摘要
第一部分:血管紧张素Ⅱ对肺微血管内皮细胞Apelin-APJ mRNA表达的影响
目的研究血管紧张素Ⅱ(AngⅡ)对肺微血管内皮细胞Apelin-APJ mRNA表达的影响。
方法培养新生鼠肺微血管内皮细胞(PMVECs),利用RT-PCR检测PMVECsApelin-APJmRNA表达。取2-4代细胞,以5×10~5个/孔密度接种至6孔板,培养至80%融合后,进行药物干预实验:①加AngⅡ入6孔板中,至终浓度分别为0、10~(-9)、10~(-8)、10~(-7)、10~(-6)M(mol/L),每组设立4个复孔,继续培养24h终止实验,检测不同浓度AngⅡ对PMVECs Apelin-APJmRNA表达的影响;②加AngⅡ入6孔板至终浓度为10~(-7)M,分别在0h、1h、6h、12h、24h、48h时间点终止实验。同时在0h、24h、48h点设立空白对照,加入等体积PBS。每组4个复孔。检测不同时间点10~(-7)M AngⅡ对PMVECs Apelin-APJmRNA表达。
结果①PMVECs存在Apelin-APJmRNA表达。②与对照组(OM AngⅡ)比较,10~(-9)M AngⅡ作用24h后Apelin mRNA表达上调(P＜0.01),随着AngⅡ浓度上升,Apelin mRNA表达呈浓度依赖性持续下调(P＜0.05或P＜0.01);不同浓度AngⅡ(10~(-9)-10~(-6)M)呈浓度依赖性导致APJmRNA表达下降(P＜0.05或P＜0.01)。③10~(-7)MAngⅡ作用于PMVECs后,Apelin mRNA表达在6h内增加(P＜0.05),其中在1h达到高峰(P＜0.01),6h后Apelin mRNA表达随时间延长明显降低(P＜0.01);APJ mRNA表达在0h-12h内有增加趋势,12h后APJ mRNA表达随10~(-7)M AngⅡ作用时间延长明显下调(P＜0.01);0h、24h、48h时间点空白对照Apelin-APJmRNA表达无明显改变(P＞0.05)。
结论AngⅡ对PMVECs Apelin-APJ基因表达水平有明显影响,Apelin-APJ系统可能参与AngⅡ对内皮细胞损伤机制。
第二部分:Apelin对肺血管内皮细胞增殖和凋亡的影响
目的探讨Apelin对肺血管内皮细胞增殖与凋亡的影响。
方法培养新生大鼠肺微血管内皮细胞(PMVECs),取2-4代PMVECs用于实验:①以2000个/孔的密度接种至96孔板,分为3组:对照组加入等容量PBS;低浓度组(10~(-8)M Apelin组)加入10~(-6)M Apelin;高浓度组(10~(-6)M Apelin组)加入10~(-6)MApelin。每组6个复孔。48h后用四甲基偶氮唑蓝比色法(MTT法)检测各组OD值,计算细胞增殖率。②以5×10~5传代至25ml的培养瓶,分为5组:对照组加入等容量PBS;低浓度组(10~(-8)M Apelin组)加入10~(-8)M Apelin;高浓度组(10~(-6)M Apelin组)加入10~(-6)M Apelin;AngⅡ组加入10~(-7)M AngⅡ;AngⅡ+Apelin组加入10~(-8)MApelin,10min后加入10~(-7)M AngⅡ。每组5个复孔。24h后用流式细胞仪检测细胞凋亡率。
结果①与对照组比较,低浓度Apelin和高浓度Apelin都可导致PMVECs OD值增加,增殖率增加2.5倍以上(P＜0.01)。低浓度组和高浓度组之间没有统计学差异。②与对照组比较,低浓度Apelin组和高浓度Apelin组的PMVECs凋亡率降低50%以上(P＜0.05),AngⅡ组PMVECs凋亡率明显升高(P＜0.01);与AngⅡ组比较,AngⅡ+Apelin组PMVECs凋亡率明显降低(P＜0.01)。
结论Apelin促进体外培养大鼠PMVECs增殖,抑制PMVECs凋亡。
第三部分:Apelin对肺微血管内皮细胞保护作用及其释放NO的机制研究
目的探讨Apelin对血管紧张素Ⅱ(AngⅡ)诱导肺血管内皮细胞释放ET-1的影响和刺激一氧化氮(NO)的释放机制。
方法培养新生鼠肺微血管内皮细胞(PMVECs),取2-4代PMVECs用于实验。
①Apelin在不同时间点对PMVECs释放NO的影响:以1×10~5个/孔密度接种至24孔板,80%融合后,无血清培养24h,分为2组:Apelin组,加入Apelin至终浓度为10~(-8)M;对照组,加入等量无血清DMEM培养液。每组设立4个复孔。继续培养,并分别在0min、2min、5min、15min、30min五个时间点取培养液测NO浓度。
②Apelin对AngⅡ诱导PMVECs释放ET-1的影响:分为4组:AngⅡ组,加入AngⅡ至终浓度为10~(-7)M;AngⅡ+10~(-8)M Apelin组,加入10~(-7)M AngⅡ前5min加入10~(-8)M Apelin预处理;AngⅡ+10~(-6)M Apelin组,加入10~(-7)M AngⅡ前5min加入10~(-6)M Apelin预处理;对照组,加入等量无血清DMEM培液。每组设立4个复孔。继续培养,并分别在0min、30min、6h、24h四个时间点取培养液检测ET-1浓度。
③Apelin对PMVECs Akt/eNOS磷酸化影响:以5×10~5个/孔密度接种至6孔板,80%融合后,无血清培养24h,分为3组:Apelin组,加入Apelin至终浓度为10~(-8)M;Apelin+Akt inhibitor组,加入Apein前用5uM浓度的Akt抑制剂预处理30min;对照组,加入等量DMEM培液。每组设立3复孔。继续培养5min后立即提取总蛋白,用于Western Bolt检测磷酸化Akt和磷酸化eNOS表达。
结果
①Apelin在不同时间点对PMVECs释放NO的影响:
在PMVECs培养液中加入10~(-8)M的Apelin后,与对照组比较,培养液中NO浓度在2min、5min、15min时间点都出现增加(P＜0.01或P＜0.05);其中在5min时间点达到高峰(与对照组比较,P＜0.01);在30min时间点,NO浓度与对照组比较无统计学差异。
②Apelin对AngⅡ诱导PMVECs释放ET-1的影响:
与对照组比较,AngⅡ组培养液中30min、6h、24h时间点的ET-1浓度都明显升高(P＜0.05)。与AngⅡ组比较,AngⅡ+10~(-8)M Apelin组培养液中30min、6h、24h时间点的ET-1浓度都明显降低(P＜0.05),接近对照组。与AngⅡ组比较,AngⅡ+10~(-8) Apelin组的ET-1浓度有降低趋势,但无统计学意义。
③Apelin对PMVECs Akt/eNOS磷酸化影响:
与对照组比较,Apelin组PMVECs Akt和eNOS磷酸化表达明显增加(P＜0.01)。与Apelin组比较,Apelin+Akt inhibitor组PMVECs eNOS磷酸化表达明显降低(P＜0.01)。
结论Apelin能促进PMVECs释放NO,抑制AngⅡ诱导的ET-1释放。Akt/eNOS磷酸化增加是Apelin促进NO释放机制之一,并可能参与细胞保护。
PartⅠ:The effect of angiotensinⅡon expression of Apelin-APJ mRNA in pulmonary microvascular endothelial cells
Objective Exploring the effect of angiotensinⅡ(AngⅡ) on expression of Apelin-APJ mRNA in pulmonary microvascular endothelial cells(PMVECs).
