摘要
目的:观察血管紧张素转换酶抑制剂(ACEI)依那普利拉(Ena)和蛋白激酶C(PKC)抑制剂白屈菜红(Chele)对血管紧张素Ⅱ(AngⅡ)诱导的新生鼠心肌成纤维细胞(CFb)增殖、细胞周期、Ⅰ型胶原纤维(collagenⅠ)、PKC和细胞周期蛋白cyclinD1蛋白表达的影响,阐明Ena抗CFb增殖的分子机制。方法:培养的新生Wistar大鼠CFb分为对照组、AngⅡ组、Chele+AngⅡ组、Chele+AngⅡ+Ena组和AngⅡ+Ena组,采用胰酶消化、差速贴壁法培养CFb,四氮唑盐(MTT)比色法检测细胞增殖;免疫细胞化学染色(IC)法测定collagenⅠ含量;流式细胞术(FCM)检测细胞周期;免疫印迹法检测PKC和细胞周期蛋白cyclinD1表达。结果:与AngⅡ组比较,Chele和Ena组及Chele+AngⅡ+Ena组CFb增殖率均显著降低(P<0.05或P<0.001),collagenⅠ含量降低(P<0.05或P<0.01),CFb G0/G1期细胞百分率升高,S期细胞百分率降低(P<0.05或P<0.01),PKC和cyclinD1蛋白表达水平降低(P<0.05或P<0.01)。结论:Ena和Chele能抑制AngⅡ诱导的CFb增殖和胶原蛋白分泌,其机制可能是通过抑制PKC-cyclinD1转导通路实现的。
Objective To observe the effects of ACE inhibitor enalaprilat（Ena） and protein kinase C（PKC） inhibitor chelerythrine（Chele）on proliferation,collagen Ⅰ,cell cycle,PKC and cyclinD1 protein expression of neonatal cardiac fibroblasts（CFb） and to probe its molecular mechanism.Methods CFb of neonatal Wistar rats were divided into control group,AngⅡ group,Chele＋AngⅡ group,Chele＋AngⅡ＋Ena group and AngⅡ＋Ena group.CFb were isolated by trypsin digestion method.MTT colorimetric assay was adopted to evaluate cell proliferation,immunocytochemical staining（IC） was used to measure collagen Ⅰ content,Western blotting and flow cytometry were used to detect PKC,cyclinD1 and cell cycle respectively.Results Compared with AngⅡ group,the MTT value decreased dramatically（P〈0.05 or P〈0.001）,collagen Ⅰ content decreased（P〈0.05 or P〈0.01）;the percentage of cell at G0/G1 phase increased and the percentage of cells at S phase decreased（P〈0.05 or P〈0.01）;and the PKC and cyclinD1 protein expressions were inhibited in Chele,Ena,and Chele＋AngⅡ＋Ena groups（P〈0.05 or P〈0.01）.Conclusion The antiproliferation effects of Ena on CFb can be attributed to inhibiting PKC/cyclinD1 signal pathway possibly.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2009年第4期591-594,F0002,共5页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(30672654)
作者简介
孙红霞（1965-），女，吉林省吉林市人，教授，医学博士，主要从事心血管分子药理学的研究。
[通讯作者]杨世杰（Tel：0431—85619483，E-mail：jcyaoli@sina．com）
