5. Yishen tune-periods method with high-low dose can participate in immune adjustment and follicle growth.
6. Yishen tune-periods method with high-low dose can control the apoptosis of ovarian granulose cell.
7. Yishen tune-periods method with high-low dose can weaken immune response and enhance neuroregulation act on ovary.
8. Yishen tune-periods method with high-low dose can reduce the contect of serum lipid.
9. Yishen tune-periods method with high-low dose can encourage follicular development.
Therefore, Yishen tune-periods method can adjust the NEI network through multiple links in order to realize the comprehensive effect on the intervention of POF.
方法：选SPF级健康雌性8周龄SD大鼠,通过阴道脱落细胞学涂片,选出有动情周期为受试动物,除空白组(A)外,其余大鼠均给予注射环磷酰胺致POF模型。环磷酰胺模型大鼠随机分为即模型对照组(B)、克龄蒙对照组(C)、滋肾填精汤高剂量组(D1)、滋肾填精汤低剂量组(D2)、温肾填精汤高剂量组(E1)、温肾填精汤低剂量组(E2)、益肾调周法高剂量组(F1)、益肾调周法低剂量组(F2)、益肾调周法高-低剂量组(G1)、益肾调周法低-高剂量组(G2),A,B组给予每日灌胃两次蒸馏水,中药各组实验当日冷开水配置所需动物灌胃浓度,低剂量,高剂量分别相当于临床等效药量的2、4倍,克龄蒙对照组采用相当于临床等效药量的4倍,每日灌胃一次。各组均连续用药20天。用药期间每日检测一次阴道脱落细胞,并记录。于实验末次给药后,实验大鼠断头采血,取血清。采用放射免疫计数仪测定血清中E2、T、P值;用酶联免疫测定试剂盒测定血清中FSH、LH、β-EP、 AOAB和VEGF、TNF-α、IFN-γ的表达;采用双抗体夹心酶联免疫吸附技术，检测AMH, INHB;氧化酶法测定血清TC、TG水平;取血后将大鼠,分离出下丘脑,酶联免疫测定试剂盒测定下丘脑β-EP水平;断头后迅速解剖同一侧卵巢,固定,脱水,石蜡包埋,连续切片,染色,最大剖面光镜下观察卵巢的各级卵泡数及黄体数的变化及病理改变;免疫组化法观察Bcl-2、Fas、Fas-L的蛋白表达。
结果：1.益肾调周法对动情周期的影响：益肾调周法高-低剂量(G1)能够有效改善POF模型大鼠的紊乱的动情周期,恢复正常周期。
2.益肾调周法对甾体类固醇激素的影响：益肾调周法高-低剂量组(G1)的血清E2,P值显著升高,而T值显著下降。
3.益肾调周法对卵巢储备能力的影响：益肾调周法高-低剂量组(G1)血清FSH, LH的水平显著降低,而血清AMH, INHB的表达增强。
4.益肾调周法对卵巢细胞免疫相关因子的影响：益肾调周法高-低剂量组((G1)血清TNF-α、FN-γ的表达显著降低,但是血清VEGF的表达增强不显著。
5.益肾调周法对卵巢颗粒细胞凋亡相关蛋白表达的影响：益肾调周法高-低剂量组(G1)卵巢颗粒细胞Bcl-2蛋白的阳性表达增强显著,Fas蛋白的表达增强,而使Fas-L蛋白表达减弱。
6.益肾调周法对体液免疫和神经免疫的影响：益肾调周法高-低剂量组(G1)血清AOAB浓度显著降低,血清和下丘脑的β-EP的含量显著提高。
7.益肾调周法对血脂的影响：益肾调周法高-低剂量组(G1)血清中TC和TG的水平显著降低。
8.益肾调周法对卵巢形态学的影响：益肾调周法高-低剂量组(G1)卵巢中初级卵泡,生长卵泡以及黄体的数量显著增加,炎性侵润减少。
结论：1.腹腔注射环磷酰胺,符合POF的诊断标准,成功造模。
2.益肾调周法高-低剂量(G1)有效改善紊乱的动情周期。
3.益肾调周法高-低剂量组(G1)能够调节卵巢颗粒细胞的甾体类固醇激素的生成并且能够提高卵巢的储备能力。
5.益肾调周法高-低剂量成功参与细胞免疫调节和体液免疫调节,加强对卵巢功能的神经调节,益肾调周法高-低剂量(G1)有效抑制卵巢细胞凋亡的发生。
8.益肾调周法高-低剂量组(G1)有效降低血脂并促进卵泡发育。
因此,益肾调周法能够通过调整NEI网络的多个环节以实现干预POF的综合效应。
Objective:Observation the effect of Yishen tune-periods method to oestrous cycle in POF model rats and ovarian steroid hormone generating capacity in POF model rats (E2、P、T); Observation the effect of Yishen tune-periods method to ovarian reserve capacity (FSH、LH、 INHB、AMH); Observation the moderate effect of Yishen tune-periods method to humoral immunity (AOAB) and neuroimmunology (P-EP); Observation the effect of Yishen tune-periods method to the apoptosis related proteins of granule cells (Bcl-2、Fas、Fas-1); Observation the moderate effect of Yishen tune-periods method to cellular immune related factors (VEGF、TNF-α INF-γ); Observation the change of Yishen tune-periods method to serum lipid (TC、TG) and Observation the effect of Yishen tune-periods method to ovarian morphology,in order to discuss interaction mechanism between POF and Yishen tune-periods method considering integrity and coordination of nerve-endocrine-immune network,and dose-effect relationship,it can provide the scientific basis in prevention and treatment POF with Chinese medicine.
Method:choose SPF female healthy8weeks SD rats, the vaginal exfoliated cytology of each rat was inspected to elect rats with normal estrous cycle for animal subjects, except the blank group (A), the rest of animal subjects are given by the injection of cyclophosphamide to establish POF model. POF model rats were randomly divided into namely model group (B),Climen group (C), Zishentianjing soup high-dose group (D1), Zishentianjing soup low-dose group (D2),Wenshentianjing soup high-dose group (E1),Wenshentianjing soup low-dose group(E2),Yishen tune-periods method high-dose group (F1),Yishen tune-periods method low-dose group (F2), Yishen tune-periods method high-low dose group (G1) and Yishen tune-periods method low-high dose group (G2). A, B group were poured distilled water into stomach twice daily, each Chinese traditional medicine group was poured into stomach with the cold water for required concentration, low dose, high doses were equivalent to the clinical equivalent quantity2,4times respectively, Climen group (C) is equivalent to the clinical equivalent4times of quantity, each group was poured once daily, Consecutive medication last20days. The vaginal exfoliated cytology of each rat was inspected and record.After the last medication, all rats were taken blood by decollation.after doing it,measure level of E2、P、T in serum using calculating instrument of radio-immunity method; detect level of FSH、LH、β-EP、AOAB and expression of VEGF、TNF-α IFN-yin serum using ELISA; detect level of AMH, INHB in serum using double antibody sandwich of ELISA; measure level of TC、TG in serum using CHO. Decollation to separate hypothalamus and measure content of β-EP with hypothalamus using ELISA;spaying and fixation with paraffins,slicing,red dying to observe quantity of follicle at all levels and pathological change of follicle;measure protein expression of Bcl-2、Fas、Fas-L in serum using immunohistochemical method
Result:1. The effect of Yishen tune-periods method to oestrous cycle:Disorder oestrous cycle was improved in Yishen tune-periods method high-lowdose group (G1) dramatically.
2. The effect of Yishen tunc-periods method to ovarian steroid hormone generating capacity:The level of E2, P in serum of Yishen tune-periods method high-lowdose group (G1) was elevated dramatically and the level of T in same group was reduced dramatically.