Methods Pulmonary microvascular endothelial cells from neonatal rat of 24h after birth were cultured.Expressions of Apelin-APJ mRNA in endothelial cells were determined by reverse transcription-polymerase chain reaction(RT-PCR).PMVECs were used between passages 2 and 4.5×10~5 cells grown in 6-well plates at subconfluence were stimulated with various concentrations of AngⅡ(0、10~(-9)、10~(-8)、10~(-7)、10~(-6)M) for analyzing apelin and APJ mRNA expressions by RT-PCR at 24hr and with AngⅡof 10~(-7) M for apelin and APJ mRNA expression at different time points of 0hr,1hr,6hr, 12hr,24hr,48hr.At the same time analyzing their expressions in blank groups at three time points of 0hr,24hr and 48hr without stimulation of AngⅡ.every experience was repeated for 4 times.
Results①PMVECs expressed apelin and APJ mRNA determined by RT-PCR.②compared with control group(OM AngⅡ),10~(-9) M AngⅡinduced upregulation of apelin mRNA expressions in PMVECs at 24hr(P＜0.01), higher concentrations of AngⅡ(10~(-8)、10~(-7)、10~(-6)M) induced concentrationdependent downregulation of apelin mRNA expressions(P＜0.05 or P＜0.01); different concentrations of AngⅡ(10~(-9)、10~(-8)、10~(-7)、10~(-6)M) concentrationdependently downregulated expressions of APJ mRNA.③10~(-7) M AngⅡupregulated apelin mRNA expressions in short time(within 6hr,P＜0.05) with a peak at 1hr(P＜0.01),and markedly downregulated them 6 hrs later (P＜0.01);10~(-7)M AnglI also markedly time-dependently downregulated APJ mRNA expressions after 12 hrs(P＜0.01);apelin and APJ mRNA expressions did not significantly change in PMVECs without treatment with AngⅡat 3 time points of 0hr,24hr and 48hr.
Conclusion AngⅡcan markedly effect on apelin and APJ mRNA expressions in PMVECs,and apelin-APJ system may involve in the mechanism of injury of AngⅡon endothelial cells.
PartⅡ:Effects of Apelin on proliferation and apoptosis in pulmonary vascular endothelial cells
objective To investigate the effects of apelin on proliferation and apoptosis in pulmonary vascular endothelial cells.
Methods Pulmonary microvascular endothelial cells(PMVECs) between passage 2 and 4 cultured from neonatal rat of 24 hours after birth were starved serum-free for 24hours,and then were used to following studies:①2×10~3 cells per well grown in 96-well plate were divided randomly into 3 groups:Control group(C group) was added with equal volume of PBS;Lower concentration group(10~(-8) M group) with 10~(-8) M of apelin; Higher concentration group(10~(-6) M group) with 10~(-6) M of apelin.MTT assay was used to assess cell proliferation rates after cells were continuously incubated for 48 hours.②5×10~5 cells grown in cultured-bottles of 25 ml were divided into 5 groups:Control group were added with equal volume of PBS;Lower concentration group(10~(-8) M group) with 10~(-8) M of apelin; Higher concentration group(10~(-6) M group) with 10~(-6) M of apelin; AngiotensinⅡ(AngⅡ) group with 10~(-7) M of AngⅡ;AngⅡ+Apelin group with 10~(-7) M of AngⅡand 10~(-8)M of apelin.The apoptosis rates of PMVECs in each group were detected by flow cytometry after the cells were incubated for 24 hours.
Results①The proliferation rates of the PMVECs significantly increased in both lower and higher concentration of apelin groups compared to control group(P＜0.01),and there were not significantly different between lower and higher concentration of apelin groups.②The apoptosis rates of the PMVECs significantly decreased in both lower and higher concentration groups but dramatically increased in AngⅡgroup,compared to control group(P＜0.05 or P＜0.01);The apoptosis rates were lower in AngⅡ+Apelin group than in AngⅡgroup(P＜0.01).
Conclusion Apelin can promote proliferation of PMVECs and inhibit apoptosis.
PartⅢ:Protective effects of Apelin on pulmonary microvascular endothelial cells and the mechanism of it' s promoting NO release
Objective To investigate the effects of Apelin on release of ET-1 induced by AngiontensinⅡand it' s mechanism of stimulating NO release in pulmonary microvascular endothelial cells(PMVECs).
Methods PMVECs between passage 2 and 4 cultured from neonatal rat of 24 hours after birth were starved serum-free for 24 hours,and then used to following studies.
①The effects of Apelin on release of NO in PMVECs at different times: 1×10~5 cells per well grown in 24-well plate were divided randomly into 2 groups:Apelin group was added with 10~(-8) M Apelin;control goup with equal volume of serum-free DMEM.Then concentration of NO in cultural medium were detected in both groups at five different time points of 0min,2min,5min,15min and 30min.
②The effects of Apelin on ET-1 release in PMVECs induced by AngiotensinⅡ:PMVECs grown in 24-well plats were randomly divided into 4 groups: AngⅡgroup was added with 10~(-7)M Apelin;AngⅡ+10~(-8)M Apelin group incubated with 10~(-8)M Apelin at 5 min prior to stimulation of 10~(-7)M AngⅡ; AngⅡ+10~(-6) M Apelin group with 10~(-6)M Apelin 5min prior to 10~(-7)M AngⅡ; control group with equal volume of serum-free DMEM.ET-1 concentrations in cultural medium were measured in all groups at different time points of 30min,6hr and 24hr.
③The effects of Apelin on phosphorylations of endothelial nitric-oxide synthase and Akt:5×10~5 cells per well grown in 6-well plates were divided into 3 groups:Apelin group was added with 10~(-8)M of Apelin; Apelin+Akt inhibitor group was pretreated with 5uM of Akt inhibitor 30min prior to incubated with Apelin;control group with equal volume of serum-free DMEM.5min later western blot was used to assay protein expressions of phosphorylations of endothelial nitric-oxide synthase and Akt.
