睾酮对心肌成纤维细胞表型转化的干预研究.docVIP

摘要】 目的 观察氧化应激对心肌成纤维细胞(CF)表型转化的影响以及睾酮的干预作用，初步探讨睾酮抗心脏老化过程中心肌纤维化的机制。方法 差速贴壁法培养雄性昆明白乳鼠心肌成纤维细胞， 用β半乳糖苷酶瓤色检测细胞的衰老，MTT法检测细胞增殖能力，采用免疫细胞化学染色检测肌成纤维细胞的典型标记α平滑肌肌动蛋白（αSMA）。结果 H2O2刺激下β半乳糖苷酶染色阳性细胞比例增加，心肌成纤维细胞增殖率降低，αSMA免疫化学染色出现明显强阳性；睾酮在3.0×10-9、3.0×10-8和3.0×10-7mol/L 3种浓度下均具有延缓H2O2诱导的以上效应，氟他胺（Flu）能部分阻断睾酮的各种保护作用。结论 睾酮通过受体机制可以逆转H2O2诱导的心脏成纤维细胞表型的转化，这可能是睾酮抗心脏老化过程中心肌纤维化的机制之一。 【关键词】 睾酮;氧化应激；衰老;心肌成纤维细胞;表型 Intervention effect of testosterone on phenotype of cardiac fibroblasts induced by oxidative stress LIU XinQiang, WU SaiZhu, XING XiaoWen,et al. Department of Cardiology, Nanfang Hospital Affiliated to Southern Medical University, Guangzhou 510515, Guangdong, China 【Abstract】 Objective To observe the intervention effect of testosterone on phenotype of cardiac fibroblasts induced by oxidative stress. Methods Cardiac fibroblast from ventricle of heart of neonatal male Kunming mice were cultured by differential anchoring velocity method. The expression of SAβgal in each group was detected through senescenceassociated βgal staining and the proliferation of cells was detected by MTT method. Protein level of αsmooth muscle actin (αSMA) which was the typical mark of myofibroblast was measured by immunostaining. Results Compared to control group, H2O2 caused more cellular senescence and depressed the proliferation of cells, and the expression of αSMA was significantly stimulated. Under these concentrations(3×10-9, 3×10-8 and 3×10-7mol/L), testosterone pretreatment of cells significantly attenuated all these effects of H2O2 exposure. Flutamide administered prior to testosterone(3×10-8 mol/L) pretreatment and partly antagonized the protective effects of testosterone against H2O2 exposure.Conclusions The reversing effects of testosterone on the phenotype conversion of cardiac fibroblasts mediated by the androgen receptors might be one of the mechanisms for fighting against cardiac fibrosis during aging process of heart. 【Key words】 Testosterone; Oxidative