Oxymatrine inhibits aldosterone-induced rat cardiac fibroblast proliferation and differentiation by attenuating smad-2,-3 and-4 expression: an in vitro study.
Oxymatrine inhibits aldosterone-induced rat cardiac fibroblast proliferation and differentiation by attenuating smad-2,-3 and-4 expression: an in vitro study.
复制标题
DOI：
10.1186/s12906-016-1231-9
复制
发表时间：
2016-07-26
影响因子：
--
通讯作者：
Shen X
中科院分区：
文献类型：
--
作者：
Fu L;Xu Y;Tu L;Huang H;Zhang Y;Chen Y;Tao L;Shen X
我们之前证明氧化苦参碱（一种中药苦参中的生物碱）可以改善血流动力学紊乱和心脏纤维化；然而，其根本机制尚不清楚。在这里，我们研究了氧化苦参碱对醛固酮诱导的体外心肌成纤维细胞向肌成纤维细胞分化的影响和作用机制。心脏成纤维细胞是从新生斯普拉格道利大鼠中分离纯化的。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑 (MTT) 测定确定刺激心脏成纤维细胞增殖的醛固酮的最佳浓度。心脏成纤维细胞用7.57 × 10−4 mol/L或3.78 × 10−4 mol/L氧化苦参碱或不加氧化苦参碱预处理2 h，然后与1 × 10−8 mol/L醛固酮共孵育48 h。 MTT法和Masson染色检测心肌成纤维细胞增殖和肌成纤维细胞分化。采用市售ELISA试剂盒测定I型和III型胶原蛋白的分泌，并通过比色法测定羟脯氨酸含量。 Western blotting 检测心脏成纤维细胞中 Smad-2、Smad-3 和 Smad-4 蛋白的表达。 目前的结果证实，醛固酮诱导心脏成纤维细胞向肌成纤维细胞增殖和分化。 MTT法和Masson染色表明氧化苦参碱显着抑制醛固酮诱导的心肌成纤维细胞增殖和肌成纤维细胞分化。氧化苦参碱显着抑制醛固酮诱导的 I 型和 III 型胶原蛋白分泌（如商业 ELISA 试剂盒所示），以及醛固酮诱导的羟脯氨酸含量增加（如比色测定所示）。蛋白质印迹显示氧化苦参碱可减弱醛固酮诱导的心脏成纤维细胞中的 Smad-2、Smad-3 和 Smad-4 表达。氧化苦参碱可通过与减弱 Smad 信号通路相关的机制抑制心脏成纤维细胞增殖和分化为肌成纤维细胞。
We previously demonstrated oxymatrine, an alkaloid from the Chinese medicine radix Sophorae flavescentis, ameliorates hemodynamic disturbances and cardiac fibrosis; however, the underlying mechanisms are unclear. Here, we investigated the effect and mechanism of action of oxymatrine on aldosterone-induced cardiac fibroblast to myofibroblast differentiation in vitro. Cardiac fibroblasts were isolated purified from neonatal Sprague Dawley rats. The optimal concentration of aldosterone to stimulate cardiac fibroblast proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cardiac fibroblasts were pretreated with 7.57 × 10−4 mol/L or 3.78 × 10−4 mol/L oxymatrine or without oxymatrine for 2 h, and then coincubated with 1 × 10−8 mol/L aldosterone for 48 h. The MTT assay and Masson staining were used to detect the cardiac fibroblast proliferation and myofibroblast differentiation. The secretion of type I and III collagen was measured by commercial ELISA kits, and the hydroxyproline content was determined by the colorimetric assay. Western blotting assayed the Smad-2, Smad-3, and Smad-4 protein expression in cardiac fibroblasts. The present results confirmed that aldosterone induced cardiac fibroblast to myofibroblast proliferation and differentiation. The MTT assay and Masson staining indicated oxymatrine significantly inhibited aldosterone-induced cardiac fibroblast proliferation and myofibroblast differentiation. Oxymatrine significantly inhibited aldosterone-induced secretion of type I and III collagen, as indicated by commercial ELISA kits, and aldosterone-induced increase in hydroxyproline content, as indicated by a colorimetric assay. Western blotting revealed oxymatrine attenuated aldosterone-induced Smad-2, Smad-3, and Smad-4 expression in cardiac fibroblasts. Oxymatrine can inhibit cardiac fibroblast proliferation and differentiation into myofibroblasts via a mechanism linked to attenuation of the Smad signaling pathway.