Results
①The effects of Apelin on release of NO in PMVECs at different times: compared with control group,concentrations of NO in cultural medium increased at the three time points of 2min,5min,and 15min(P＜0.01 or P＜0.05),with the peak at the point of 5min(P＜0.01);but concentration of NO did not changed statistically at the point of 30min compared to control group.
②The effects of Apelin on ET-1 release in PMVECs induced by AngiotensinⅡ:
1) Compared with control group,concentrations of ET-1 in cultural medium of AngⅡgroup at three points of 30min,6hr and 24hr increased significantly(P＜0.05).
2) Compared with AngⅡgroup,concentratins of ET-1 in cultural medium of AngⅡ+10~(-8) Apelin group at the three points decreased significantly(P＜0.05),and close to control group.
3) Compared with AngⅡgroup,concentratins of ET-1 in cultural medium of AngⅡ+10~(-6) Apelin group trended to decrease but without statistical significance.
③The effects of Apelin on phosphorylations of endothelial nitric-oxide synthase and Akt:
1) Compared with control group,Apelin induced increase of protein expressions of phosphorylation of both Akt and eNOS significantly in Apelin group(P＜0.01).
2) Compared with Apelin group,protein expressions of phosphorylation of eNOS significantly decreased(P＜0.01).
Conclusion Apelin can induced NO release and attenuated ET-1 release induced by AngⅡin PMEVCs.Promoting phosphorylation of Akt/eNOS was one of mechanisms to induce NO release and may be involved in protective effect on PMVECs for Apelin.
引文
1. Runo JR, et al. Primary pulmonary hypertension. N Engl J Med, 1997,336:111-117.
2. Humbert M, et al. Treatment of Pulmonary Arterial Hypertension. N Engl J Med, 2004,351:1425-1436.
3. Tatemoto, K, et al. Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor. Biochem Biophys Res Commun, 1998,251: 471- 476.
4. Masri B, Morin N, Cornu M, et al. Apelin(65-77) activates p70S6 kinase and is metogenic for umbilical endothelial cells. FASEB J, 2004, 18: 1909-1911.
5. Si YT, Hui X, Ling QY, et al. apelin stimulates proliferation and suppresses apoptosis of mouse osteoblastic cell line MC3T3-E1 via JNK and PI3K/Akt signaling pathways. Peptides, 2007,28: 708-718.
6. Christopher MC, Susan LD, Melanie KM, et al. apelin, the ligand for the endothelial G-protein-coupled receptor, APJ, is a potent angiogenic factor required for normal vascular development of the frog embryo. Developmental Biology, 2006,296: 177-189.
7. Lauren AO, Arpita A, Praveena S, et al. apelin, an endogenous neuronal peptide, protects hippocampal neurons against excitotoxic injury. Journal of Neurochemistry, 2007,102: 1905-1917.
8. Habata, Y, et al. Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrum. Biochim Biophys Acta 1999,1452:25-35.
9. Matthias J. et al. Emerging roles of apelin in biology and medicine. Pharmacology & Therapeutics, 2005,107:198 - 211.
10. Matthias J. et al. Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells. Regulatory Peptides, 2004,118: 119-125.
11.Hosoya M, Kawamata Y, Fukusumi S, et al. Molecular and functional characteristics of APJ. Tissue distribution of mRNA and interaction with the endogenous ligand apelin.J Biol Chem,2000,275:21061-21067.
12.Lee,D.K.,et al.Characterization of apelin,the ligand for the APJ receptor.J Neurochem,2000,74,1:34-41.
13.Tatemoto K,Kiyoshige Takayama,et al.The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism.Regulatory Peptides,2001,99:87-92.
14.Ishida,J,et al.Regulatory roles for APJ,a seven-transmembrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo.J Biol Chem,2004,279:26274-26279.
15.Masri,B,et al.Apelin(65-77) activates p70 S6 kinase and is mitogenic for umbilical endothelial cells.The FASEB,2004,18:1909-1911.
16.Goetze JP,et al.Apelin:A new plasma marker of cardiopulmonary disease.Regulatory Peptides,2006,133:134-138.
17.龚永生等.低氧性肺动脉高压大鼠肺组织apelin-APJ系统的变化。中国微循环,2006,10:119-122.
18.Fulton,D,et al.Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.NATURE,1999,399:597-601.
19.Dimmeler,S,et al.Activation of nitric oxide synthase in endothelial cells byAkt-dependent phosphorylation.NATURE,1999,399:601-605.
1 Runo JR,et al.Primary pulmonary hypertension.N Engl J Med,1997,336:111-117.
2 Gianni A,Losano.On the cardiovascular activity of apelin.Cardiovascular research,2005,65:8-9.
3 Orte C,Polak JM,Haworth SG,et al.Expression of pulmonary vascular argiotension-conventing enzyme in primary and secondary plexiform pulmonary hypertension.J Pathol,2000,192:379-384.
4 Lin LY,Lin CY,Su TC,et al.Angiotensin Ⅱ-induces apoptosis in human endothelial cells is inhibited by adiponectin through restoration of the association between endothelial nitric oxide synthade and heat shock protein 90.FEBS Lett,2004,574:106-110.
5 刘清行,金惠铭.TNFa对大鼠肺微血管内皮细胞NO的产生及NOSmRNA表达的影响.上海医科大学学报,1999,26:4-7.
6 Rohit B,Rubin MT,Paul MH.Endothelial dysfunction in pulmonary hypertension[J].Circulation,2004,109:159-165.
7 Ohuchi H,Takasugi H,Ohashi H,et al.Abnormalities of neurohormonal and cardiac autonomic nervous activities relate poorly to functional status in fontal patients.Circulation,2004,110:2601-2608.
8 Tatemoto K,Hosoya M,Habata Y,et al.Isolatin and characterization of a novel endogenous peptide ligand for the human APJ receptor.Biochem Biophys Res Commun,1998,251:471-476.
9 Klenz MJ,Davenport AP.Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells.Regul Pept,2004,118:119-125.
10 Christopher MC,Susan LD,Melanie M,et al.Apelin,the ligand for the endothelial G-protein-coupled receptor,APJ,is a potent angiogenic factor required for normal vascular development of the frog embryo.Developmental Biology,2006,296:177-189.
11 Atsushi K,Norihito S,Maki O,et al.apelin is a novel angiotenic factor in retinal endothelial cells.Biochemial and Biophysical Research Communication,2004,325:395-400.
12 Cheng X,Cheng XS,Pang CY.Venous dilator effect of apelin,an endogenous peptide ligand for the orphan APJ receptor,in conscious rats.European J Pharmocology,2003,470:171-175.
13 James C,Simpkin SY,Lim SM,et al.apelin,the potentially therapeutic adipocytokine,protects against myocardial ischemia-reperfusion injury.Journal of molecular and cellular cardiology,2007,42:s190-s218.