3. The effect of Yishen tune-periods method to ovarian reserve capacity:The level of FSH、LH in serum of Yishen tune-periods method high-low dose group (G1) drops dramatically,the expression of INHB、 AMH in serum was strengthened apparently.
4. The moderate effect of Yishen tune-periods method to cellular immune related factors:The expression of TNF-αand INF-ydrops apparently in serum of Yishen tune-periods method high-low dose group (G1), and the expression of VEGF are strengthened in same group indistinctively.
5.The effect of Yishen tune-periods method to the apoptosis related proteins of granule cells:Not only the expression of Bcl-2in serum of Yishen tune-periods method high-low dose group (G1) was strengthened distinctively, but also Fas did, in the contrary, the expression of Fas-1drops apparently.
6.The moderate effect of Yishen tune-periods method to humoral immunityand neuroimmunology:The concentration of AOAB in serum of Yishen tune-periods method high-low dose group (G1) drop apparently,and the concentration of (3-EP in serum and hypothalamus are elevated apparently in same group.
7. The change of Yishen tune-periods method to serum lipid:The level of both TC and TG drop apparently in serum of Yishen tune-periods method high-low dose group (G1).
8.The effect of Yishen tune-periods method to ovarian morphology:The quantity of primary follicle,growing follicle and corpus luteum are incr-eased apparently in ovary of Yishen tune-periods method high-low dose group (G1),and inflammatory assault embellish are redused apparently in same group.
Conclusion:1. It is succeeding by the injection of cyclophosphamide to establish POF model.
2. Yishen tune-periods method with high-low dose can improve the disorder oestrous cycle.
3. Yishen tune-periods method with high-low dose can adjust ovarian steroid hormone generating capacity.
4. Yishen tune-periods method with high-low dose can improve ovarian reserve capacity.
5. Yishen tune-periods method with high-low dose can participate in immune adjustment and follicle growth.
6. Yishen tune-periods method with high-low dose can control the apoptosis of ovarian granulose cell.
7. Yishen tune-periods method with high-low dose can weaken immune response and enhance neuroregulation act on ovary.
8. Yishen tune-periods method with high-low dose can reduce the contect of serum lipid.
9. Yishen tune-periods method with high-low dose can encourage follicular development.
Therefore, Yishen tune-periods method can adjust the NEI network through multiple links in order to realize the comprehensive effect on the intervention of POF.
引文
[1]Luborsky JL,Meyer P,Sowers MF,et al.Premature menopause in a multi-ethnic population study of the menopause transition Hum Reprod[J].2003, Jan; 18(1):199-206.
[2]Laml T, Schulz-Lobmeyr I, Obruca A,et al. Premature ovarian failure:etiology and prospects[J].Gynecol Endocrinol,2000,Aug; 14(4):292-302.
[3]Kalantaridou SN, Davis SR, Nelson LM. Premature ovarian failure[J].Endocrinol Metab Clin North Am,1998,Dec;27(4):989-1006.
[4]Santoro N. Mechanisms of premature ovarian failure. [J].An Endocrinol 2003,64:87-92.
[5]Timmreck LS, Reindollar RH:Contemporary issues in primary amenorrhea. [J].Obstetgynecol Clin North Am 2003,30:287-302.
[6]Coulam CB, Adamson SC, Annegers JF.Incidence of premature ovarian failure [J].Obstet Gynecol,1986,Apr;67(4):604-6.
[7]司徒仪主编.中西医结合妇产科学[M].北京：北京科学技术出版社,2003：155.
[8]SHYR-Yeu Lin, John R Morrison, David J, et al. Regulation of ovarian function by the TGF-superfamily and follistatin [J].Reproduction,2003,126:133-148.
[9]Welt. Female reproductive aging ismarked by decreased secretion of dimeric inhibin[J]. Clin EndocrinolMetab,1999,84(1):105
[10]La Marca A, Stabile G, Tirelli A, et al. Anti-mullerian hormonemeasurement on any day of the menstrual cycle strongly predicts o-varian response in assisted reproductive technology [J]. Hum Re-prod,2007,22(3):766-771.
[11]Andersen CY,ByskovAG.Estradiol and regulation ofantiMullerian hormone, inhibin-A, and inhibin-B secretion:analysis of small antral and preovulatory human folliclesfluid[J]. J Clin EndocrinolMetab,2006,91:4064-4069
[12]KevenaarME,Themmen AP,Laven JS, et a.l Anti-Mu-llerian hormone and anti-Mullerian hormone type 11 receptor polymorphisms are associatedwith follicularphase estradiol levels in normo-ovulatory women [J]. Hum Reprod,2007,22:1547-1554
[13]张敏慧 丛晶者等.抗苗勒管激素与卵巢功能[J]国际生殖健康／计划生育杂志,2009,28(5)：281-286.
[14]Bussolino F,Mantovani A,Persico G. Molecular mechanisms of blood vessel formation[J]. TIBS,1997,221251
[15]GleieherN,WeglloferA, Barad DH.A Pilot study of Premature ovarian seneseenee: Ⅱ.Different genotype and Pllenotype for genetic and autoimmune etiologies.FertilSteri, Ⅰ 2008,3(7):203-205.
[16]DongL,JiangL,MenW, et al.Preventive and therapeutic effects of BushenHuoxue Recipe on autoimmune premature ovarian failure in mice.ZhongxiyiJieHexueBao,2008,6(3):294-297.
[17]Gregoraszczuk EL,Slomczynska M. Beta-endorphin inhibition ofprogesterone secretion by porcine granulosa cells during follicledevelopment[J].Reprod NutrDev,1998,38(3):227-234.
[18]Besedovsky H, Sorkin E. Network of immune-neuroendocrine interactions[J].Clin Exp Immunol,1977,Jan; 27(1):1-12.
[19]陈晓萍,徐远扬.神经、内分泌、免逸网络的通用生物学语言[J].自然杂志,2002,24(4)：194.
[20]郭蕾,王永炎,张志斌.关于证候概念的诊释[J1.北京中医药大学学报,2003,26(2)：5-8.
[21]张焱,回世洋,张庆荣.Z2医五脏调控系统与神经内分泌免逸调节网络的关系[J].中华中医药学刊,2009,27(1)：188-189.
[22]金婧,阮祥燕.卵巢早衰的病因研究进展.[J],实用妇产科杂志,2007,23(3)：142-145.
[23]Kauffman RP, Castracane VD.Premature ovarian failure asociated with autoimmune polyglandular syndrome:pathophysiological mechanisms and future fertility. [J].Women's Health(Larchmt),2003,12(5):513-520.
[24]杨业洲.卵巢早衰.[M].北京：人民卫生出版社,2004,102-108.
[25]孙晓溪,李大金,王明雁,等.自身免疫性卵巢功能损伤与TH1/TH2型细胞因子表达关系的研究.[J].中国免疫学杂志,2001,17(2)：96-98.
[26]Chernyshov VP, RadyshTV, Gura Ⅳ, et al.Immune disorders in women with premature ovarian failure in initial period. [J].Am J Reprod Immunol.2001,46(3):220-225.
[27]王一峰,欧汝强,杨宁,等.卵巢早衰患者血消抗透明带抗体和肿瘤坏死因子-α,γ-干扰素及白细胞介素-2的分析.[J].生殖医学杂志,2002,11(1)：7-10.
[28]Monnier Barbarino P, Forges T, Faureg C.gonadal antibodies interfeting with female reproduction. [J].Best-Pract-Res-Clin-Endocrinol-Metab.2005,19(1):135-148.
[29]DoldiN, Belvisi L, Bassan M, et al.Premature ovarian failure:steroid synthesis and autoimmunity. [J].gynecol Endocrinol.1998,12(1):23-28.
