靶向沉默HDAC6对乳鼠心房肌成纤维细胞活化增殖的作用

目的 探究组蛋白去乙酰化酶(HDAC6)对乳鼠心房肌成纤维细胞活化增殖的影响作用.方法 将50只乳鼠(雄性)分别提取心房肌成纤维细胞,应用Western blot法测定HDAC6和α-平滑肌肌动蛋白(α-SMA)的表达情况;运用qRT-PCR技术检测HDAC6和α-SMA mRNA的表达情况;MTT法检测心房肌成纤维细胞的细胞增殖活性;使用LipofectamineTM 2000 Reagent向心房肌成纤维细胞中转染小干扰RNA HDAC6(siRNA-HDAC6)后观察对心房肌成纤维细胞增殖的影响.结果 转染心房肌成纤维细胞siRNA-HDAC6组HDAC6和α-SMA的蛋白表达明显下降;qRT-PCR技术检测siRNA-HDAC6组中HDAC6和α-SMA的mRNA表达明显下降;转染siRNA-HDAC6明显抑制心房肌成纤维细胞的增殖活性.结论 靶向沉默HDAC6可抑制心房肌成纤维细胞活化增殖,提示其在心房纤维化中可能发挥调控作用.

The effect of HDAC6 silencing onproliferation in neonatal rat atrial fibroblasts

Objective To explore influence of histone deacetylase 6(HDAC6) on neonatal rat atrial fibroblasts activation and proliferation.Methods Atrial fibroblasts were treated with transforming growth factor-β1(TGF-β1) for 48 hours.qRT-PCR was applied to assess the expression of HDAC6 and α-smooth muscle actin(α-SMA) mRNA levels.The protein expression levels of HDAC6 and α-SMA were detected by Western blot.MTT assay was used to determine the proliferation of atrial fibroblasts.LipofectamineTM 2 000 Reagent was used to transfected atrial fibroblasts with siRNA-HDAC6.Results After cardiac fibroblasts transfected with siRNA-HDAC6, HDAC6 expression was declined and α-SMA expression was decreased.The atrial fibroblasts treated with TGF-β1 that HDAC6 expression was increased and α-SMA expression was increased.The activity of cardiac fibroblasts was decreased after transfected with siRNA-HDAC6.Conclusion HDAC6 can suppress proliferative activity of atrial fibroblasts markedly and could be a new therapeutic target of preventing atrial remodeling.The application of its regulator will provide the new idea and method for the prevention and cure of atrial fibrosis.