14 Principe A,Fermandez VG,Melgar LP,et al.apelin,a novel neuroendocrine peptide,plays an important role in the pathogenesis of the cardiovascular dysfunction in cirrhosis.Molecular Biology and Cell Signalling,2007,s51-52.
15 Klara F,Mark AS,Steven RC,et al.Direct effects of apelin on cardiomyocyte contractility and electrophysiology.Biochemical and Bilphysical Research Communications,2007,357:889-895.
16 Jia YX,Pan CS,Zhang J,et al.apelin protects myocardial injury induced by isoproterenol in rats.Regulatory Peptides,2006,133:147-154.
17 Kwok SC,Gardner RS,Morton JJ,et al.Plasma concentrations of the novel peptide apelin are decreased in patients with chronic heart failure.The European Journal of the Heart Failure,2006,8:355-360.
18 Jens PG,Jens FR,Jom C,et al.apelin:a new plasma marker of cardiopulmonary disease.Regulatory Peptides,2006,133:134-138.
19 Bayraktutan U.effects of angiotensin Ⅱ on nitric oxide generation in growing and resting rat aortic endothelial cells.J Hypertens,2003,21:2093-2101.
20 龚永生,范小芳,高钰琪等.缺氧致大鼠心肺组织中apelin的表达降低.第三军医大学学报,2006,28:1188-1190.
21 Ishida 5,Tatsuo H,Yasumi H,et al.Regulatory roles for APJ,a seven-transmenbrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo.rhe Journal of Biological Chemistry,2004,297:26274-26279.
22 Zhong JC,Yu XY,Yu H,et al.apelin modulates aortic vascular tone via endothelial nitric oxide synthase phosphorylation pathway in diabetic mice.Cardiovascular Research,2007,in press.
23 Andy JP,Christine MP.The management of pulmonary hypertension.Pediatric Anesthesia,2006,16:816-821.
1 Klenz MJ,Davenport AP.Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells.Regul Pept,2004,118:119-125.
2 Christopher MC,Susan LD,Melanie KM,et al.Apelin,the ligand for the endothelial G-protein-coupled receptor,APJ,is a potent angiogenic factor required for normal vascular development of the frog embryo.Developmental Biology,2006,296:177-189.
3 Matthias JK,Anthony PD.Emerging roles of apelin in biology and medicine.Pharmacology & Therapeutics,2005,107:198-211.
4 Jens PG,Jens FR,Jorn C,et al.Apelin:A new plasma marker of cardiopulmonary disease.Regulatory Peptides,2006,133:134-138.
5 龚永生,范小芳,高钰琪等.缺氧致大鼠心肺组织中apelin的表达降低.第三军医大学学报,2006,28:1188-1190.
6 Andy JP,Christine MP.The management of pulmonary hypertension.Pediatric Anesthesia,2006,16:816-821.
7 Rohit B,Rubin MT,Paul MH.Endothelial dysfunction in pulmonary hypertension.Circulation,2004,109:159-165.
8 Atsushi K,Norihito S,Maki O,et al.Apelin is a novel angiogenic factor in retinal endothelial cells.Biochemical and Biophysical Research Communications,2004,325:395-400.
9 Masri B,Natacha M,Marion C,et al.Apelin(65-77) activates p70S6kinase and is mitogenic for umbilical endothelial cells.FASEB,2004published online.
10 Ishida J,Tatsuo H,Yasumi H,et al.Regulatory roles for APJ,a seven-transmenbrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo.The Journal of Biological Chemistry,2004,297:26274-26279.
11 金惠铭,卢建,殷莲华主编.细胞分子病理生理学.第1版.郑州:郑州大学出版社,2002,226-230.
12 曾翔俊,王红霞,芦玲巧,等.Apelin-13对离体大鼠心脏缺血/再灌注损伤的影响及机制探讨.中国药理学通报,2007,23:82-5.
13 Euan A,Hyung JC,Thomas Q.Opposing cardiovascular roles for the angiotensin and apelin signaling pathways.Journal of Molecular and Cellular Cardiology,2006,41:778-781.
14 Jeffery TK,Morrell NW.Molecular and cellular basis of vascular remodeling in pulmonary hypertension.Prog Cardiovasc Dis,2002,45:173-202.
1 AndyJP,Christine MP.The management of pulmonary hypertension.Pediatric Anesthesia,2006,16:813-821.
2 Denzil LM,Wilson SC,Michael MG.Secondary pulmonary hypertension in chronic heart failure.Circulation,2000,102:1718-1723.
3 James RR,James EL.Primary pulmonary hypertension.Lancet,2003,361:1533-1544.
4 金惠铭,卢建,殷莲花主编.细胞分子病理生学(M).第1版.郑州大学出版社,2002,226-230.
5 Rohit B,Rubin MT,Paul MH.Endothelial dysfunction in pulmonary hypertension.Circulation,2004,109:159-165.
6 Matthias J,Anthony P.Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells.Regul Pept,2004,118:119-125.
7 Kawamata Y,Habata Y,Fukusumi S,et al.molecular properties of apelin:tissue distribution and receptor binding.Biochim Biophys Acta,2001,1538:162-171.
8 Tatemoto K,Takayama K,Zou MX,et al.The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism.Regul Pept,2001,99:87-92.
9 Cheng X,Cheng XS,Pang CC.Venous dilator effect of apelin,an endogenous peptide ligand for the orphan APJ receptor,in conscious rats.Eur J Pharmacol,2003,470:171-175.
10 Ishida J,Hashimoto T,Hashimoto Y,et al.Regulatory roles for APJ,a seven-transmembrane receptor related to angiotensin-type 1 receptor in blood pressure in vivo.J Biol Chem,2004,279:26274-26279.
11 Llorens S,Nava E.Cardiovascular diseases and the nitric oxide pathway.Curr Vasc Pharmacol,2003,1:335-346.
12 贾月霞,杨靖辉,潘春水,等.Apelin激活大鼠主动脉一氧化氮系统的研究.宁夏医学院报.2005,27:429-431.
13 Gonzalez-Zulueta M,Ensz LM,Mukhina G,et al.Manganese superoxide dismutase protects Nnos neurons from NMDA and nitric oxide-mediated neurotoxicit.J Neurosci,1998,18:2040-2055.
14 莫红缨.NF-κB及iNOS与肺部疾病.国外医学呼吸系统分册,2000,20:160-162.
15 Hawpl V,Jan H..Role of nitric oxide in the pathogenesis of chronic pulmonary hypertension.Physiol Rev,2000,80:1337-1372.
16 Possi GP,Sacchetto A,Cesari M,et al.Interactions between endothelin-1 and the renin-angiotensin-aldosterone system.Cardiovasc Res,1999,43:300-307.
17 Bremnes T,Paasche JD,Mehlum A,et al.Regulation and intracellular traffiching pathways of the endothelin receptors.J Biol Chem,2000,275:17596-17604.