[30]赵建英,高晓秋,史樱.35例卵巢早衰患者雌二醇与睾酮水平分析.[J].生殖医学杂志,2000,9(6)：355-356.
[31]Bakalov VK, Vanderhoof VH, Bondy CA, et al. Adrenal antibodies detect asymptomatic auto-immune adrenal insufficiency in young women with spontaneous premature ovarian failure. [J].Hum Reprod,2002,17(8):2096-2100.
[32]李珂,补肾健脾调肝法治疗卵巢早衰及对POF模型大鼠血清INHB、VEGF的影响.广州中医药大学2006届博士学位论文,2006,7.
[33]杨桂艳,王一理,曹绩孙等.卵巢早衰患者外周血CD4+、THI/TH2细胞分布研究.生殖医学杂志,1998；7(4)：227—230.
[34]Arif S, Underhill JA, Donaldson P, et al. Human leukocyte antigen-DQB1*genotypes aspartate at position 57 are associated with 3β-Hydroxysteroid dehydrogenase auto-immunity in premature ovarian failure. [J].Clin Endocrinol Metab,1999,84(3): 1056-1060.
[35]Reimand K, Peterson P, Hyoty H.3beta-hydroxysteroid dehydrogenase autoantibodies are rare in premature ovarian failure. [J].Clin Endocrinol Metab.2000 Jun; 85(6):2324-6.
[36]Zhi-Bin Tong, Lawrence M. Nelson. A Mousegene Encoding an Oocyte Antigen Associated with Autoimmune Premature Ovarian Failure. [J]. Endocrinology.1999, 140(8):3720-3726.
[37]Rhim SH, Millar SE, RobeyF, et al. Autoimmune disease of the ovary induced by a ZP3 peptide from the mouse zona pellucid. [J]. Clin Invest.1992 Jan,89(1):28-35.
[38]Naz RK, Thurston D, Santoro N. Circulating tumor necrosis factor (TNF)-alpha in normally cycling women and patients with premature ovarian failure and polycystic ovaries. [J]. Am J Reprod Immunol.1995 Sep; 34(3):170-5.
[39]Giovannigrasso, Michela Muscettola. Possible Role of Interferon-Gamma in Ovarian Function [M]. New York:Annals of the New York Academy of Sciences,1992:650(15): 191-196.
[40]Russell P. The clinicopathological features of primary ovarian failure. [J]. Verh Dtschges Pathol.1997,81:197-209.
[41]谢端筱,欧汝强,杨宁等.卵巢早衰与细胞因子TNF-a, IFN-y和IL-2的相关性研究.[J].广东医学,1999,20(11)：831-832.
[42]Felice Petraglia, Beda Hartmann, Stefano Luisi, et al. Low levels of serum inhibin A and inhibin B in women with hypergonadotropic amenorrhea and evidence of high levels of activin A in women with hypothalamic amenorrhea. [J]. Fertility and Sterility,1998, 70(5):907-912.
[43]陈蔚,雷小敏.卵巢早衰病因学研究进展.[J].生殖与避孕,2004,24(1)：43-7.
[44]李洁,杨菁.卵巢早衰动物模型建立的研究进展.[J].生殖与避孕,2006,26(9)：550-53.
[45]Yang PM, Chen HC, Tsai JS, et al. Cadmium induces Ca2+-dependent necrotic cell death through calpain-triggered mitochondrial depolarization and reactive oxygen species-mediated inhibition of nuclear factor-kappaB activity. [J]. Chem Res Toxicol. 2007,20(3):406-15.
[46]Sharabidze N, Burkadzeg, Sabakhtarashvili M. Cell adhesion and apoptosis in ovarian stromal hyperplasia and hyperthecosis. [N].georgian Med News.2006, (131):33-7.
[47]Depalo R, Nappi L, Loverrog, et al. Evidence of apoptosis in human primordial and primary follicles. [J]. Hum Reprod.2003,18(12):2678-82.
[48]王雪峰,何援利.凋亡调节基因bcl-2/bax与卵巢早衰的关系.[J],生殖与避孕,2008,28(8)：487-90.
[49]桑海滨.卵巢早衰的中西医诊治.贵阳中医学院学报,2006,28(4)：41-43.
[50]Morita Y, Perezgl, Paris F, et al. Oocyte apoptosis is suppressed by disruption of the acid sphingomyelinasegene or by sphingosine-1-phosphate therapy. [J].Nat Med.2000, 6(10):1109-14.
[51]Chauhan P, Sodhi A, Tarang S. Cisplatin-treated murine peritoneal macrophages induce apoptosis in L929 cells:role of Fas-Fas ligand and tumor necrosis factor-tumor necrosis factor receptor 1. [J]. Anticancer Drugs.2007,18(2):187-96.
[52]艾红,胡海燕,曹攒孙.自然排卵周期中子宫卵巢血流与性激素、血管内皮生长因子的关系.[J].中国医学影像杂志,2004,12(4)：286-88.
[53]Agrawal R, ConwaygS, Sladkevicius, et al. Serum vascular endothelialgrowth factor (VEGF) in the normal menstrual cycle:association with changes in ovarian and uterine Doppler blood flow. [J]. Clin Endocrinol (Oxf).1999,50(1):101-06.
[54]Seang Lin Tan, Jamal Zaidi, Stuart Campbell, et al. Blood flow changes in the ovarian and uterine arteries during the normal menstrual cycle. [J]. American Journal of Obstetrics &gynecology.1996,175(3):625-31.
[55]van Kasteren YM, Hundscheid RD, Smits AP, et al. Familial idiopathic premature ovarian failure:an overrated and underestimatedgenetic disease?. [J]. Hum Reprod.1999, 14(10):2455-59.
[56]Vegetti W, Marozzi A, Manfredini E, et al. Premature ovarian failure. [J]. Mol Cell Endocrinol.2000,161(1-2):53-7.
[57]Progetto Menopausa Italia Studygroup. Premature ovarian failure:frequency and risk factors among women attending a network of menopause clinics in Italy. [R]. BJOG.2003, 110(1):59-63.
[58]Testag, Chiaffarino F, Vegetti W, et al. Case-control study on risk factors for premature ovarian failure. [J].gynecol Obstet Invest.2001,51(1):40-3.
[59]Frisch RE. Body fat, menarche, fitness and fertility. [J]. Hum Reprod.1987,2(6): 521-33.
[60]Harlow BL, Signorello LB. Factors associated with early menopause. [J]. Maturitas, 2003,5(1):3.
[61]徐苓,宋亦军.卵巢早衰的临床表现和诊断标准.[J].实用妇产科杂志,2003,19(4)：195-196.
[62]张凇文,王军华,周红等.北京地区女性绝经期相关年龄相关因素的调查分析[C].第一次绝经相关问题研讨会论文汇编,2001,11(4)：164.
[63]许小凤,范春,包广勤等.卵巢早衰与卵巢储备功能下降的相关发病因素探讨[J].2011,11(4)：400-403.
[64]Luborsky JL, Meyer P, Sowers MF, et al. Premature menopause in a multi-ethnic population study of the menopause transition. [J]. Hum Reprod,2003,18(1):199-206.
[65]Kalantaridou SN, Nelson LM. Premature ovarian failure is not premature menopause. [M]. Ann N Y Acad Sci.2000,900:393-402.
[66]Ataya KM, McKanna JA, Weintraub AM, et al. A luteinizing hormone-releasing hormone agonist for the prevention of chemotherapy-induced ovarian follicular loss in rats. [J]. Cancer Res.1985 Aug; 45(8):3651-6.
[67]付霞霏,何援利.化疗所致卵巢早衰动物模型的建立.[J]广东医学,2008,29(12)：1952-1954.
