磷酸二酯酶抑制剂对乳鼠心肌细胞PI3K—Akt—eNOS信号通路的影响.pdfVIP

内科急危重症杂志 2008年 第14卷 第3期 139 磷酸二酯酶抑制剂对乳鼠心肌细胞PI3 K—Akt—eNOS 信号通路的影响 华中科技大学同济医学院附属同济医院 周名纲 马业新 张新金，武汉 430030 摘要 目的：探讨西洛他唑(Cilostazo1)对乳鼠心肌细胞PI3K—Akt-eNOS信号通路的影响。方法：观察西洛他 唑对乳鼠心肌细胞NO影响的时效和量效关系，再用PI3K及 eNOS抑制剂进行干预。检测NO的浓度，Western 免疫印迹法检测总Akt、磷酸化 Akt(p—Akt—Ser473)及总eNOS、磷酸化eNOS(p-eNOS~SerlI77)表达水平。结果： 西洛他唑升高心肌细胞NO浓度呈剂量和时间依赖性，不同浓度的西洛他唑均能升高Akt和eNOS磷酸化水平， 但对Akt及eNOS总蛋白表达无明显影响。eNOS抑制剂L—NAME和PI3K抑制剂Wortmannin均能抑制西洛他 唑诱导的NO浓度升高，Wortmannin还能阻断西洛他唑诱导的Akt和eNOS的磷酸化。结论：西洛他唑可能激活 乳鼠心肌细胞的PI3K—Akt—eNOS信号通路而促进 N0的产生。 关键词 西洛他唑 心肌细胞 磷脂酰肌醇一3激酶 丝氨酸／苏氨酸蛋白激酶 一氧化氮 内皮型一氧化氮 合酶 中图分类号 R54I 文献标识码 A Effect of Phosphodiesterase Inhibitor on PI3K-Akt-eNOS Pathway in Cardiomyocytes of Neonatal Rat ZHOU Ming— gang，MA Yexin，ZHANG Xinjin．Tongji Hospital Affiliated to Tongji Medical College，Huazhong University of Science and Technology，Wuhan 430030，China Abstract Objective：To investigate the effect of Cilostazol on phosphatidy1inosito1—3 kinase(PI3K)／Serine／ threonine protein kinase(AKT)／endothelial nitric oxide synthase(eNOS)pathway in cardiomyocyte of neonatal SD rat．Methods：To observe the time-effect and dose-effect relationship of Cilostazo1 on nitric oxide(NO)concentration in cardiomyocytes of cultured neonatal rat．Expressions of Akt，phospho—Akt and eNOS，phospho—eNOS were de— tected by Western blot．Results：Cilostazo1 increased the endothelia1 NO production in a dose—and time—dependent manner in cardiomyocytes of cultured neonata1 SD rat and also increased the expression of phosphory1ation of Akt and eNOS in a concentration—dependent manner，with no alteration in the expression of total Akt and total eNOS in cultured cardiomyocytes． L—NAME，eNOS synthase inhibitor，and Wortmannin，PDK inhibitor，blocked the