18 Orte C,Polak JM,Haworth SG,et al.Expression of pulmonary vascular argiotension-conventing enzyme in primary and secondary plexiform pulmonary hypertension.J Pathol,2000,192:379-384.
19 俸勇强,伍伟峰.先心病肺动脉高压与血管紧张素Ⅱ的关系研究.右江民族医学院学报,2006,1:10-13.
20 赵静,吴学君,金星,等.血管内皮细胞凋亡诱导剂的研究进展.中国现代普通外科进展,2004,7:70-72.
21 Lin LY,hin CY,Su TC,et al.Angiotensin Ⅱ-induces apoptosis in human endothelial cells is inhibited by adiponectin through restoration of the association between endothelial nitric oxide synthade and heat shock protein 90.FEBS hett,2004,574:106-110.
22 刘玉,孙运峰,马贵喜,等.茶多酚对血管紧张素Ⅱ所致血管内皮细胞分泌内皮素功能的影响.第二军医大学学报,2006,26:1408-1411.
23 Phillips MI,Kagiyama S.Angiotensin Ⅱ as a pro-inflammatory mediator.Curr Opin Investig Drugs,2002,3:569-577.
24 Pueyo ME,Arnal iF,Rami H,et al.Angiotensin Ⅱ stimulates the production of NO and peroxynitric in endothelial cells.Am J Physiol,1998,274:C214-C220.
25 Pueyo ME,Goonazlez W,Nicoletti A,et al.Angiotensin Ⅱ stimulates endothelial vascular cell adhesion molecule-1 via nuclear factor-kappa B activation induced by intracellular oxidative stress.Arterioscler Thromb Vasc Biol,2002,20:645-651.
26 Seshiah PN,Weber DS,Rocic P,et al.Angiotensin Ⅱ stimulation of NADPH oxidase activity:upstream mediators.Circ Res,2002,91:406-413.
27 Christopher MC,Susan LD,Melanie KM,et al.Apelin,the ligand for the endothelial G-protein-coupled receptor,APJ,isa potent angiogenic factor required for normal vascular development of the frog embryo.Developmental Biology,2006,296:177-189.
28 曾翔俊,王红霞,芦玲巧,等.Apelin-13对离体大鼠心脏缺血/再灌注损伤的影响及机制探讨.中国药理学通报.2007,23:82-85.
29 Foldes G,Horkay F,Szokodi I,et al.Circulating and cardiac levels of Apelin,the novel ligand of the orphan receptor APJ,in patients with heart failure.Biochem Biophys Res Commun,2003,308:480-485.
30 金戈,杨鹏麟,龚永生,等.两肾一夹高血压大鼠左心室apelin及其受体的基因表达.温州医学院学报,2007,37:206-209.
31 龚永生,范小芳,高玉琪,等.缺氧致大鼠心肺组织中apelin的表达降低.第三军医大学学报,2006,28:1188-1190.
32 Chen MM,Ashley EA,Deng DX,et al.Novel role for the potent endogenous inotrope apelin in human cardiac dysfunction.Circulation,2003,108:1432-1439.
33 Goetze JP,Rehfeld JF,Carlsen J,et al.apelin:A new plasma marker of cardiopulmonary disease.Regul Pept,2006,133:134-138.
34 Francia P,Salvati A,Balla C,et al.Cardiac resynchronization therapy increases plasma levels of the endogenous inotrope apelin.European Journal of Heart Failure,2007,9:306-309.
35 和亚萍,魏盟,刘铭雅,等.大鼠心肌梗死后早期Apelin的表达上调.中国临床医学,2007,4:462-464.
36 Fulton,D,et al.Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.NATURE,1999,399:597-601.
37 Dimmeler,S,et al.Activation of nitric oxide synthase in endothelial cells byAkt-dependent phosphorylation.NATURE,1999,399:601-605.
38 Masri B,Natacha M,Marion C,et al.Apelin(65-77) activates p70S6kinase and is mitogenic for umbilical endothelial cells.FASEB,2004published online.
39 Zhu M,Feng JH,Eliana L,et al.Ischemic postconditioning protects remodeled myocardium via the PI3K-PKB/Akt reperfusion injury salvage kinase pathway.Cardiovascular Research,2006,72:152- 162.
40 Si YT,Hui X,Ling QY,et al.apelin stimulates proliferation and suppresses apoptosis of mouse osteoblastic cell line MC3T3-E1 via JNK and PI3K/Akt signaling pathways.Peptides,2007,28:708-718.
41 Lauren AO,Arpita A,Praveena S,et al.apelin,an endogenous neuronal peptide,protects hippocampal neurons against excitotoxic injury.Journal of Neurochemistry,2007,102:1905-1917.
42 余洋,黄德嘉.Apelin对心肌细胞肥大的影响.心脏杂志,2006,18:185-189.
43 Jia YX,Pan CS,Zhang J,et al.apelin protects myocardial injury induced by isoproterenol in rats.Regul Pept,2006,133:147-54.
44 James CS,Shiang YL,Sean MD,et al.apelin,the potentially therapeutic adipocytokine,protects against myocardial ischemia-reperfusion injury.Journal of Molecular and Cellular Cardiology,2007,3:630.
45 Matthias J,Kleinz 6F,Baxter.apelin reduces myocardial reperfusion injury independent of PI3K/Akt and P70S6K.Journal of Molecular and cellular Cardiology,2007,42:s190-s218.
1 Wilson S, Bergsma DJ, Chambers JK, et al. Orphan G-protein-coupled receptors: the next generation of drug targets. Br J Pharmacol, 1998,125:1387-1392.
2 Howard AD, McAllisterG, Feighner SD, et al. Orphan G-protein-coupled receptors and natural ligand discovery. Trends Pharmacol Sci, 2001, 22 : 132-140.
3 Civelli O, Nothacker HP, Saito Y, et al. Novel neurotransmitters as natural ligands of ophan G-protein-mcoupled receptors. Trends Neurosci,2001, 24: 230-237.
4 O'Dowda BF, Heiberb M, Chan A, et al. A human gene that shows identity with the gene encoding the angiotensin receptor is located on chromosome 11. Gene, 1993, 136: 355-360.
5 Tatemoto K, Hosoya M, Habata Y, et al. Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor. Biochem Biophy Res commun, 1998, 251: 471-476.
6 Lee DK, Cheng R, Nguyen T, et al. characterization of Apelin, the ligand for the APJ receptor. J Neurochem, 2000, 74: 34-41.
7 Habata Y, Fujii R, Hosoya M, et al. Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrums. Biochim Biophys Acta, 1999, 1452: 25-35.
8 Medhurst AD, Jennings CA, Robbins MJ, et al. Pharmacological and immunohistochemical characterization of the APJ receptor and its endogenous ligand Apelin. J Neurochem, 2003, 84: 1162-1172.
9 De Mota N, Lenkei Z, LlorensCortes C. Cloning, pharmacological characterization and brain distribution of the rat Apelin receptor. Neuroendocrinology,2000,72: 400-407.