[68]Yu X, Kamijima M, Ichiharag, et al.2-Bromopropane causes ovarian dysfunction by damaging primordial follicles and their oocytes in female rats. [J]. Toxicol Appl Pharmacol. 1999 Sep 15; 159(3):185-93.
[69]艾浩,牛建昭,薛晓等.顺铂致小鼠卵巢功能早衰肝肾阴虚证机制研究,[J]北京中医药大学学报,2006,29(6)：401-403.
[70]高慧,杨涓,韩冰,等.药物型卵巢早衰动物模型的研究.[J]国医论坛,2007,22(3)：22-26.
[71]付莉,赵怡璇,李守柔.卵巢早衰实验动物模型的建立[J].生殖医学杂志,2006,15(3)：179-183.
[72]林建华,严隽鸿,林其德.自身免疫性卵巢衰退小鼠动物模型的建立[J].上海医学,1999,12(12)：747-748.
[73]Bagavant H, Adams S, Terranova P, et al. Autoimmune ovarian inflammation triggered by pro inflammatory (Th1) T cells is compatible with normal ovarian function in mice. [J]. Biol Reprod.1999 Sep; 61(3):635-42.
[74]司卉良,陈友香,侯安继,等.补肾汤对激素致更年期肾阳虚证大鼠卵巢功能的影响.[J].中药药理与临床,2005,21(2)：44-45.
[75]Laissue P, Christin-Maitre S, Touraine P, et al. Mutations and sequence variants ingDF9 and BMP 15 in patients with premature ovarian failure. [J]. Eur J Endocrinol.2006 May; 154(5):739-44.
[76]Kelkar RL, Meherji PK, Kadam SS, et al. Circulating auto-antibodies against the zona pellucida and thyroid microsomal antigen in women with premature ovarian failure. [J]. J Reprod Immunol.2005 Jun; 66(1):53-67.
[77]Omoto Y, Lathe R, Warner M, et al. Early onset of puberty and early ovarian failure in CYP7B1 knockout mice. [J]. Proc Nat1 Acad Sci U S A.2005 Feb 22; 102(8):2814-9. Epub 2005 Feb 14.
[78]Takai Y, Canning J, PerezgI, et al. Bax, caspase-2, and caspase-3 are required for ovarian follicle loss caused by 4-vinylcyclohexene diepoxide exposure of female mice in vivo. [J]. Endocrinology.2003 Jan; 144(1):69-74.
[79]Yu WJ, Lee BJ, Nam SY, et al. Modulating effects of Koreanginseng saponins on ovarian function in immature rats. [J]. Biol Pharm Bull.2003 Nov; 26(11):1574-80.
[80]Olff M. Stress, depression and immunity:the role of defense and coping styles. [J]. Psychiatry Res.1999 Jan 18,85(1):7-15.
[81]连晓媛,丁岩,陈奇,等.重复制动应激对雌性大鼠卵巢功能的影响.[J].中药新药与临床药理,2004,15(6)：373-376.
[82]Bandyopadhyay S, Chakrabarti J, Banerjee S, et al. Prenatal exposure to highgalactose adversely affects initialgonadal pool ofgerm cells in rats. [J]. Hum Reprod.2003 Feb, 18(2):276-82.
[83]李洁,杨菁,陈嫒,等.利用不同浓度D(+)半乳糖建立卵巢早衰动物模型.[J].武汉大学学报(医学版),2006,27(5)：614-619.
[84]莫中福,殷秀玲,尹青,等DahlS高血压大鼠卵巢细胞凋亡相关因素的实验研究.[J].生殖与避孕,2002,22(5)：263-265.
[85]张晓金,归绥琪,徐丛剑.补肾复方调控更年期大鼠卵巢功能的实验研究.[J].现代中西医结合杂志,2003,12(19)：2044-2047.
[86]张绍芬,谢倩,哈灵侠,等.戊酸雌二醇对更年期大鼠脾细胞凋亡及卵巢功能的作用.[J].生殖医学杂志,2006,15(6)：388-392.
[87]蔡少锋,康幼雯.中西医结合人工周期疗法抗卵巢早衰的临床探讨.[J].中国民间疗法,1 997(6)：20.
[88]王锡珍,赖毛华.中医对卵巢早衰的认识和证治体会.[J]浙江中医学院学报,2003,27(5)：21-22
[89]王菊素.田淑霄教授对卵巢早衰与肾虚血瘀相关性的见解及治疗经验.[J].陕西中医,2003,11(24)：1015.
[90]朱玲,.罗颂平.卵巢早衰的病因病机及其论治.[J].中医药学刊,2003,21(1)：143-144.
[91]夏天.卵巢早衰的中西医结合研究进展.[J].辽宁中医杂志,2004,31(8)：707-708.
[92]黄健萍.卵巢早衰的中医治疗述要.[J].辽宁中医学院学报,2004,6(2)：126.
[93]张玉珍,史云.试论中医药防治卵巢早衰的思路与方法.[J]中医杂志,2005,46,(增刊)：116-117.
[94]魏智慧,邓琳文.浅议从肾论治卵巢早衰.[J].甘肃中医,2007,20(9)：39-40.
[95]刘霞,谢萍.浅析卵巢早衰的病机和证治.[J].江西中医学院学报,2007,19(4)：18-19.
[96]高慧,杨涓,李巧芬.中医学对卵巢早衰病因病机的认识.[J].甘肃中医,2007,20(8)：47-48.
[97]蔡立荣,李大金,孙晓溪,等.补肾活血方对小鼠实验性卵巢早衰防治作用的研究.[J].中国中西医结合杂志,2001,21(2)：126.
[98]钟雪梅,周灵,杨家林.补经合剂促卵泡发育和排卵的实验研究.江苏中医,1998,19(3)：45-46.
[99]董莉,谈嫒,平瑜佳等.不同治法方药治疗小鼠自身免疫性卵巢功能衰退的比较研究.[J]中西医结合学报2010,8(1)：86-89.
[100]张军武.归肾丸对大鼠卵巢早衰防治作用的实验研究.[J]中国现代医药杂志2008,10(6)：79-80.
[101]孙永忠,程泾,石其贤.滋肾益冲抗衰汤治疗卵巢早衰实验研究.[J]实用中医药杂志,2008,24(2)：71-73.
[102]朱玲,罗颂平,许丽绵等.左归丸对免疫性卵巢早衰小鼠抗卵巢抗体的影响.现代中西医结合杂志,[J]2006,15(4)：435-438.
[103]朱玲,罗颂平,许丽绵等.左归丸对免疫性卵巢早衰小鼠卵巢Fas、Fas-L表达的影响,[J]江西中医学院学报2008,20(1)：52-55.
[104]杨兰英.六味地黄丸治疗卵巢早衰的实验研究.[J].山西中医学院学报,2002,3(1)：21
[105]梁欣韫,孙晓芳,牛建昭等.育泡饮对自身免疫性卵巢损伤小鼠卵巢颗粒细胞的超微结构影响.[J]中华中医药学刊,2008,26(3)：502-504.
[106]李军,王翠霞,陈莹等.补肾复方延缓雌性大鼠卵巢早衰的实验研究.[J]辽宁中医杂志2007,3(5)：681-682.
[107]高慧,杨涓,夏天等.补肾调冲方对卵巢早衰大鼠卵巢相关细胞因子TNF-α、IFN-γ蛋白和基因表达的影响.[J]国医论坛2008,23(2)：32-39.
[108]李沛霖,侯莉莉,杜永红.补肾调冲含药血清对大鼠卵巢细胞体外分泌功能的影响.[J].时珍国医国药,2006,17(3)：354-355.
[109]滕秀香.122例卵巢早衰患者中医证候分析及致病因素调查[J] Chinese Journal of Information on TCM,2008,15(4):18-20.
