[关键词]
[摘要]
目的 探讨注射用益气复脉(冻干,YQFM)对阿霉素(doxorubicin,DOX)诱导H9c2(2-1)心肌细胞毒性的保护作用。方法 H9c2(2-1)心肌细胞随机分为对照组,模型组(终浓度为0.3 μmol/L的DOX处理细胞48 h),YQFM低、中、高剂量(125、625、3 125 μg/mL)组(提前2 h加入药物预处理,然后加入终浓度为0.3 μmol/L的DOX处理48 h),采用CCK-8法检测细胞活力;使用乳酸脱氢酶(LDH)和ATP试剂盒检测细胞LDH和ATP水平;应用Hoechst 33258染色法检测细胞凋亡;JC-1法检测细胞线粒体膜电位;Western blotting法检测caspase-3蛋白的表达水平。结果 与模型组比较,YQFM中、高剂量组显著增加细胞存活率(P<0.05、0.01),低、高剂量组明显改善细胞凋亡;低、中、高剂量组LDH活性显著降低(P<0.05、0.01),ATP含量显著增加(P<0.05、0.01),线粒体膜电位明显恢复。结论 YQFM抑制DOX诱导H9c2(2-1)的细胞毒性,其作用机制可能与抑制线粒体凋亡信号通路的激活有关。
[Key word]
[Abstract]
Objective To investigate the protective effect of Yiqi Fumai Lyophilized Injection (YQFM) against Doxorubicin (Dox)-induced cardiotoxicity in H9c2 (2-1). Methods H9c2 (2-1) myocardial cells were randomly divided into control group, model group (cells treated with DOX at the final concentration of 0.3 μmol/L for 48 h), YQFM low, medium and high dose (125, 625, and 3 125 g/mL) group (pretreatment with YQFM for 2 h, and then adding DOX of a final concentration of 0.3 μmol/L). Cell viability was detected with CCK-8 assay. Cells LDH and ATP levels were examined using LDH and ATP kits. Cell death was measured by Hoechst 33258 stain assay. The mitochondrial membrane potential was detected by JC-1. Caspase-3 protein expression was evaluated by Western blotting. Results Compared with model group, YQFM medium and high dose group significantly increased the cell survival rate (P < 0.05 and 0.01), low and high dose group significantly improved cell apoptosis; LDH activity of low, medium and high dose group was significantly decreased (P < 0.05 and 0.01), ATP content increased significantly (P < 0.05 and 0.01), and the mitochondrial membrane potential was restored. Conclusion YQFM inhibited DOX-induced cardiotoxicity in H9c2 (2-1), the possible mechanisms may be related to the inhibition of activation of mitochondrial apoptotic signaling pathway.
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[基金项目]
天津市中药注射剂关键技术校企协同创新实验室建设基金(17PTSYJC00090)