10 Kazuhiko T, Masaki H, Yugo H, et al. Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor. Biochemical and Biophysical Research Communications,1998, 251: 471-476.
11 Christopher MC, Susan LD, Melanie KM, et al. Apelin, the ligand for the endothelial G-protein-coupled receptor, APJ, is a potent angiogenic factor required for normal vascular development of the frog embryo. Development Biology,2006, 296: 177-189.
12 Matthias JK, Anthony PD. Emerging roles of apelin in biology and medicine. Pharmacology & Therapeutics, 2005, 107: 198-211.
13 Ashley E, Chun H, Thomas Q. Opposing cardiovascular roles for the angiotensin and apelin signaling pathways. Journal of Molecular and Cellular Cardiology, 2006,41: 778-781.
14 Lee DK, Cheng R, Nguyen T, et al. Characterization of apelin, the ligand for the APJ receptor. Journal of Neurochemistry, 2000, 74: 34-41.
15 Oudit GY, Crackower MA, Backx PH, et al. The role of ACE2 in cardiovascular physiology. Trends Cardiovasc Med, 2003, 13: 93-101.
16 Vickers C, Hales P, Kaushik V, et al. Hydrolysis of Biological Peptides by Human Angiotensin-converting Enzyme-related carboxypeptidase. J Biol Chem, 2002, 277: 14838-14843.
17 O' Dowda BF, HeigerbM, Chan, A, et al. A human gene that shows identity with the gene encoding the andiotensin receptor is located on chromosome 11. Gene, 1993, 136: 355-360.
18 Devic E, Rizzoti K, Bodin S, et al. Amino acid sequence and embryonic expression of msr/apj, the mouse homolog of Xenopus Xmsr and human APJ. Mech Dev, 1999, 84: 199-203.
19 Hosoya M, Kawamata Y, Fukusumi S, et al. Molecular and functional characteristics of APJ, Tissue distribution of mRNA and interaction with the endogenous ligand Apelin. J Biol Chem, 2000, 275:21061-21067.
20 Zhou NM, Zhang XL, Fan XJ, et al. The N-terminal domain of APJ, a CNS-based coreceptor for HIV-1, is essential for its receptor function and coreceptor activity. Virology, 2003, 317: 84-94.
21 Wang G, Anini Y, Wei W, et al. Apelin, a new enteric peptide: locatization in the gastrointestinal tract, ontogeny, and stimulation of gastric cell proliferation and of cholecystokinin secretrion. Endocrinology, 2004,145: 1342-1348.
22 Habata Y, Fujii R, Hosoya M, et al. Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrums. Biochim Biophys Acta, 1999, 1452: 25-35.
23 Kawamata Y, Habata Y, Fukusumi S, et al. molecular properties of apelin: tissue distribution and receptor binding. Biochim Biophys Acta, 2001, 1538: 162-171.
24 O' Carroll AM, Selby TL, Palkovits M, et al. Distribution of mRNA encoding B78/apj, the rat homologue of the human APJ receptor, and its endogenous ligand apelin in brain and peripheral tissue. Biochim Biophys Acta, 2000, 1492:72-80.
25 Tatemoto K, Takayama K, Zou MX, et al. The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism. Regul Pept, 2001,99: 87-92.
26 Susaki E, Wang G, Cao G, et al. Apelin cells in the rat stomach. Regul Pept, 2005, 129: 37-41.
27 Reaux A, Gallatz K, Palkovits M, et al. Distribution of apelin-synthesizing neurons in the adult rat brain. Neuroscience. 2002,113: 653-662.
28 Reaux A, De MN, Skultetyova I, et al. Physiological role of a novel neuropeptide, apelin , and its receptor in the rat brain. J Neurochem, 2001,77: 1085-1096.
29 Brailoiu GC, Dun SL, Yang J, et al. Apelin-immunoreactivity in the rat hypothalamus and pituitary. Neurosci Lett, 2002, 327: 193-197.
30 Kleinz MJ, Davenport AP. Immunocytochemical localization of the endogenous vasoactive peptide apelin to human vascular and endocardial endothelial cells. Rugul Pept, 2004, 118: 119-125.
31 Foldes G, Horkay F, Szokodi I, et al. Circulating and cardiac levels of apelin, the novel ligand of the orphan receptor APJ, in patients with heart failure. Biochem Biophys Res Commun, 2003, 308:480-485.
32 Kleinz MJ, Skepper JN, Davenport AP. Immunocytochemical localization of the apelin receptor, APJ, to human cardiomyocytes, vascular smooth muscle and endothelial cells. Regul Pept, 2005, 126: 233-240.
33 Matsumoto M, Hidaka K, Akiho H, et al. Low stringency hybridization study of the dopamine D4 receptor revealed D4-like mRNA distribution of the orphan seventransmembrane receptor, APJ, in human brain. Neurosci Lett, 1996, 219(:119-122.
34 Sorhede WM, Magnusson C, Ahren B. The APJ receptor is expressed in pancreatic islets and its ligand, Apelin, inhibits insulin secretion in mice. Regul Pept, 2005, 131: 12-17.
35 Edinger AL, Hoffman TL, Sharron M, et al. An orphan seven-transmembrane domain receptor expressed widely in the brain functions as a coreceptor for human immunodeficiency virus type 1 and simian immunodeficiency virus. J Virol, 1998, 72: 7934-7940.
36 Puffer BA, Sharron M, Coughlan CM, et al. Expression and coreceptor function of APJ for primate immunodeficiency viruses. Virology, 2000,276: 435-444.
37 Choe W, Albright A, Sulcove J, et al. Functional expression of the seven-transmenbrane HIV-1 coreceptor APJ in neural cells. J Neurovirol, 2000, 6, suppl 1: s61-69.
38 Chen MM, Ashley EA, Deng DX, et al. Novel role for the potent endogenous inotrope apelin in human cardiac dysfunction. Circulation, 2003, 108: 1432-1439.
39 Zhou N, Fan X, Mukhtar M, et al. Cell-cell fusion and internalization of the CNS-based, HIV-1 coreceptor, APJ. Virology,2003, 307: 22-36.
40 Ghosh RN, Maxfield FR. Evidence for nonvectorial, retrograde transferring trafficking in the early endosomes of HEp2 cells. J Cell Biol, 1995, 128: 549-561.
41 Pizard A, Blaukat A, Muller EW, et al. Bradykinin-induced internalization of the human B2 receptor requires phosphorylation of three serine and two threonine residues at its carboxyltail. J Biol Chem, 1999, 274: 12738-12747.
42 Christopher MC, Susan LD, Melanie KM, et al. apelin, the ligand for the endothelial G-protein-coupled receptor, APJ, is a potent angiogenic factor required for normal vascular development of the frog embryo. Developmental Biology, 2006,296: 177-189.
