芍药苷对阿霉素诱导心肌细胞凋亡的保护作用及机制研究

Protective Effect and Mechanism of Paeoniflorin on Doxorubicin-Induced Cardiomyocyte Apoptosis in Rats

目的:探讨芍药苷对阿霉素诱导大鼠心肌细胞凋亡的保护作用及其潜在的机制。方法:雄性SD大鼠60只,随机分成正常对照组、模型对照组、芍药苷10、20、40 mg/kg组和右丙亚胺150 mg/kg组,每组10只。除正常对照组外,大鼠尾静脉给予阿霉素15 mg/kg以诱导大鼠心肌细胞凋亡模型。芍药苷在静脉注射阿霉素前灌胃给药,连续3 d。给药结束后,分离血清和心脏。HE染色观察心肌组织形态学的改变;比色法测定血清中乳酸脱氢酶(LDH)和肌酸激酶(CK)的浓度;TUNEL染色检测心肌细胞凋亡;DCFH-DA荧光法检测心肌组织中活性氧(ROS)的水平;Western blot检测心肌组织线粒体和细胞浆中细胞色素c蛋白的表达;Real-time PCR和比色法分别检测心肌组织中半胱氨酸蛋白酶-3(Caspase-3)的mRNA表达和活性。结果:与正常对照组比较,模型对照组的心肌组织形态学产生了明显的改变,血清LDH和CK浓度、心肌细胞凋亡率、ROS水平、线粒体细胞色素c的释放,以及caspase-3 mRNA表达和活性均明显升高(P<0.05或P<0.01);与模型对照组比较,芍药苷20、40 mg/kg可明显抑制阿霉素诱导的上述效应(P<0.05或P<0.01)。结论:芍药苷对阿霉素诱导的心肌细胞凋亡具有保护作用,其机制可能与抑制ROS的产生进而减少线粒体细胞色素c的释放,从而下调casapse-3的表达和活性有关。

Objective:To investigate the protective effects of paeoniflorin(PEF)on doxorubicin(DOX)-induced cardiomyocyte apoptosis in rats and its potential mechanism.Methods:60 SD male rats were randomly divided into six groups:the control group,the model group,PEF groups(10,20,40 mg/kg),and Dexrazoxane(Dexra 150 mg/kg)group,10 rats in each group.Except for the control group,all other rats were injected with DOX(15 mg/kg)via the tail vein to establish the cardiomyocyte apoptosis model.PEF was administered intragastrically for 3 days before intravenous injection of DOX.At the end of drugs administration,the serum and hearts were collected.The morphological changes of myocardial tissues were observed by HE staining.The concentrations of lactate dehydrogenase(LDH)and creatine kinase(CK)in serum were detected by colorimetry.The apoptosis of myocardial cells was detected by TUNEL.The levels of the reactive oxygen species(ROS)in myocardial tissues were determined by DCFH-DA fluorescence assay.The protein expression of cytochrome c in mitochondria and cytosol of myocardial tissues were detected by Western blot.The Caspase-3 mRNA expression and activity were measured by real-time PCR and colorimetric assay.Results:Compared with the control group,the myocardial histomorphology of the model rats had significant changes,the concentrations of LDH and CK,cardiomyocyte apoptosis rate,ROS level,mitochondrial cytochrome c release,and caspase-3 mRNA expression and activity were significantly increased in the model group(P<0.05 or P<0.01).Compared with the model group,20 and 40 mg/kg paeoniflorin significantly inhibited the above effects induced by DOX.Conclusion:PEF has protective effects on DOX-induced cardiomyocyte apoptosis,which may be related to inhibition of mitochondrial cytochrome c release by decreasing ROS production,to down-regulate the expression and activity of casapse-3.