[110]张朝辉,张亚辉.阳和汤治疗卵巢早衰症治验.[J].北京中医,2001,17(4)：16
[111]陈秀芳.辨证治疗卵巢早衰22例.[J].四川中医,2000,18(1)：35
[112]梁雪雯,苏建芬,王迪华,等.逍遥助卵煎治疗卵巢早衰30例.[J].上海中医药杂志,2006,40(2)：31-32.
[113]许昕,藤秀香,佟庆.补肾养血调经法治疗有卵泡型卵巢早衰.[J].北京中医,2001,(4)：21
[114]魏丹,肖秋霞.补肾养血调经法治疗卵巢早衰12例.[J].江西中医药,2008,39(30)：43-44.
[115]种丽群.补肾活血化淤法治疗卵巢早衰24例.[J].实用中医药杂志,2006,22(11)：675。
[116]兰英,魏巍.柴瑞霭治疗卵巢早衰验案举隅.[J].山西中医,2001,17(4)：5
[117]钱菁.夏桂成妇科病症验案举隅.[J].辽宁中医杂志,2005,32(11)：1198-1199.
[118]李云君,姜惠中.姜惠中治疗卵巢早衰的经验.[J].湖北中医杂志,2006,28(6)：28-29.
[119]周华,赵春梅.中西医治疗卵巢早衰临床研究述略.[J].中医药学刊,2004,22<5)：903.
[120]秦薇,周梅玲,陈祥云.中药人工周期疗法治疗卵巢功能早衰45例.[J].国医论坛,2004,19(1)：30-33.
[121]池雷,夏桂成.夏桂成教授调周疗法治疗月经病述要.实用医技杂志,2008,15(33)：4914.
[122]武淑慧,庆璐,崔晓萍.崔晓萍教授循期阴阳序贯疗法治疗卵巢早衰.[J]陕西中医学院学报2010,33(4)：30-31.
[123]王迪华,梁雪雯,周红.逍遥助卵煎对卵巢早衰患者血清性激素水平的影响.[J].中国妇幼保健,2007,22(33)：4687.
[124]任青玲,张昱,殷燕云.河车大造胶囊治疗卵巢早衰闭经35例.[J].药学与临床研究,2008,16(4)：297-299.
[125]段玮玮,夏阳.二仙汤加味治疗卵巢早衰42例临床观察.[J].吉林中医药,2008,28(3)：192-1 93.
[126]李颖,贾士奇,马仲丽.益肾养元丹治疗卵巢早衰120例临床观察.[J],陕西中医2008年29卷第3期,329
[127]杨丽,郭娜,许莉等.养血补肾方治疗卵巢早衰57例.[J]河北中医2009,31(3)：347-348.
[128]高慧,夏天.中药补肾调冲方治疗卵巢早衰的临床研究.[J].辽宁中医杂志,2007,34(11)：1557-1560.
[129]李霞.补肾调冲法治疗卵巢储备功能下降的临床研究.[D].2008硕士毕业论文：15-28.
[130]张雅萍,薄丽娟,劭丽娟.自拟补仙调经汤治疗卵巢功能早衰的临床研究.[J].中医药学报,2000,(1)：46-47.
[131]李宗恒,钟雪梅,代加莉.雌孕激素顺序法与补经汤治疗卵巢早衰的比较观察.[J].中国中医药科技,2002,9(3)：170-171.
[132]浦蕴星.针灸治疗闭经疗效分析.[J].上海针灸杂志,1994,13(6)：257-258.
[133]杨晓红,赖晓梅,黄祖波.针灸治疗卵巢早衰60例临床观察.[J].四川中医,2008,26(5)：106-107.
[134]沙桂娥,黄文川,马仁海,等.针灸治疗卵巢早衰不孕症84例的临床研究.[J].针灸临床杂志,1998,14(6)：16.
[135]何文扬.针刺治疗卵巢早衰症15例.[J].中国针灸,2000(7)：399.
[136]许学兵,刘红姣,表里经之俞募穴埋线治疗卵巢早衰33例.[J]中国民间疗法2008,(3)：12-13.
[137]陈瑞雪,李艳红,马晓晶.电针治疗卵巢早衰的疗效观察.[J]中国中医基础医学杂志,2009,5(3)：208.
[138]徐洁,蔡昱,谢群.卵巢早衰的中西医结合治疗与预防.[J].中国当代医学,2005,4(7)：83
[139]黄玉梅.中西医结合治疗卵巢早衰(45)例.[J].江西中医药,2008,39(2)：35.
[140]邢翠玲,杨玉荣,李爱芳.中西医结合治疗卵巢早衰症310例.[J].国医论坛,1995(51)：20.
[141]和平,张艳婷.中西医结合治疗卵巢功能早衰早期的临床探讨.[J].云南中医中药杂志,2004,25(2),3-4.
[142]钟雪梅,代嘉莉,李宗恒.补肾养经汤合人工周期法治疗卵巢早衰38例临床观察.[J].江苏中医药,2005,26(8)：21-22.
[143]赵建春,徐萍,宋亚光.中西医结合治疗卵巢早衰40例观察.[J].四川中医,2004,22(7)：69-70.
[144]衣忠玲.卵巢早衰治疗体会.[J].实用中医药杂志,2000,16(9)：38.
[2]Laml T, Schulz-Lobmeyr I, Obruca A,et al. Premature ovarian failure:etiology and prospects[J].Gynecol Endocrinol,2000,Aug; 14(4):292-302.
[3]Kalantaridou SN, Davis SR, Nelson LM. Premature ovarian failure[J].Endocrinol Metab Clin North Am,1998,Dec;27(4):989-1006.
[4]Santoro N. Mechanisms of premature ovarian failure. [J].An Endocrinol 2003,64:87-92.
[5]Timmreck LS, Reindollar RH:Contemporary issues in primary amenorrhea. [J].Obstetgynecol Clin North Am 2003,30:287-302.
[6]Coulam CB, Adamson SC, Annegers JF.Incidence of premature ovarian failure [J].Obstet Gynecol,1986,Apr;67(4):604-6.
[7]司徒仪主编.中西医结合妇产科学[M].北京：北京科学技术出版社,2003：155.
[8]SHYR-Yeu Lin, John R Morrison, David J, et al. Regulation of ovarian function by the TGF-superfamily and follistatin [J].Reproduction,2003,126:133-148.
[9]Welt. Female reproductive aging ismarked by decreased secretion of dimeric inhibin[J]. Clin EndocrinolMetab,1999,84(1):105
[10]La Marca A, Stabile G, Tirelli A, et al. Anti-mullerian hormonemeasurement on any day of the menstrual cycle strongly predicts o-varian response in assisted reproductive technology [J]. Hum Re-prod,2007,22(3):766-771.
[11]Andersen CY,ByskovAG.Estradiol and regulation ofantiMullerian hormone, inhibin-A, and inhibin-B secretion:analysis of small antral and preovulatory human folliclesfluid[J]. J Clin EndocrinolMetab,2006,91:4064-4069
[12]KevenaarME,Themmen AP,Laven JS, et a.l Anti-Mu-llerian hormone and anti-Mullerian hormone type 11 receptor polymorphisms are associatedwith follicularphase estradiol levels in normo-ovulatory women [J]. Hum Reprod,2007,22:1547-1554
[13]张敏慧 丛晶者等.抗苗勒管激素与卵巢功能[J]国际生殖健康／计划生育杂志,2009,28(5)：281-286.
[14]Bussolino F,Mantovani A,Persico G. Molecular mechanisms of blood vessel formation[J]. TIBS,1997,221251
[15]GleieherN,WeglloferA, Barad DH.A Pilot study of Premature ovarian seneseenee: Ⅱ.Different genotype and Pllenotype for genetic and autoimmune etiologies.FertilSteri, Ⅰ 2008,3(7):203-205.