43 Si YT, Hui X, Ling QY, et al. apelin stimulates proliferation and suppresses apoptosis of mouse osteoblastic cell line MC3T3-E1 via JNK and PI3K/Akt signaling pathways. Peptides, 2007,28: 708-718.
44 Lauren AO, Arpita A, Praveena S, et al. apelin, an endogenous neuronal peptide, protects hippocampal neurons against excitotoxic injury. Journal of Neurochemistry, 2007,102: 1905-1917.
45 Kasai A,Shintani N,Dda M,et al.Apelin is a novel angiogenic factor in retinal endothelial cell.Biochem Biophys Res Commun,2004,325:395-400.
46 Masri B,Morin N,Cornu M,et al.Apelin(65-77) activates p70S6 kinase and is metogenic for umbilical endothelial cells.FASEB J,2004,18:1909-1911.
47 李锋,李兰芳,秦旭平,等.Apelin-13促进血管平滑肌细胞增殖作用研究.中国药理学通报,2007,23:949-53。
48 Ishida J,Hashimoto T,Hashimoto Y,et al.Regulatory roles for APJ,a seven-transmembrane receptor related to angiogensin-type 1 receptor in blood pressure in vivo.J Biol Chem,2004,279:26274-26279.
49 Cheng X,Cheng XS,Pang CC.Venous dilator effect of apelin,an endogenous peptide ligand for the orphan APJ receptor,in conscious rats.Eur J Pharmacol,2003,470:171-175.
50 Katugampola SD,Maguire JJ,Matthewson SR,et al.[(125)I]-(Pyr1)Apelin-13 is a novel radioligand for locatizing the APJ orphan receptor in human and rat tissues with evidence for a vasoconstrictor role in man.Br J Pharmacol,2001,132:1255-1260.
51 Zhong JC,Huang DY,Liu GF,et al.Effects of all-trans retinoic acid on orphan receptor APJ signaling in spontaneously hypertensive rats.Cardiovasc Res,2005,65:743-50.
52 Adely S,Jesus G,Luis M,et al.apelin effects in human splanchnic arteries.Role of nitric oxide and prostanoids.Regulatory Peptides,2007,708:1-6.
53 Zhong JC,Yu H,Yung LM,et al.The novel peptide apelin regulated intrarenal artery tone in diabetic mice.Regulatory Peptides,2007,06:010.
54 Zhong JC,Yu H,Yung LM,et al.hpelin modulates aortic vascular tone via endothelial nitric oxide synthase phosphorylation pathway in diabetic mice.Cardiovascular Research,2007,doi:10.1016.
55 Kagiyama S,Fukuhara M,Matsumura K,et al.Central and peripheral cardiovascular actions of apelin in conscious rats.Regul Pept,2005,125:55-59.
56 De Mota N,Reaux-Le GA,Messari S,et al.Apelin,a potent diuretic neuropeptide counteracting vasopressin actions through inhibition of vasopressin neuron activity and vasopressin release.Proc Natl Acad Sci USA,2004,101:10464-10469.
57 Seyedabadi M,Goodchild AK,Pilowsky PM.Site-specific effects of apelin-13 in the rat medulla oblongata on arterial pressure and respiration.Auton Neurosci,2002,101:32-38.
58 Charles C,J Rademaker Mr,Richards AM.Apelin-13 induces a biphasid haemodynamic respionse and hormonal activation in normal conscious sheep.J Endocrinol,2006,189:701-10.
59 Szolodi I,Tavi P,Foldes G,et al.Apelin,the novel endogenous ligand of the orphan receptor APJ,regulates cardiac contractility.Circ Res,2002,91:434-440.
60 Berry MF,Pirolli rJ,Jayasankar V,et al.Apelin has in vivo inotropic effects on normal and failing heart.Circulation,2004,110:187-93.
61 Farkasfalvi K,Stagg MA,Coppen SR,et al.Direct effects of apelin on cardiomyocyte contractility and electrophysiology.Biochem Biophys Res Commun,2007,357:889-95.
62 Tieying D,Genaro RC,Wei DG.Apelin increases contractility in failing cardiac muscle.European Journal of Pharmacology,2006,553:222-228.
63 Ashley EA,Powers J,Chen M,et al.The endogenous peptide apelin potently improves cardiac contractility and reduces cardiac loading in vivo.Cardiocasc Res,2005,65:73-82.
64 Jia YX,Pan CS,Zhang J,et al.apelin protects myocardial injury induced by isoproterenol in rats.Regul Pept,2006,133:147-54.
65 张明,同静,应斌宇,等.大鼠心肌缺血/再灌注损伤及预处理apelin的表达.中国循环杂志,2007,11:296-300.
66 曾翔俊,王红霞,芦玲巧,等.Apelin-13对离体大鼠心脏缺血/再灌注损伤的影响及机制研究.中国药理学通报,2007,23:82-5.
67 余洋,黄德嘉.Apelin对心肌细胞肥大的影响.心脏杂志,2006,18:185-189.
68 James CS,Shiang YL,Sean MD,et al.apelin,the potentially therapeutic adipocytokine,protects against myocardial ischemia-reperfusion injury.Journal of Molecular and Cellular Cardiology,2007,3:630.
69 Matthias J,Kleinz GF,Baxter.apelin reduces myocardial reperfusion injury independent of PI3K/Akt and PTOS6K.Journal of Molecular and cellular Cardiology,2007,42:s190-s218.
70 Saint-Geniez M,Masri B,Malecaze F,et al.Expression of the murine msr/apj receptor and its ligand apelin is up regulated during formation of the retinal vessels.Mech Dev,2002,110:183-186.
71 Sorli SC,Le GC,Knibiehler B,et al.apelin is a potent activator of tumour neoangiogenesis.Oncogene,2007,26:7692-7699.
72 贾月霞,杨靖辉,潘春水,等.Apelin激活大鼠主动脉一氧化氮系统的研究.宁夏医学院学报,2005,27:429-431.
73 贾月霞,杨靖辉,潘春水,等.Apelin激活大鼠主动脉阳离子氨基酸转运体的表达.宁夏医学院学报.2005,27:655-657.
74 David F,Jean PG,Timothy JM,et al.Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.Nature,1999,399:597-602.
75 Strfenie D,Ingrid F,Beate F,et al.Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation.Nature,1999,399:601-606.
76 顾虹,王红霞.Apelin心血管作用及其信号转导途径.世界急危重病医学杂志,2006,3:1414-1416.
77 Masri B,Lahlou H,Mazarguil H,et al.apelin(65-77)activates extracellular signal-regulated kinases via a PTK-sensitive G protein.Biochem Biophys Res Commun,2002,209:539-545.
78 Nisoli E,Clemeti E,Paduoci C,et al.Mitochondrial biogenesis in mamals the role of endogenous nitric oxide.Science,2003,299:896-899.
