积雪草酸对新生大鼠心肌细胞缺血-再灌注损伤的保护作用  

Protective effect of asiatic acid on myocardial ischemia-reperfusion injury in newborn rats

作　　者：易晨龙 许晶 司林杰 张郁青[2] 王晓伟 

机构地区：[1]南京医科大学第一附属医院心胸外科,江苏省210029 [2]南京医科大学附属江宁医院心血管病科

出　　处：《江苏医药》2015年第15期1743-1746,F0003,共5页Jiangsu Medical Journal

基　　金：江苏省中医局项目(LZ13127);江苏省教育厅自然科学基金(12KJB320003)

摘　　要：目的探讨积雪草酸(AA)对氧糖剥夺/复氧模型模拟的新生大鼠心肌细胞缺血-再灌注(I-R)损伤的保护作用。方法将体外培养原代新生大鼠心肌细胞分为四组:D组继续正常培养;A、B和C组分别采用生理盐水、<0.1%二甲基亚砜溶液和AA 20μmol/L培养24h后,采用氧糖剥夺处理6h-复氧12h建立I-R损伤模型。采用TUNEL法测定心肌细胞凋亡率;流式细胞术检测细胞内活性氧簇(ROS)荧光强度;免疫荧光显微镜观察心肌细胞内线粒体膜电位;Western blot法检测心肌细胞线粒体凋亡通路相关蛋白及丝裂原活化蛋白激酶(MAPK)信号转导通路相关蛋白。结果A、B和C组心肌细胞凋亡率、ROS水平、低线粒体膜电位的心肌细胞比例、B淋巴细胞瘤-2基因相关X蛋白/B淋巴细胞瘤-2基因比例以及细胞色素C、磷酸化p38、磷酸化细胞外信号调节激酶1/2和磷酸化c-Jun氨基末端激酶表达均高于D组(P<0.05),而C组上述指标低于A组(P<0.05)。结论 AA对氧糖剥夺/复氧模型诱导的心肌细胞I-R损伤有保护作用,可抑制ROS诱导的心肌细胞凋亡;其机制可能与抑制线粒体凋亡通路的活化及阻滞MAPK信号激酶磷酸化有关。Objective To explore the protective effect of asiatic acid （AA） on myocardial ischemia-reperfusion（I-R） injury in newborn rats simulated by oxygen-glucose deprivation （OGD）/ reoxygenation. Methods Myocardial cells of newborn rats were cultured in vitro and divided into four groups. The cells of group D were cultured continuously as blank controls. The cells of groups of A,B and C were cultured with normal saline,〈0. 1% DMSO and AA 20μmol/L, respectively, for 24 hours,and then treated with OGD for 6 hours and reoxygenation for 12 hours. Cell apoptosis was examined by TUNEL method, fluorescence intensity of ROS was determined by flow cytometry, mitochondrial membrane potential （MMP） was measured by immunofluorescence microscopy, and myocardial mitochondrial apoptotic pathways-related proteins and MAPK signal transduction pathway-related proteins were determined by Western blot. Results Myocardial cells apoptosis rate, ROS level,the proportions of low MMP of cardiac cells, the Bax/Bcl-2 ratio and the expressions of Cyt-C, phosphorylation-p38, phosphorylation-ERK1/2 and phosphorylation-JNK were all higher in groups of A,B and C than those in group D（P〈0. 05）, which were lower in group C than those in group A（P〈0. 05）. Conclusion AA has a protective effect on the myocardial I-R injury and an inhibitory effect on the myocardial apoptosis, which may be related to the inhibition of the mitochondrial apoptosis pathway activation and retardation of MAPK signal kinase phosphorylation.

关 键 词：积雪草酸 心肌缺血-再灌注损伤 

分 类 号：R542[医药卫生—心血管疾病]