[16]DongL,JiangL,MenW, et al.Preventive and therapeutic effects of BushenHuoxue Recipe on autoimmune premature ovarian failure in mice.ZhongxiyiJieHexueBao,2008,6(3):294-297.
[17]Gregoraszczuk EL,Slomczynska M. Beta-endorphin inhibition ofprogesterone secretion by porcine granulosa cells during follicledevelopment[J].Reprod NutrDev,1998,38(3):227-234.
[18]Besedovsky H, Sorkin E. Network of immune-neuroendocrine interactions[J].Clin Exp Immunol,1977,Jan; 27(1):1-12.
[19]陈晓萍,徐远扬.神经、内分泌、免逸网络的通用生物学语言[J].自然杂志,2002,24(4)：194.
[20]郭蕾,王永炎,张志斌.关于证候概念的诊释[J1.北京中医药大学学报,2003,26(2)：5-8.
[21]张焱,回世洋,张庆荣.Z2医五脏调控系统与神经内分泌免逸调节网络的关系[J].中华中医药学刊,2009,27(1)：188-189.
[22]金婧,阮祥燕.卵巢早衰的病因研究进展.[J],实用妇产科杂志,2007,23(3)：142-145.
[23]Kauffman RP, Castracane VD.Premature ovarian failure asociated with autoimmune polyglandular syndrome:pathophysiological mechanisms and future fertility. [J].Women's Health(Larchmt),2003,12(5):513-520.
[24]杨业洲.卵巢早衰.[M].北京：人民卫生出版社,2004,102-108.
[25]孙晓溪,李大金,王明雁,等.自身免疫性卵巢功能损伤与TH1/TH2型细胞因子表达关系的研究.[J].中国免疫学杂志,2001,17(2)：96-98.
[26]Chernyshov VP, RadyshTV, Gura Ⅳ, et al.Immune disorders in women with premature ovarian failure in initial period. [J].Am J Reprod Immunol.2001,46(3):220-225.
[27]王一峰,欧汝强,杨宁,等.卵巢早衰患者血消抗透明带抗体和肿瘤坏死因子-α,γ-干扰素及白细胞介素-2的分析.[J].生殖医学杂志,2002,11(1)：7-10.
[28]Monnier Barbarino P, Forges T, Faureg C.gonadal antibodies interfeting with female reproduction. [J].Best-Pract-Res-Clin-Endocrinol-Metab.2005,19(1):135-148.
[29]DoldiN, Belvisi L, Bassan M, et al.Premature ovarian failure:steroid synthesis and autoimmunity. [J].gynecol Endocrinol.1998,12(1):23-28.
[30]赵建英,高晓秋,史樱.35例卵巢早衰患者雌二醇与睾酮水平分析.[J].生殖医学杂志,2000,9(6)：355-356.
[31]Bakalov VK, Vanderhoof VH, Bondy CA, et al. Adrenal antibodies detect asymptomatic auto-immune adrenal insufficiency in young women with spontaneous premature ovarian failure. [J].Hum Reprod,2002,17(8):2096-2100.
[32]李珂,补肾健脾调肝法治疗卵巢早衰及对POF模型大鼠血清INHB、VEGF的影响.广州中医药大学2006届博士学位论文,2006,7.
[33]杨桂艳,王一理,曹绩孙等.卵巢早衰患者外周血CD4+、THI/TH2细胞分布研究.生殖医学杂志,1998；7(4)：227—230.
[34]Arif S, Underhill JA, Donaldson P, et al. Human leukocyte antigen-DQB1*genotypes aspartate at position 57 are associated with 3β-Hydroxysteroid dehydrogenase auto-immunity in premature ovarian failure. [J].Clin Endocrinol Metab,1999,84(3): 1056-1060.
[35]Reimand K, Peterson P, Hyoty H.3beta-hydroxysteroid dehydrogenase autoantibodies are rare in premature ovarian failure. [J].Clin Endocrinol Metab.2000 Jun; 85(6):2324-6.
[36]Zhi-Bin Tong, Lawrence M. Nelson. A Mousegene Encoding an Oocyte Antigen Associated with Autoimmune Premature Ovarian Failure. [J]. Endocrinology.1999, 140(8):3720-3726.
[37]Rhim SH, Millar SE, RobeyF, et al. Autoimmune disease of the ovary induced by a ZP3 peptide from the mouse zona pellucid. [J]. Clin Invest.1992 Jan,89(1):28-35.
[38]Naz RK, Thurston D, Santoro N. Circulating tumor necrosis factor (TNF)-alpha in normally cycling women and patients with premature ovarian failure and polycystic ovaries. [J]. Am J Reprod Immunol.1995 Sep; 34(3):170-5.
[39]Giovannigrasso, Michela Muscettola. Possible Role of Interferon-Gamma in Ovarian Function [M]. New York:Annals of the New York Academy of Sciences,1992:650(15): 191-196.
[40]Russell P. The clinicopathological features of primary ovarian failure. [J]. Verh Dtschges Pathol.1997,81:197-209.
[41]谢端筱,欧汝强,杨宁等.卵巢早衰与细胞因子TNF-a, IFN-y和IL-2的相关性研究.[J].广东医学,1999,20(11)：831-832.
[42]Felice Petraglia, Beda Hartmann, Stefano Luisi, et al. Low levels of serum inhibin A and inhibin B in women with hypergonadotropic amenorrhea and evidence of high levels of activin A in women with hypothalamic amenorrhea. [J]. Fertility and Sterility,1998, 70(5):907-912.
[43]陈蔚,雷小敏.卵巢早衰病因学研究进展.[J].生殖与避孕,2004,24(1)：43-7.
[44]李洁,杨菁.卵巢早衰动物模型建立的研究进展.[J].生殖与避孕,2006,26(9)：550-53.
[45]Yang PM, Chen HC, Tsai JS, et al. Cadmium induces Ca2+-dependent necrotic cell death through calpain-triggered mitochondrial depolarization and reactive oxygen species-mediated inhibition of nuclear factor-kappaB activity. [J]. Chem Res Toxicol. 2007,20(3):406-15.
[46]Sharabidze N, Burkadzeg, Sabakhtarashvili M. Cell adhesion and apoptosis in ovarian stromal hyperplasia and hyperthecosis. [N].georgian Med News.2006, (131):33-7.
[47]Depalo R, Nappi L, Loverrog, et al. Evidence of apoptosis in human primordial and primary follicles. [J]. Hum Reprod.2003,18(12):2678-82.
[48]王雪峰,何援利.凋亡调节基因bcl-2/bax与卵巢早衰的关系.[J],生殖与避孕,2008,28(8)：487-90.
[49]桑海滨.卵巢早衰的中西医诊治.贵阳中医学院学报,2006,28(4)：41-43.
[50]Morita Y, Perezgl, Paris F, et al. Oocyte apoptosis is suppressed by disruption of the acid sphingomyelinasegene or by sphingosine-1-phosphate therapy. [J].Nat Med.2000, 6(10):1109-14.
[51]Chauhan P, Sodhi A, Tarang S. Cisplatin-treated murine peritoneal macrophages induce apoptosis in L929 cells:role of Fas-Fas ligand and tumor necrosis factor-tumor necrosis factor receptor 1. [J]. Anticancer Drugs.2007,18(2):187-96.
[52]艾红,胡海燕,曹攒孙.自然排卵周期中子宫卵巢血流与性激素、血管内皮生长因子的关系.[J].中国医学影像杂志,2004,12(4)：286-88.
[53]Agrawal R, ConwaygS, Sladkevicius, et al. Serum vascular endothelialgrowth factor (VEGF) in the normal menstrual cycle:association with changes in ovarian and uterine Doppler blood flow. [J]. Clin Endocrinol (Oxf).1999,50(1):101-06.
