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血管紧张素 和血小板源生长因子对心肌细胞周期素和p27蛋白表达量的调节 |
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袁 勇 许顶立 刘伊丽 贾满盈
摘 要 目的：细胞周期素(cyclin)和CDK抑制蛋白(CKI)是对细胞周期起正性和负性调节作用的两类物质。p27蛋白是CKI的一种，主要对细胞外部促进或抑制增殖的信号起反应。本文观察血管紧张素Ⅱ(AngⅡ)和血小板源生长因子(PDGF)对心肌细胞细胞周期素和p27蛋白表达量的影响。方法：培养的心肌细胞除血清24 h使其处于静止状态，加入AngⅡ10-6mol/L、PDGF-BB20 ng/mL后不同时间收集细胞，用碘化丙啶(PI)标记细胞DNA，以确定细胞所处的周期。用细胞周期素或p27蛋白的单抗和标记FITC的二抗标记细胞，用流式细胞仪测定被激发出的荧光量来测定细胞周期素和p27蛋白表达的相对量。结果：AngⅡ可促进细胞周期素的表达，但对p27蛋白的表达没有影响，不能促进心肌细胞增殖；PDGF明显促进心肌细胞细胞周期素的表达，也不能明显抑制p27的表达，也不能促进心肌细胞增殖。结论：细胞周期素的表达增加与心肌细胞能否进入细胞周期并不一致，p27蛋白是心肌细胞能否进入细胞周期并进行有丝分裂的重要调节因子。
主题词 细胞周期蛋白类；心肌；细胞周期；流式细胞术
Angiotensin Ⅱ and platelet derived growth factor regulate the expression of cyclins and cyclin dependent kinase inhibitor p27 in cardiac myocytes
YUAN Yong, XU Ding-Li, LIU Yi-Li, JIA Man-Ying
Department of Cardiology, Nanfang Hospital, The First Military Medical University, Guangzhou (510515)
Abstract AIM: To investigate cell cycle distribution of cardiac myocytes(CM) and expression of cyclins and p27 (one of cyclin dependent kinase inhibitors) caused by angiotensin Ⅱ(AngⅡ) and platelet derived growth factor BB(PDGF-BB). METHODS: Cultured CMs were deprived of fetal calf serum for 24 hours to make quiescent. CMs were collected at different times after stimulation of AngⅡand PDGF-BB. Cell cycle distribution and expression of cyclins and p27 were determined with flow cytometer. RESULTS: AngⅡincreased expression of cyclins, but had no effects on expression of p27. PDGF could enhanced expression of cyclins significantly, but did not inhibit p27 expression. Both could not promote CM proliferation. CONCLUSION: Enhanced expression of cyclins was not correlated to progression of CM cell cycle. p27 inhibits CM progression through G1 to S phase and thus inhibited CM proliferation.
MeSH Cyclins; Myocardium; Cell cycle; Flow cytometer
心肌细胞(cardiac myocyte,CM)数量的减少是心力衰竭产生和/或恶化的最主要的病理基础［1］。一般认为成年的哺乳动物心肌细胞属于终末分化细胞，很少或几乎不能进行有丝分裂，然而最近发现在成年急性和慢性心力衰竭动物模型以及心力衰竭患者心脏都观察到心肌细胞增殖的现象［2］，促进[1] [2] 下一页
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