苦参素对缺氧/复氧损伤乳鼠心肌细胞氧化应激和细胞凋亡的影响  

The Influence of Oxymatrine on Oxidative Stress and Apoptosis in Neonatal Rat Impaired by Hypoxia-reoxygenation

作　　者：陈树杰[1] 杨晓龙[1] 王雪芬[2] 陈晨[1] 王朝宗[1] 

机构地区：[1]河北省邯郸市中心医院,河北邯郸056001 [2]河北省邯郸市第二医院

出　　处：《中西医结合心脑血管病杂志》2016年第20期2366-2371,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河北省卫生厅重点科技研究计划(No.20130359)

摘　　要：目的研究苦参素(OMT)对缺氧/复氧损伤乳鼠心肌细胞氧化应激和细胞凋亡的影响。方法无菌环境下分离SD大鼠乳鼠心肌细胞并培养72 h后随机分为:空白对照组、缺氧/复氧(H/R)模型对照组、OMT(20μg/m L、40μg/m L和80μg/m L)干预组和舒血宁注射液(SXN,100μg/m L)干预组(阳性对照组),每组设10个复孔。药物干预6 h后,通过MTT法检测细胞存活率;检测细胞培养液中心肌酶[谷草转氨酶(AST)、磷酸肌酸激酶(CPK)、乳酸脱氢酶(LDH)]释放量;测定细胞中抗氧化酶[超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)]活性和丙二醛(MDA)含量;通过流式细胞仪检测细胞凋亡状况并计算凋亡率,通过RT-PCR检测细胞中bcl-2mRNA、BaxmRNA表达并计算bcl-2/Bax比值,通过Western blot方法检测细胞caspase-3蛋白表达水平并进行半定量分析。结果与模型对照组比较,OMT40μg/m L、80μg/m L干预组、SXN干预组乳鼠心肌细胞存活率显著升高,培养液中AST、CPK、LDH释放量显著降低,细胞中SOD、CAT活性显著升高且MDA含量显著降低,细胞凋亡状况显著改善、凋亡率显著降低,细胞中bcl-2 mRNA表达显著上调且Bax mRNA表达显著下调、bcl-2/Bax表达比值显著升高,细胞中caspase-3蛋白表达显著升高,上述差异均有统计学意义(P<0.05或P<0.01);其中OMT 80μg/m L干预组细胞中GSH-Px活性显著升高(P<0.05)。结论苦参素对缺氧/复氧损伤乳鼠心肌细胞氧化应激和细胞凋亡具有抑制作用,其作用机制可能与苦参素能有效改善抗氧化酶活性、调节凋亡相关基因表达有关。Objective To investigate the effects of oxymatrine（ OMT） on oxidative stress and apoptosis in neonatal rat impaired by hypoxia-reoxygenation（ H / R）. Methods Cadiocytes of neonate rats were cultivated for 72 hours and divided into six groups： normal control group,H /R model control group,OMT（ 20 μg / m L,40 μg / m L and 80 μg / m L） treatment groups and Shuxuening injection（ SXN,100 μg / m L） treatment group（n = 10）. After 6 hours of drugs intervention,the survival rate was detecd. The content of aspartate transaminase（ AST）,creatine phosphokinase（ CPK）,lactate dehydrogenase（ LDH） in culture medium were deteced. The activity of superoxide dismutase（ SOD）,catalase（ CAT）,glutathione peroxidase（ GSH-Px） and the content of malondialdehyde（ MDA） in cardiomyocytes were determined. The apoptosis was observed by flow cytometry and the apoptosis rate was calculated. The expression of bcl-2 mRNA,Bax mRNA were detected by reverse transcription polymerase chain reaction（ RT-PCR）. The ratio of bcl-2 / Bax was calculated. The expression of caspase-3 protein was detected by western blot. Results Compared with the model control group,the survival rate were significantly increased in OMT（ 40 μg / m L,80 μg /m L） treatment groups. The content of CPK,AST,LDH in culture medium were significantly decreased. The activity of SOD,CAT in cardiomyocytes were significantly increased and the content of MDA was significantly decreased. The apoptosis rate was significantly decreased. The expression of bcl-2 was significantly up-regulated,the expression of Bax and caspase-3 were significantly down-regulated. The ratio of bcl-2 / Bax were significantly increased. The activity of GSH-Px was significantly increased（P〈0. 05）. All of the difference above were significant（P〈0. 05 or P〈0. 01）. Conclusion OMT had inhibitive effects on oxidative stress and apoptosis of neonatal rat impaired by H / R,which perhaps related to its effects of improving the activity of antioxidase

关 键 词：缺氧/复氧损伤 心肌细胞 苦参素 氧化应激 细胞凋亡 

分 类 号：R259[医药卫生—中西医结合]