广枣总黄酮对三氧化二砷诱导心肌细胞损伤的作用

Effect of total flavonoids of fructus choerospondiatis on arsenic trioxide-induced cardiomyocyte injury

目的研究广枣总黄酮(TFFC)对三氧化二砷(ATO)诱导心肌细胞损伤的影响,并探讨其作用机制。方法分离并培养乳鼠心肌细胞,倒置显微镜下观察不同时间段的心肌细胞形态。用6μg/ml ATO诱导建立原代心肌细胞损伤模型,实验设置空白组,模型组,TFFC低、中、高浓度组,采用CCK-8法检测各组细胞存活率,全自动生化分析仪检测细胞培养液中谷草转氨酶(GOT)、磷酸肌酸激酶(CK)、乳酸脱氢酶(LDH)的活力,试剂盒法检测细胞中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)以及过氧化氢酶(CAT)的活力,Hoechst 33342荧光染色检测细胞凋亡程度,流式细胞术定量测定细胞凋亡率。结果培养48 h的细胞基本全部贴壁,大多数细胞伸出伪足并伴有自发性搏动,可满足实验要求,6μg/ml ATO可降低细胞存活率。与模型组比较,TFFC各浓度组细胞存活率显著升高,细胞培养液中LDH、CK、GOT的活力显著降低,细胞中SOD、GSH-Px、CAT的活力显著升高,细胞凋亡率显著降低,差异有统计学意义(P<0.05)。结论TFFC可改善ATO诱导心肌损伤状态,从而对ATO损伤的心肌起到保护作用。

Objective To study the effect of total flavnoids of fructus choerospondiatis(TFFC)on arsenic trioxide(ATO)-induced cardiomyocyte injury,and to explore its mechanism of action.Methods Cardiomyocytes were isolated and cultivated by neonatal rats and the morphology of cardiomyocytes at different time periods was observed under an inverted microscope.Cardiomyocyte injure models were use established by 6μg/ml ATO,the experiment set up blank group,model group,and TFFC low,medium,and high concentration groups.The CCK-8 method was used to detect the survival rate of each group of cells,the activity of glutamic oxaloacetic transaminase(GOT),creatine kinase(CK)and lactate dehydrogenase(LDH)in cell culture medium were determined by automatic bio-chemical instrument;the activity of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)in the cells were detected by kit method.The degree of apoptosis was observed by Hoechst 33342 fluorescence staining.Cell apoptosis rates were measured by flow cytomety.Results The cells cultured for 48 hours were basically all attached to the wall.Most of the cells stuck out the pseudopodia and were accompanied by spontaneous beating,which can meet the experimental requirements.The cell survival rate was reduced by 6μg/ml ATO.Compared with the model group,the cell survival rate of each TFFC concentration group significantly increased,the vitality of LDH,CK,and GOT in the cell culture fluid was significantly reduced,the vitality of SOD,GSH-Px,and CAT in the cell were significantly increased,and the apoptosis rate was significantly reduced,and the differences were statistically significant(P<0.05).Conclusion TFFC can improve the state of ATO-induced myocardial injury,thereby protecting the myocardium damaged by ATO.