79 Hashimoto T,Kihara M,Ishida J,et al.Apelin stimulates myosin light chain phosphorylation in vascular smooth muscle cells.Arterioscler Thromb Vasc Biol,2006,26:1267-1272.
80 Taheri S,Murphy K,Cohen M,et al.The effects of centrally administered apelin-13 on food intake,water intake and pituitary hormone release in rats.Bilchem Biophys Res Commun,2002,291:1208-1212.
81 Jaszberenyi M,Bujdoso E,Telegdy G.Behavioral,neuroendocrine and thermoregulatory actions of apelin-13.Neuroscience,2004,129: 811-816.
82 O' Carroll AM, Lolait SJ. Regulation of rat APJ receptor messenger ribonucleic acid ecpression in magnocellular neurons of the paraventricular and suspraoptic nuclei by osmotic stimuli. J Neuroendocrinol, 2003,15: 661-666.
83 O' Carroll AM, Don AL, Lolait SJ. APJ receptor mRNA expression in the rat hypothalamic paraventricular nucleus: regulation by stress and glucocorticoids. J Neuroendocrinol, 2003, 15: 1095-1101.
84 O' Shea M, Hansen MJ, Tatemoto K, et al. inhibitory effect of apelin-12 on nocturnal food intake in the rat. Nutr Neurosci, 2003,6:163-167.
85 Sunter D, Hewson AK, Dickson SL, et al. Intracerebrventricular injection of apelin-13 reduces food intake in the rat. Neurosci Lett, 2003, 353:1-4.
86 Valle A, Hoggard N, Adams AC, et al. chronic central administration of apelin-13 over 10 days increases food intake, body weight, locomotor activity and body temperature in C57BL/6 Mice. Journal of Neuroendocrinology, 2007,20: 79-84.
87 Boucher J, Masri B, Daviaud D, et al. Apelin, a newly identified adipokine up-regulated by insulin and obesity. Endocrinology, 2005,146: 1764-1771.
88 Wei L, Hou X, Tatemoto K. Regulation of apelin mRNA expression by insulin and glucocorticoids in mouse 3T3-L1 adipocytes. Regul Pept, 2005, 132: 27-32.
89 Daviaud D, Boucher J, Gesta S, et al. TNF alpha up-regulates apelin expression in human and mouse adipose tissue. FASEB J, 2006,20: 1528-1530.
90 Horiuchi Y, Fujii T, Kamomura Y, et al. The endogenous, immunologically active peptide apelin inhibits lymphocytic cholinergic activity during immunological responses. Neuroimmunol, 2003, 144: 46-52.
91 Hus-Citharel A, Bouby N, Ituir joz X, et al. Distribution of apj receptor mRNA expression along the ramephron evidence for a vasoconstrictor role of apelin in glomerular arerioles. Hypertension, 2005,46:905.
92 Katugampola S, Davenport A. Emerging roles fro orphan G protein-coupled receptors in the cardiovascular system.Trends Pharmacol Sci,2003,24:30-35.
93 金戈,杨鹏麟,龚永生,等.两肾一夹高血压大鼠左心室apelin及其受体的基因表达.温州医学院学报,2007,37:206-209.
94 Ellinor PT,Low AF,Macrae CA.Reduced apelin levels in lone atrial fibrillation.Eur Heart J,2006,27:222-226.
95 张明,童静,应斌宇,等.冠心病患者血浆apelin水平的变化.实用医学杂志,2007,23:3361-3362。
96 Keiji K,Liyong Z,Yumiko L,et al.impaired heart contractility in apelin gene-deficient mice associated with aging and pressure overload.Circ Res,2007,101:e32-e42.
97 Chong KS,Gardner RS,Morton JJ,et al.Plasma concentrations of the novel peptide apelin are decreased in patients with chronic heart failure.Eur J Heart Fail,2006,Feb3:[Epub ahead of print].
98 Malyszko J,Malyszko JS,Kozminski P,et al.Apelin and cardiac function in hemodialyzed patients:possible relations.Am J Nephrol,2006,26:121-126.
99 Goetze JP,Rehfeld JF,Carlsen J,et al.apelin:A new plasma marker of cardiopulmonary disease.Regul Pept,2006,133:134-138.
100 Francia P,Salvati A,Balla C,et al.Cardiac resynchronization therapy increases plasma levels of the endogenous inotrope apelin.European Journal of Heart Failure,2007,9:306-309.
101 龚永生,范小芳,吴小脉等.低氧性肺动脉高压大鼠肺组织apelin-APJ系统的变化.中国微循环,2006,10:119-123.
102 项中一,李玉萍,陈少贤等。肺动脉高压患者血浆apelin的变化及意义.放射免疫学杂志,2006,19:262-264.
103 Heinonen MV,Purhonen AK,Miettinen P,et al.Apelin,orexin-A and leptin plasma levels in morbid obesity and effect of gastric banding.Regul Pept,2005,130:7-13.
104 Castan-Laurell I,Boucher J,Dray C,et al.Apelin,a novel adipokine over-produced in obesity:friend or foe? Mol Cell Endocrinol,2005,245:7-9.
105 Weisberg SP,McCann D,Desai M,et al.Obesity is associated with macrophage accumulation in adipose tissue.J Clin Invest,2003,112: 1796-1808.
106 Hotamisligil GS,Murray DL,Choy LN,et al.Tumor necrosis factor alpha inhibits signaling from the insulin receptor.Pro Natl Acad Sci USA,1994,91:4854-4858.
i07 Farzan M,Choe H,Martin K,et al.Two orphan seven-transmembrane segment receptors which are expressed in CD4-positive cells support simian immunodeficiency virus infection.J Exp Med,1997,186:405-411.
108 Choe H,Farzan M,Sun Y,et al.The beta-chemokine receptors CCR3 and CCR5 facilitate infection by primary HIV-1 isolates.Cell,1996,85:1135-1148.
109 Dragic T,Litwin V,Allaway GP,et al.HIV-1 entry into CD4+ cells is mediated by the chemokine receptor CC-CKR5.Nature,1996,381:667-673.
110 Choe H,Farzan M,Konkel M,et al.The orphan seven-transmenbrane receptor apj supports the entry of primary T-cell-line tropic and dualtropic human immunodeficiency virus type 1.J Virol,1998,72:6113-6118.
111 Esposito V,De Falco M,De Luca L,et al.Functional expression of the seventransmembrane HIV-1 co-receptor APJ in neural cells.J Neurovirol,2000,6:S61-S69.
112 Zou MX,Liu HY,Haraguchi Y,et al.Apelin peptides block the entry of human immunodeficiency virus(HIV).FEBS Lett,2000,473:15-18.
113 Cayabyab M,Hinuma S,Farzan M,et al.Apelin,the natural ligand of the orphan seven-transmmbrane receptor APJ,inhibits human immunodeficiency virus type 1 entry.J Virol,2000,74:11972-11976.