苦参碱对醛固酮诱导大鼠心肌成纤维细胞细胞周期和增殖细胞核抗原表达的影响

目的 :研究苦参碱 (Mat)对醛固酮 (Ald)诱导大鼠心肌成纤维细胞 (RCFibs)增殖的影响 ,探讨其抑制心肌纤维化的机制。方法 :利用体外细胞培养技术 ,建立Ald诱导RCFibs纤维化模型 ,通过四氮甲唑蓝 (MTT)比色法测定细胞增殖 ,流式细胞分析仪 (FCM)测定细胞周期及增殖细胞核抗原 (PCNA)的表达、免疫荧光法测定PCNA的表达 ,研究Mat对Ald诱导RCFibs增殖的影响。结果 :Ald +Mat(0 .5mmol·L-1 、0 .2 5mmol·L-1 、0 .1 2 5mmol·L-1 )组与Ald组相比较 ,其细胞增殖抑制率分别为 9.95 6 7%、2 6 .4 6 1 0 %、4 6 .5 90 9% (P <0 .0 5 ) ,细胞周期分析显示G0 G1 期细胞数量增加 ,S期、G2 M1 期明显减少 ;免疫荧光和FCM测定表明 ,Mat抑制PCNA的表达。结论 :Mat能明显抑制Ald诱导的RCFibs增殖 ,其效应呈现剂量依赖性。

Effects of Matrine on Cellcycle and Expression of PCNA of Rat Cardiac Fibroblasts Induced by Aldosterone

Objective: To investigate the effects of matrine(Mat) on the proliferation of rat cardiac fibroblasts(RCFibs) induced by aldosterone(Ald),and to research the mechanism of regression of heart fibrosis. Methods: Through the technique of cell culture in vitro , a model which Ald induced RCFibs fibrosis was established, and prolifertion was measured by thiazolyl blue(MTT) assay. Cellcycle distribution and the expression of PCNA were determined with flow cytometer(FCM),and the expression of proliferation cell nuclear antigen(PCNA) was measured by immunofluorescence staining method to observe the effects of matrine on the prolifertion of RCFibs induced by Ald. Results: Compared with Ald group, Ald+Mat(0.5 mmol·L -1 ,0.25 mmol·L -1 ,0.125 mmol·L -1 )groups could inhibit RCFibs proliferation,the rate of inhibition respectively was 9.9567%,26.4610% and 46.5909% ( P <0.05);Cellcycle distribution analysis indicated that cell number was increased in G 0 G 1 period, and was markedly decreased in S and G 2 M 1 period; Measurement of immunofluorescence method and FCM indicated that Mat markedly inhibited the expression of PCNA. Conclusion: Mat could restrain the proliferation of RCFibs induced by Ald,and this stimulating effect is in a dose dependent manner.