【摘要】 目的探讨苦参碱对醛固酮诱导的新生大鼠心肌成纤维细胞增殖、胶原代谢的影响,并探讨转化生长因子-β1(TGF-β1)信号转导通路在苦参碱抗心肌纤维化中的作用机制。方法体外差速贴壁法分离纯化培养心肌成纤维细胞,试验分为对照组、醛固酮(1.0×10-7mol/L)组和醛固酮+不同浓度(0.125,0.25,0.5mmol/L)苦参碱组5组。作用24h后,倒置显微镜观察细胞生长状态;四甲基偶氮唑盐(MTT)法检测心肌成纤维细胞增殖;流式细胞分析仪分析细胞周期;分光光度法检测细胞上清羟脯氨酸含量;细胞免疫荧光染色及流式细胞仪检测TGF-β1蛋白表达。结果与对照组比较,醛固酮可显著提高心肌成纤维细胞MTT-OD值,使S期细胞比率增加,G0/G1期细胞比率下降;羟脯氨酸含量、TGF-β1的蛋白表达均显著高于对照组(P<0.01)。苦参碱干预24h后,MTT-OD值、S期的细胞百分比、羟脯氨酸含量、TGF-β1的蛋白表达均显著低于醛固酮组(P<0.01),且苦参碱的效应呈浓度依赖性。结论苦参碱可抑制醛固酮诱导的心肌成纤维细胞增殖及胶原合成,其抗纤维化作用可能与直接下调TGF-β1蛋白表达有关。更多还原

【Abstract】 Objective To investigate the effects of matrine(Mat) on the proliferation and collagen synthesis of rat cardiac fibroblasts(CFs) induced by aldosterone(Ald),and to research the role of transforming growth factor beta(TGF-β1) in the regression of heart fibrosis.Methods The neonatal rat cardiac fibroblasts were extracted by enzymatic digestion and in vitro different velocity adherence method.Through the technique of cell culture in vitro,to establish the model CFs fibrosis induced by Ald,then to add disposal factors.The cardiac fibroblasts were divided into 5 groups:normal control group,Ald(1.0 × 10-7 mol /L) group,Ald + Mat(0.125,0.25,0.5 mmol /L) groups.After 24 h action,proliferation was measured by thiazolyl blue(MTT) assay;cell cycle distribution was determined with flow cytometer(FCM);the expression of TGF-β1 was measured by immunofluorescence staining method and FCM.Collagen synthesis was determined by means of hydroxyproline concentration staining.Results Compared with the normal control group,Ald groups markedly enhanced CFs.MTT results and the expression of TGF-β1,cell cycle distribution analysis indicated that Ald groups’ cell numbers were decreased in G0 /G1 period,markedly increased in S period.Ald greatly enhanced collagen synthesis content and the expression of TGF-β1.Compared with Ald groups,Ald + Mat groups markedly decreased CFs.MTT results,cell cycle distribution analysis indicated that Ald + Spi groups’ cell numbers were increased in G0 /G1 period,markedly decreased in S period.The expression of TGF-β1 was decreased.The inhibition effect was of dose dependent-manner.Conclusion Mat could effectively inhibit the proliferation of Ald induced CFs by downregulating TGF-β1 protein expression in a concentration dependent manner,which may play an important role in the regression of heart remodeling.更多还原