法舒地尔对急性柯萨奇B3病毒性心肌炎小鼠的保护作用

The protective effect of fasudil on mice with coxsackievirus B3-induced acute viral myocarditis

目的探讨法舒地尔对柯萨奇病毒B3 (CVB3)病毒性心肌炎小鼠的作用及可能的机制。方法 90只雄性BALB/c小鼠分为病毒性心肌炎法舒地尔治疗组、病毒性心肌炎组和正常对照组。心肌炎模型建立后次日开始,法舒地尔治疗组每日腹腔注射法舒地尔(10mg/kg),其他两组注射等量0.9%氯化钠溶液,分别于第7天和第14天从各组抽取8只小鼠行超声心动图检查,随后取血和心脏标本,行HE染色观察心肌病理改变,实时荧光定量聚合酶链反应法检测心肌组织CVB3、Rho激酶2(ROCK2)和白细胞介素-17(IL-17)的m RNA表达,蛋白质印迹法检测心肌组织ROCK2、IL-17蛋白水平表达,酶联免疫吸附法检测血清炎症因子IL-17水平。结果与正常对照组比较,病毒性心肌炎组小鼠心功能恶化,心肌组织ROCK2、IL-17m RNA和蛋白水平及血清IL-17表达均升高,法舒地尔干预后显著降低小鼠心肌ROCK2水平,抑制CVB3复制和IL-17的表达,改善心功能。结论 CVB3病毒感染后小鼠心肌组织Rho激酶表达增高,法舒地尔通过抑制Rho激酶途径减少CVB3复制和IL-17表达,对病毒性心肌炎小鼠的心肌损害起到保护作用。

Objective To investigate the protective effect of fasudil on mice with acute viral myocarditis （VMC）induced by coxsackievirus B3 （CVB3） and its mechanisms. Methods 90 male BALB/c mice were randomly divided intofasudil group（myocarditis group treated with fasudil）, placebo group（myocarditis treated with placebo） and control group.One day later after myocardial model established, fasudil （10 mg ·kg^-1） was injected intraperitoneally daily in fasudil group,while same dose of 0.9% saline was injected intraperitoneally in control group and placebo group. 8 mice in each groupwere selected randomly on day7 or 14 for echocardiography examination and then the blood and hearts were obtained.Myocardial histopathology was evaluated with HE stain. mRNA expressions of CVB3, Rho kinase（ROCK2） and IL-17 weremeasured by PCR. The protein levels of myocardial ROCK2 and IL-17 were measured by western blotting. The serumIL-17 was measured by ELISA. Results The mRNA expressions and protein levels of ROCK2 and IL-17 expression,and serum IL-17 were increased and cardiac function decreased significantly in placebo group compared to controlgroup. Fasudil treatment significantly reduced myocardial ROCK2 level, inhibited CVB3 replication and IL-17 expression,and improved cardiac function. Conclusion The expression of Rho kinase in myocardium increases after CVB3infection. Fasudil protects mice against myocardial damage by inhibiting Rho kinase pathway and decreasing CVB3replication as well as IL-17 expression.