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目的:基于细胞水平研究小檗碱(berberine,Ber)对阿霉素(doxorubicin,DOX)所致心肌损伤的保护作用并探讨其抗氧化机制。方法:大鼠心肌细胞H9c2给药组分为:空白组,2μmol·L~(-1)DOX组,0.1μmol·L~(-1)Ber组,1μmol·L~(-1)Ber组,10μmol·L~(-1)Ber组,2μmol·L~(-1)DOX+0.1μmol·L~(-1)Ber组,2μmol·L~(-1)DOX+1μmol·L~(-1)Ber组,2μmol·L~(-1)DOX+10μmol·L~(-1)Ber组。各组给药不同时间后测定细胞体积及蛋白含量,RT-PCR法测定细胞心钠素(ANP),脑钠素(BNP)表达水平。以半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性为指标评价心肌细胞凋亡水平。DCFH荧光探针法测定细胞ROS水平,同时测定细胞内丙二醛(MDA)生成量,评价心肌细胞氧化损伤水平。结果:与空白组比较,各给药组心肌细胞体积增大,细胞内蛋白含量增加,ANP,BNP的mRNA水平上调,细胞内Caspase-3活性增强,导致细胞凋亡,降低细胞内ROS水平,均具有明显的统计学差异(P<0.01)。结论:小檗碱通过抗氧化作用降低阿霉素诱导的心肌细胞损伤。小檗碱与阿霉素合用后,浓度依赖的抑制阿霉素诱导的心肌肥大,并有效降低细胞内ROS水平,清除脂质过氧化物,逆转阿霉素诱导的心肌细胞凋亡。
OBJECTIVE: To study the protective effect of berberine (Berberine) against myocardial injury induced by doxorubicin (DOX) and to explore its anti-oxidative mechanism based on cellular level. Methods: The H9c2-administered rat myocardial cells were divided into blank group, 2μmol·L -1 DOX group, 0.1μmol·L -1 Ber group, 1μmol·L -1 Ber group, 2μmol·L -1 DOX + 1μmol·L -1 Ber 2 group, 2μmol·L -1 DOX + 0.1μmol·L -1 Ber group, 2μmol·L -1 DOX + 1μmol·L -1 ) Ber group, 2μmol·L -1 DOX + 10μmol·L -1 Ber group. The cell volume and protein content were determined at different time points after administration. The expression of ANP and BNP was determined by RT-PCR. The apoptosis of cardiomyocytes was evaluated by Caspase-3 activity. DCFH fluorescent probe method was used to determine the level of ROS in the cells. Meanwhile, the amount of intracellular malondialdehyde (MDA) was measured and the level of oxidative damage of cardiomyocytes was evaluated. Results: Compared with the blank group, the volume of myocardial cells increased, the content of intracellular protein increased, the mRNA level of ANP and BNP increased, the activity of Caspase-3 increased, which led to the apoptosis of cells and the decrease of intracellular ROS levels. There was a significant statistical difference (P <0.01). Conclusion: Berberine can reduce doxorubicin - induced cardiomyocyte injury through anti - oxidative effect. Berberine combined with doxorubicin, a concentration-dependent inhibition of doxorubicin-induced cardiac hypertrophy, and effectively reduce intracellular ROS levels, clear lipid peroxides, reverse doxorubicin-induced cardiomyocyte apoptosis.