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目的研究清心饮对柯萨奇B_3病毒(Coxsackievirus B_3,CVB_3)感染心脏微血管内皮细胞(cardiac microvascular endothelial cells,CMVECs)的保护作用。方法采用RT-PCR检测CVB_3 RNA,应用倒置显微镜观察CMVECs形态,CCK8检测细胞活性,选择清心饮最佳干预时间点和稀释浓度。在最佳条件下通过免疫荧光法、Western blot法检测内皮细胞标志物CD31,间充质细胞标志物a-SMA的表达。结果 CVB_3能在CMVECs中持续性复制,清心饮最佳干预时间点和浓度分别是72 h和20%含药血清。与空白组比较,CVB_3组CMVECs形态固缩、变异,CD31表达减弱,a-SMA表达增强;与空白血清组比较,清心饮血清组CMVECs形态趋于正常,CD31表达增强,a-SMA表达减弱。结论清心饮可抑制CVB_3感染CMVECs发生内皮细胞转分化,对CMVECs具有保护作用。
Objective To study the protective effect of Qingxin Decoction on cardiac microvascular endothelial cells (CMVECs) infected with Coxsackievirus B_3 and CVB_3. Methods CVB 3 RNA was detected by RT-PCR. The morphological changes of CMVECs were observed by inverted microscope. The cell viability was detected by CCK8. The optimal time and concentration of Qingxin Decoction were selected. Under the optimal conditions, the expression of endothelial cell marker CD31 and mesenchymal cell marker a-SMA were detected by immunofluorescence and Western blot. Results CVB_3 could be replicated continuously in CMVECs. The optimal time and concentration of Qingxin Decoction were 72 h and 20% serum respectively. Compared with the blank group, the CMVECs of CVB_3 group were shrinkage, the expression of CD31 was decreased and the expression of a-SMA was increased. Compared with the blank serum group, CMVECs in normal serum group tended to be normal, CD31 expression was decreased and a-SMA expression was decreased. Conclusions Qingxin Decoction can inhibit the endothelial cell transdifferentiation induced by CVB_3 infection in CMVECs and has a protective effect on CMVECs.