[54]Seang Lin Tan, Jamal Zaidi, Stuart Campbell, et al. Blood flow changes in the ovarian and uterine arteries during the normal menstrual cycle. [J]. American Journal of Obstetrics &gynecology.1996,175(3):625-31.
[55]van Kasteren YM, Hundscheid RD, Smits AP, et al. Familial idiopathic premature ovarian failure:an overrated and underestimatedgenetic disease?. [J]. Hum Reprod.1999, 14(10):2455-59.
[56]Vegetti W, Marozzi A, Manfredini E, et al. Premature ovarian failure. [J]. Mol Cell Endocrinol.2000,161(1-2):53-7.
[57]Progetto Menopausa Italia Studygroup. Premature ovarian failure:frequency and risk factors among women attending a network of menopause clinics in Italy. [R]. BJOG.2003, 110(1):59-63.
[58]Testag, Chiaffarino F, Vegetti W, et al. Case-control study on risk factors for premature ovarian failure. [J].gynecol Obstet Invest.2001,51(1):40-3.
[59]Frisch RE. Body fat, menarche, fitness and fertility. [J]. Hum Reprod.1987,2(6): 521-33.
[60]Harlow BL, Signorello LB. Factors associated with early menopause. [J]. Maturitas, 2003,5(1):3.
[61]徐苓,宋亦军.卵巢早衰的临床表现和诊断标准.[J].实用妇产科杂志,2003,19(4)：195-196.
[62]张凇文,王军华,周红等.北京地区女性绝经期相关年龄相关因素的调查分析[C].第一次绝经相关问题研讨会论文汇编,2001,11(4)：164.
[63]许小凤,范春,包广勤等.卵巢早衰与卵巢储备功能下降的相关发病因素探讨[J].2011,11(4)：400-403.
[64]Luborsky JL, Meyer P, Sowers MF, et al. Premature menopause in a multi-ethnic population study of the menopause transition. [J]. Hum Reprod,2003,18(1):199-206.
[65]Kalantaridou SN, Nelson LM. Premature ovarian failure is not premature menopause. [M]. Ann N Y Acad Sci.2000,900:393-402.
[66]Ataya KM, McKanna JA, Weintraub AM, et al. A luteinizing hormone-releasing hormone agonist for the prevention of chemotherapy-induced ovarian follicular loss in rats. [J]. Cancer Res.1985 Aug; 45(8):3651-6.
[67]付霞霏,何援利.化疗所致卵巢早衰动物模型的建立.[J]广东医学,2008,29(12)：1952-1954.
[68]Yu X, Kamijima M, Ichiharag, et al.2-Bromopropane causes ovarian dysfunction by damaging primordial follicles and their oocytes in female rats. [J]. Toxicol Appl Pharmacol. 1999 Sep 15; 159(3):185-93.
[69]艾浩,牛建昭,薛晓等.顺铂致小鼠卵巢功能早衰肝肾阴虚证机制研究,[J]北京中医药大学学报,2006,29(6)：401-403.
[70]高慧,杨涓,韩冰,等.药物型卵巢早衰动物模型的研究.[J]国医论坛,2007,22(3)：22-26.
[71]付莉,赵怡璇,李守柔.卵巢早衰实验动物模型的建立[J].生殖医学杂志,2006,15(3)：179-183.
[72]林建华,严隽鸿,林其德.自身免疫性卵巢衰退小鼠动物模型的建立[J].上海医学,1999,12(12)：747-748.
[73]Bagavant H, Adams S, Terranova P, et al. Autoimmune ovarian inflammation triggered by pro inflammatory (Th1) T cells is compatible with normal ovarian function in mice. [J]. Biol Reprod.1999 Sep; 61(3):635-42.
[74]司卉良,陈友香,侯安继,等.补肾汤对激素致更年期肾阳虚证大鼠卵巢功能的影响.[J].中药药理与临床,2005,21(2)：44-45.
[75]Laissue P, Christin-Maitre S, Touraine P, et al. Mutations and sequence variants ingDF9 and BMP 15 in patients with premature ovarian failure. [J]. Eur J Endocrinol.2006 May; 154(5):739-44.
[76]Kelkar RL, Meherji PK, Kadam SS, et al. Circulating auto-antibodies against the zona pellucida and thyroid microsomal antigen in women with premature ovarian failure. [J]. J Reprod Immunol.2005 Jun; 66(1):53-67.
[77]Omoto Y, Lathe R, Warner M, et al. Early onset of puberty and early ovarian failure in CYP7B1 knockout mice. [J]. Proc Nat1 Acad Sci U S A.2005 Feb 22; 102(8):2814-9. Epub 2005 Feb 14.
[78]Takai Y, Canning J, PerezgI, et al. Bax, caspase-2, and caspase-3 are required for ovarian follicle loss caused by 4-vinylcyclohexene diepoxide exposure of female mice in vivo. [J]. Endocrinology.2003 Jan; 144(1):69-74.
[79]Yu WJ, Lee BJ, Nam SY, et al. Modulating effects of Koreanginseng saponins on ovarian function in immature rats. [J]. Biol Pharm Bull.2003 Nov; 26(11):1574-80.
[80]Olff M. Stress, depression and immunity:the role of defense and coping styles. [J]. Psychiatry Res.1999 Jan 18,85(1):7-15.
[81]连晓媛,丁岩,陈奇,等.重复制动应激对雌性大鼠卵巢功能的影响.[J].中药新药与临床药理,2004,15(6)：373-376.
[82]Bandyopadhyay S, Chakrabarti J, Banerjee S, et al. Prenatal exposure to highgalactose adversely affects initialgonadal pool ofgerm cells in rats. [J]. Hum Reprod.2003 Feb, 18(2):276-82.
[83]李洁,杨菁,陈嫒,等.利用不同浓度D(+)半乳糖建立卵巢早衰动物模型.[J].武汉大学学报(医学版),2006,27(5)：614-619.
[84]莫中福,殷秀玲,尹青,等DahlS高血压大鼠卵巢细胞凋亡相关因素的实验研究.[J].生殖与避孕,2002,22(5)：263-265.
[85]张晓金,归绥琪,徐丛剑.补肾复方调控更年期大鼠卵巢功能的实验研究.[J].现代中西医结合杂志,2003,12(19)：2044-2047.
[86]张绍芬,谢倩,哈灵侠,等.戊酸雌二醇对更年期大鼠脾细胞凋亡及卵巢功能的作用.[J].生殖医学杂志,2006,15(6)：388-392.
[87]蔡少锋,康幼雯.中西医结合人工周期疗法抗卵巢早衰的临床探讨.[J].中国民间疗法,1 997(6)：20.
[88]王锡珍,赖毛华.中医对卵巢早衰的认识和证治体会.[J]浙江中医学院学报,2003,27(5)：21-22
[89]王菊素.田淑霄教授对卵巢早衰与肾虚血瘀相关性的见解及治疗经验.[J].陕西中医,2003,11(24)：1015.
[90]朱玲,.罗颂平.卵巢早衰的病因病机及其论治.[J].中医药学刊,2003,21(1)：143-144.
[91]夏天.卵巢早衰的中西医结合研究进展.[J].辽宁中医杂志,2004,31(8)：707-708.
[92]黄健萍.卵巢早衰的中医治疗述要.[J].辽宁中医学院学报,2004,6(2)：126.
[93]张玉珍,史云.试论中医药防治卵巢早衰的思路与方法.[J]中医杂志,2005,46,(增刊)：116-117.
[94]魏智慧,邓琳文.浅议从肾论治卵巢早衰.[J].甘肃中医,2007,20(9)：39-40.
[95]刘霞,谢萍.浅析卵巢早衰的病机和证治.[J].江西中医学院学报,2007,19(4)：18-19.
