苦参碱对柯萨奇B_3型病毒感染心肌细胞的保护作用及其与磷脂酰肌醇3-激酶/蛋白激酶B通路的关系  

Protective Effect of Matrine on Cardiomyocytes Infected with Coxsackie Virus B_3 and Its Relationship with Phosphatidylinositol 3-Kinase/Protein Kinase B Singnal Pathway

作　　者：孙永梅[1] 初桂兰[1] 陈霞[2] 赵红[1] 

机构地区：[1]天津医科大学总医院儿科,天津300052 [2]吉林大学基础医学院药理教研室,长春130021

出　　处：《实用儿科临床杂志》2010年第18期1428-1431,共4页Journal of Applied Clinical Pediatrics

摘　　要：目的探讨苦参碱对柯萨奇B3型病毒(CVB3)感染心肌细胞的保护作用及其与磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/AKT)信号传导通路的关系。方法通过CVB3感染原代培养的Wistar新生大鼠心肌细胞的方法,制备病毒性心肌炎体外实验模型。实验分为5组,正常对照组:不感染病毒、不加苦参碱及PI3K抑制剂;病毒组:感染病毒、不加苦参碱及抑制剂Wort;病毒+Wort组:加入抑制剂Wort(200 nmol.L-1)后感染病毒;苦参碱治疗组:感染病毒后再加入苦参碱(0.3 g.L-1);苦参碱治疗+Wort组加入抑制剂Wort后感染病毒,再加入苦参碱。倒置显微镜下观察各组心肌细胞病变效应(CPE)变化,流式细胞仪检测心肌细胞凋亡情况,免疫细胞化学法及Western blot法检测心肌细胞内磷酸化AKTSer-473蛋白的表达。结果与正常对照组比较,病毒组、苦参碱治疗组心肌细胞CPE变化明显,细胞凋亡率增加,磷酸化AKTSer-473蛋白表达上调(Pa<0.05)。与病毒组比较,苦参碱治疗组心肌细胞CPE变化不明显,细胞凋亡率降低,磷酸化AKTSer-473蛋白表达明显增强(Pa<0.05);病毒+Wort组心肌细胞CPE变化明显,心肌细胞凋亡率增多(Pa<0.05),磷酸化AKTSer-473蛋白的表达下调(P<0.05)。与苦参碱治疗组比较,苦参碱治疗+Wort组CPE变化明显及细胞凋亡率增多,磷酸化AKTSer-473蛋白表达减少(Pa<0.05)。结论苦参碱对CVB3感染心肌细胞具有保护作用,这种保护作用至少是部分通过PI3K/AKT信号通路介导的。Objective To explore the protective effect of matrine on cultured cardiomyocytes infected with coxsackie virus B3（CVB3） and its relationship with phosphatidylinositol 3-kinase/protein kinase B（PI3K/AKT） singnal pathway.Methods Newborn Wistar rats were separated by trypsin digestion method.The primary cardiomyocytes were collected and incubated with CVB3 to construct viral myocarditis model in vitro.Cardiomyocytes were randomly alloted to 5 groups： normal control group,not infected with CVB3,not added matrine and inhibitor of PI3K（wortmannin,Wort）;virus control group,only infected with CVB3,not added matrine and inhibitor;virus＋ Wort group,added inhibitor（200 nmol·L-1）,infected with CVB3;matrine treatment group,infected with CVB3,added matrine（0.3 g·L^-1）;matrine treatment＋Wort group,added inhibitor,infected with CVB3,added Matrine.In every group,cytopathic effects（CPE） were observed,cell apoptotic rate was detected by Annexin V/propidium iodide through flow cytometer.And the expression of Phospho-AKTSer-473 were detected by immunocytochemistry and Western blot,respectively.Results Compared with normal control group,CPE,the cardiomyocytes apoptotic rate and Phospho-AKTSer-473 expression were significantly increased in virus control group and matrine treatment group（Pa〈0.05）.Compared with virus control group,CPE was significantly reduced and the cardiomyocytes apoptotic rate was pretty low in matrine treatment group,matrine also increased the expression of Phospho-AKTSer-473（Pa〈0.05）.Wort increased CPE and the cardiomyocytes apoptotic rate,the expression of Phospho-AKTSer-473 was significantly lower in virus control＋Wort group（Pa〈0.05）.CPE and the cardiomyocytes apoptotic rate were significantly increased,the expression of Phospho-AKTSer-473 was lower in matrine treatment ＋Wort group as compared with matrine treatment group（Pa〈0.05）.Conclusions Matrine provides concrete protection against injury of cardiomyocytes infected with CVB3 which manifest decrease

关 键 词：苦参碱 柯萨奇B3型病毒 心肌细胞 磷脂酰肌醇3-激酶/蛋白激酶B通路 

分 类 号：R285.5[医药卫生—中药学]