参麦注射液对内毒素诱导心肌细胞凋亡的抑制作用及细胞Wnt2/β-catenin表达的影响

Inhibitory Effect of Shenmai Injection on Cardiomyocyte Apoptosis Induced by Endotoxin and Effect of Wnt2/β-catenin Expression

为探讨参麦注射液诱导心肌细胞凋亡的作用机理,本研究分别选取对照组、内毒素（LPS）处理组、参麦注射液（SMI）＋内毒素（LPS）处理组,共3组小鼠心肌细胞进行试验。通过MTT检测了各实验组的心肌细胞存活率,利用流式细胞仪检测心肌细胞凋亡数量,最后利用RT-PCR和Western blotting分别检测心肌细胞Wnt2/β-catenin m RNA的表达和细胞内Wnt2/β-catenin蛋白的表达。结果显示SMI进行剂量干预后,内毒素诱导心肌细胞存活率显著升高（p〈0.05）。当SMI的干预量为0.8 g/L时,细胞存活率高于LPS组（p〈0.05）;SMI以1 g/L剂量进行干预时,细胞存活率显著高于LPS组（p〈0.01）。同时,SMI干预的心肌细胞中,UR、LR区的细胞比例较LPS组有显著下降（p〈0.05）。SMI干预内毒素诱导心肌细胞后,显著逆转内毒素诱导心肌细胞内Wnt2/β-catenin m RNA的表达（p〈0.05）,并且Wnt2/β-catenin蛋白的表达也得到显著性逆转（p〈0.05）。该结果表明参麦注射液主要是通过影响心肌细胞Wnt2/β-catenin信号通路,从而抑制内毒素诱导的心肌细胞的凋亡。

To explore the mechanism of Shenmai injection in inducing cardiomyocyte apoptosis, the control group, endotoxin（LPS） treatment group, Shenmai injection（SMI） ＋ endotoxin（LPS） treatment group were selected in this study, a total of three groups of small Rat cardiomyocytes were tested. The survival rate of cardiomyocytes in each experimental group was measured by MTT. The number of myocardial cell apoptosis was detected by flow cytometry. The expression of Wnt2/β-catenin m RNA and intracellular Wnt2 were detected by RT-PCR and Western blot respectively. Expression of β-catenin protein. The results showed that the endotoxin-induced cardiomyocyte survival rate significantly increased after SMI dose intervention（p〈0.05）. When SMI was treated with0.8 g/L, the cell survival rate was significantly higher than that of LPS group（p〈0.05）. When SMI was treated with1 g/L dose, the cell survival rate was significantly higher than that of LPS group（p〈0.01）. At the same time, the ratio of UR and LR cells in SMI-treated cardiomyocytes was significantly lower than that in LPS group（p〈0.05）. After intervention with endotoxin-induced cardiomyocytes, SMI significantly reversed the expression of Wnt2 andβ-catenin m RNA in cardiomyocytes induced by endotoxin（p〈0.05）, and the expression of Wnt2 and β-catenin protein was also significantly reversed（p〈0.05）. The results indicate that Shenmai injection mainly affects the Wnt2/β-catenin signaling pathway of cardiomyocytes, thereby inhibiting endotoxin-induced cardiomyocyte apoptosis.