[96]高慧,杨涓,李巧芬.中医学对卵巢早衰病因病机的认识.[J].甘肃中医,2007,20(8)：47-48.
[97]蔡立荣,李大金,孙晓溪,等.补肾活血方对小鼠实验性卵巢早衰防治作用的研究.[J].中国中西医结合杂志,2001,21(2)：126.
[98]钟雪梅,周灵,杨家林.补经合剂促卵泡发育和排卵的实验研究.江苏中医,1998,19(3)：45-46.
[99]董莉,谈嫒,平瑜佳等.不同治法方药治疗小鼠自身免疫性卵巢功能衰退的比较研究.[J]中西医结合学报2010,8(1)：86-89.
[100]张军武.归肾丸对大鼠卵巢早衰防治作用的实验研究.[J]中国现代医药杂志2008,10(6)：79-80.
[101]孙永忠,程泾,石其贤.滋肾益冲抗衰汤治疗卵巢早衰实验研究.[J]实用中医药杂志,2008,24(2)：71-73.
[102]朱玲,罗颂平,许丽绵等.左归丸对免疫性卵巢早衰小鼠抗卵巢抗体的影响.现代中西医结合杂志,[J]2006,15(4)：435-438.
[103]朱玲,罗颂平,许丽绵等.左归丸对免疫性卵巢早衰小鼠卵巢Fas、Fas-L表达的影响,[J]江西中医学院学报2008,20(1)：52-55.
[104]杨兰英.六味地黄丸治疗卵巢早衰的实验研究.[J].山西中医学院学报,2002,3(1)：21
[105]梁欣韫,孙晓芳,牛建昭等.育泡饮对自身免疫性卵巢损伤小鼠卵巢颗粒细胞的超微结构影响.[J]中华中医药学刊,2008,26(3)：502-504.
[106]李军,王翠霞,陈莹等.补肾复方延缓雌性大鼠卵巢早衰的实验研究.[J]辽宁中医杂志2007,3(5)：681-682.
[107]高慧,杨涓,夏天等.补肾调冲方对卵巢早衰大鼠卵巢相关细胞因子TNF-α、IFN-γ蛋白和基因表达的影响.[J]国医论坛2008,23(2)：32-39.
[108]李沛霖,侯莉莉,杜永红.补肾调冲含药血清对大鼠卵巢细胞体外分泌功能的影响.[J].时珍国医国药,2006,17(3)：354-355.
[109]滕秀香.122例卵巢早衰患者中医证候分析及致病因素调查[J] Chinese Journal of Information on TCM,2008,15(4):18-20.
[110]张朝辉,张亚辉.阳和汤治疗卵巢早衰症治验.[J].北京中医,2001,17(4)：16
[111]陈秀芳.辨证治疗卵巢早衰22例.[J].四川中医,2000,18(1)：35
[112]梁雪雯,苏建芬,王迪华,等.逍遥助卵煎治疗卵巢早衰30例.[J].上海中医药杂志,2006,40(2)：31-32.
[113]许昕,藤秀香,佟庆.补肾养血调经法治疗有卵泡型卵巢早衰.[J].北京中医,2001,(4)：21
[114]魏丹,肖秋霞.补肾养血调经法治疗卵巢早衰12例.[J].江西中医药,2008,39(30)：43-44.
[115]种丽群.补肾活血化淤法治疗卵巢早衰24例.[J].实用中医药杂志,2006,22(11)：675。
[116]兰英,魏巍.柴瑞霭治疗卵巢早衰验案举隅.[J].山西中医,2001,17(4)：5
[117]钱菁.夏桂成妇科病症验案举隅.[J].辽宁中医杂志,2005,32(11)：1198-1199.
[118]李云君,姜惠中.姜惠中治疗卵巢早衰的经验.[J].湖北中医杂志,2006,28(6)：28-29.
[119]周华,赵春梅.中西医治疗卵巢早衰临床研究述略.[J].中医药学刊,2004,22<5)：903.
[120]秦薇,周梅玲,陈祥云.中药人工周期疗法治疗卵巢功能早衰45例.[J].国医论坛,2004,19(1)：30-33.
[121]池雷,夏桂成.夏桂成教授调周疗法治疗月经病述要.实用医技杂志,2008,15(33)：4914.
[122]武淑慧,庆璐,崔晓萍.崔晓萍教授循期阴阳序贯疗法治疗卵巢早衰.[J]陕西中医学院学报2010,33(4)：30-31.
[123]王迪华,梁雪雯,周红.逍遥助卵煎对卵巢早衰患者血清性激素水平的影响.[J].中国妇幼保健,2007,22(33)：4687.
[124]任青玲,张昱,殷燕云.河车大造胶囊治疗卵巢早衰闭经35例.[J].药学与临床研究,2008,16(4)：297-299.
[125]段玮玮,夏阳.二仙汤加味治疗卵巢早衰42例临床观察.[J].吉林中医药,2008,28(3)：192-1 93.
[126]李颖,贾士奇,马仲丽.益肾养元丹治疗卵巢早衰120例临床观察.[J],陕西中医2008年29卷第3期,329
[127]杨丽,郭娜,许莉等.养血补肾方治疗卵巢早衰57例.[J]河北中医2009,31(3)：347-348.
[128]高慧,夏天.中药补肾调冲方治疗卵巢早衰的临床研究.[J].辽宁中医杂志,2007,34(11)：1557-1560.
[129]李霞.补肾调冲法治疗卵巢储备功能下降的临床研究.[D].2008硕士毕业论文：15-28.
[130]张雅萍,薄丽娟,劭丽娟.自拟补仙调经汤治疗卵巢功能早衰的临床研究.[J].中医药学报,2000,(1)：46-47.
[131]李宗恒,钟雪梅,代加莉.雌孕激素顺序法与补经汤治疗卵巢早衰的比较观察.[J].中国中医药科技,2002,9(3)：170-171.
[132]浦蕴星.针灸治疗闭经疗效分析.[J].上海针灸杂志,1994,13(6)：257-258.
[133]杨晓红,赖晓梅,黄祖波.针灸治疗卵巢早衰60例临床观察.[J].四川中医,2008,26(5)：106-107.
[134]沙桂娥,黄文川,马仁海,等.针灸治疗卵巢早衰不孕症84例的临床研究.[J].针灸临床杂志,1998,14(6)：16.
[135]何文扬.针刺治疗卵巢早衰症15例.[J].中国针灸,2000(7)：399.
[136]许学兵,刘红姣,表里经之俞募穴埋线治疗卵巢早衰33例.[J]中国民间疗法2008,(3)：12-13.
[137]陈瑞雪,李艳红,马晓晶.电针治疗卵巢早衰的疗效观察.[J]中国中医基础医学杂志,2009,5(3)：208.
[138]徐洁,蔡昱,谢群.卵巢早衰的中西医结合治疗与预防.[J].中国当代医学,2005,4(7)：83
[139]黄玉梅.中西医结合治疗卵巢早衰(45)例.[J].江西中医药,2008,39(2)：35.
[140]邢翠玲,杨玉荣,李爱芳.中西医结合治疗卵巢早衰症310例.[J].国医论坛,1995(51)：20.
[141]和平,张艳婷.中西医结合治疗卵巢功能早衰早期的临床探讨.[J].云南中医中药杂志,2004,25(2),3-4.
[142]钟雪梅,代嘉莉,李宗恒.补肾养经汤合人工周期法治疗卵巢早衰38例临床观察.[J].江苏中医药,2005,26(8)：21-22.
[143]赵建春,徐萍,宋亚光.中西医结合治疗卵巢早衰40例观察.[J].四川中医,2004,22(7)：69-70.
[144]衣忠玲.卵巢早衰治疗体会.[J].实用中医药杂志,2000,16(9)：38.