苦参碱对病毒性心肌炎小鼠蛋白激酶B表达的影响

目的 评价苦参碱对柯萨奇B3型病毒（CVB3）感染的病毒性心肌炎（VMC）小鼠心肌组织蛋白激酶B（Akt）表达的影响,以探讨苦参碱保护心肌的作用机制.方法 实验采用雄性Balb/c小鼠连续3 d腹腔注射0.2ml 100TCID50的CVB3的方法制备VMC的动物模型.分为6组：苦参碱每日80 mg/kg、40 mg/kg、20 mg/kg治疗组,利巴韦林组、病毒组及正常对照组.药物于末次注射病毒的60 min后开始,连续腹腔注射10 d,正常组给予同体积的生理盐水.分别于给药后第5天和第10天处死小鼠,应用Tunel标记法检测各组心肌细胞凋亡情况;于给药后第10天应用免疫组化及Western Blot法检测心肌组织磷酸化AktSer-473蛋白的表达.结果 与正常组比较,病毒组心肌组织凋亡明显增加（P＜0.05）;与病毒组比较,苦参碱各剂量组心肌组织凋亡明显减少（P＜0.05）.于给药后第10天,苦参碱每日40 mg/kg组磷酸化AktSer-473蛋白的表达明显上调（P＜0.05）.结论 苦参碱可能通过促进磷酸化AktSer-473蛋白的表达,抑制CVB3感染的VMC小鼠心肌组织凋亡.

Effects of matrine on akt protein expression in mice with viral myocarditis

Objective To investigate the effects of Matrine on Akt protein expression in the myocardial tissue in mice with viral myocarditis(VMC) and to approach Matrine's protective mechanism.Methods Four-week-old healthy male Balb/c mice were inoculated intraperitoneally with 0.2ml of 100TCID50 Coxsackie virus B3(CVB3) every day(the.total is for 3 days) to construct VMC animal model. A total of 120 mice were randomly divided into 6 groups:Matrine treatment groups including high does(80 mg/kg), midst does (40 mg/kg) , lower does (20 mg/kg); Ribavirin group; virus group and control group.The medicine treatment groups were injected intraperitoneally with equal volume solution starting from 60minutes after CVB3 were inoculated in the last time, the total days of administering medicine is for10 days.And control group were injected with 0.2ml of 0.9% sodium chloride instead of medicine. Six animals were killed in the 5th and the 10th day after administering medicine in each group for determination apoptosis (by Tunel method). Protein expression of Phospho-AktSer-473 were detected in the 10th day after treatment (by immunohistochemistry and Western Blot respectively). Results Compared with control group, apoptotic cells in virus group were increased (P＜0.05). The groups of Matrine treatment can significantly decrease apoptosis of myocardial tissue of VMC animals, and Matrine(40 mg/kg) can significantly increase phosphorylation of Aktser-473 in the myocardial tissue as compared with virus group. Conclusion Matrine can reduce myocardium apoptosis of VMC animals by promoting phosphorylation of AktSer-473 in the myocardial tissue to provides concrete protection against CVB3 infection.