et1调控缺氧诱导anp分泌的磷脂酰肌醇3酶信号转导机制研究.docxVIP

优秀毕业论文 精品参考文献资料 延垫太堂亟± 延垫太堂亟±堂僮途塞 擅 噩 摘 要 目的：缺氧明显促进心房内皮素．1(endothelin．1，ET-1)的生成及其释放，并参 与调节心房功能。然而，内源性ET-1NJ控心房钠尿肽(atrial natriuretic peptide． ANP)分泌的具体机制尚不甚清楚。因此，本研究利用分子生物学、放射免疫、 药理学等技术观察和探讨内源性ET-1调控缺氧诱导ANP分泌的磷脂酰肌醇．3． 激酶(phosphatidylinositol一3一kinase，P13K)／蛋白激酶B(protein kinase B，Akt) 信号转导机制。 方法：本研究利用离体搏动的家兔心房灌流装置制备急性缺氧模型，采用放射免 疫技术检测ET-1及ANP的含量，选用蛋白印迹技术(westernblot，WB)分析内 皮素A型和B型受体(endothelin receptor type A and B，ETRA and ETRB)的表达 和Akt及GATA结合蛋白．4(GATAbiding protein．4，GATA一4)含量的变化。 结果：1．急性缺氧明显促进家兔心房ET-1的释放(n=6，P0．001 VS．control period)，并显著上调ETRA及ETRB表达(n=5，P0，00 1 and P0．0 1 VS．control group)。同时显著增加心房ANP的分泌，并明显抑制心房搏出量和搏动压(n=6， P0．001 VS．control period)。2．ETRA和ETRB的抑制剂BQl23(O．3 pmol／L)及 BQ788(0．3 gmol／L)明显减缓缺氧诱导ANP分泌的效应(11=6，PO．01 VS．control period)。3．P13K／Akt的抑制剂LY294002(30．0 gm01)也可明显抑制缺氧诱导 ANP分泌的作用(n=6，P0．001 VS．hypoxia)。4．急性缺氧明显增加磷酸化Akt (phospho Akt，pAkt)和GATA4的含量(均n=5，P0．001 VS．contr01)。5．BQl23 和BQ788则可明显减缓缺氧增加pAkt(分别n=5，P0．05；P0．01 VS．hypoxia) 及GATA4(分别n=5，P0．05；P0．001 VS．hypoxia)含量的效应。 结论：内源性ET-1通过ETR．P13K／Akt．GATA4信号转导通路调节缺氧诱导家兔 心房ANP分泌的过程。 关键词：缺氧：内皮素．1；心房钠尿肽；信号通路；GATA4结合蛋白 万方数据 延垫太堂亟± 延垫太堂亟±堂焦途塞一．． △坠§!堕丛 Abstract Aim：Hypoxia promotes atrial ET-1 production and contributes to regulation of hypoxia-induced atrial natriuretic peptide(ANP)secretion．However,the mechanism by which hypoxia-induced endogenous ET-1 regulates ANP secretion is remains to be defme．The purpose of the present study was to investigate the mechanism that the endogenous ET-1 regulates atrial ANP secretion． Methods：Isolated perfused acute hypoxic rabbit atrial model was used in this study． The levels of ET-1 and ANP in the atrial perfusates were measured by radioimmunoassay，and the protein levels of ET receptors，phosphorylated protein kinase B(pAU、and GATA-4 protein in atrial tissue were determined by Western blot． Results：1．Acute hypoxia significantly enhanced ET-1 release and up regulated bo