通心络对高糖培养下心肌成纤维细胞增殖和凋亡的影响

Effects of Tongxinluo on High Glucose-induced Proliferation and Apoptosis of Cardiac Fibroblasts

目的观察通心络对高糖培养下心肌成纤维细胞增殖和凋亡的影响,探讨其抑制心肌纤维化的可能机制。方法原代和传代培养SD大鼠乳鼠的心肌成纤维细胞,并用免疫荧光染色法鉴定。细胞分为对照组(低糖DMEM培养)、模型组(高糖DMEM培养)和通心络处理组(高糖DMEM+通心络20、80、320μg/m L培养)。MTT检测各组细胞的增殖活力;ELISA检测各组Ⅰ、Ⅲ型胶原蛋白的分泌;Western blotting检测各组bax和bcl-2蛋白的表达。结果模型组细胞的增殖活力及分泌的Ⅰ、Ⅲ型胶原蛋白均高于对照组,而通心络处理组细胞的增殖活力和分泌的胶原蛋白明显低于模型组;与对照组相比,模型组的bcl-2表达明显增多,bax表达减少;与模型组相比,通心络处理组的bcl-2表达减少,bax表达增加。结论通心络可以减弱高糖培养下心肌成纤维细胞的增殖,并抑制其过度分泌Ⅰ、Ⅲ型胶原蛋白,同时还可通过减少bcl-2、增加bax的表达诱导心肌成纤维细胞凋亡,从而发挥其治疗心肌纤维化的作用。

Objective This study investigated the effect of tongxinluo （TXL） on high glucose-induced proliferation and apoptosis of cardiac fibroblasts （CFs） ,to explore the possible mechanism by which TXL inhibits myocardial fibrosis. Methods Primary culture and subculture of neonatal SD rat CFs was carried out as follows. Immunofluorescence staining was performed to identify CFs. The CFs were divided into control group （cultured with low-glucose DMEM） ,model group （cultured with high-glucose DMEM） , and TXL treatment groups （cultured with high-glucose DMEM＋TXL at 20, go, and 320 μg/mL）. The proliferation of CFs in each group was detected by MTT assay. The expression of collagen types Ⅰ and Ⅲ in each group was detected by ELISA. Western blotting was used to detect the expression of bax and bel-2 in each group. Results CFs Proliferation and collagen types I andⅢsecretion were higher in the model group than in the control group. The CFs proliferation and collagen content in the TXL treatment groups were significantly lower than those in the model group, hcl-2 expression was significantly increased and hax expression was decreased in the model group, compared with the corresponding values in the control group. In comparison with the model group, bcl-2 expression was downregulated and bax expression was upregulated in the TXL treatment groups. Conclusion TXL can reduce the CFs proliferation and inhibit their collagen secretion under high glucose conditions. TXL also induces the CFs apoptosis by reducing bcl-2 expression and increasing bax expression. Thus,TXL can play a role in the treatment of myocardial fibrosis.