柯萨奇病毒B3诱导病毒性心肌炎小鼠心肌细胞焦亡的实验研究

Experiment Research of Coxsachie Virus B3 Induced Myocardial Pyrolysis in Mice with Viral Myocarditis

目的探讨柯萨奇病毒B3(CVB3)是否可通过诱导心肌发生细胞焦亡促进心肌炎的进展。方法采用CVB3感染Bal b/c小鼠构建病毒性心肌炎动物模型,分为对照组(10只)和实验组(20只)。对照组腹腔注射0.1 mL无菌生理盐水,实验组每只小鼠腹腔注射0.1 mL含10^(5)半数组织培养感染剂量(TCID_(50))CVB3病毒液,分别于感染后第3天、第7天随机选取10只小鼠,摘眼球收集全血,处死后摘取心脏。用酶联免疫吸附试验(ELISA)检测小鼠血清中白细胞介素-1β(IL-1β)水平。采用实时定量聚合酶链式反应(RT-qPCR)检测NLRP3炎性小体及半胱天冬氨酸蛋白酶-1(Caspase-1)mRNA水平。采用蛋白质印迹法(Western Blot)检测Caspase-1、半胱天冬氨酸蛋白酶酶原(pro-Caspase-1)蛋白表达水平。结果实验组第3天、第7天小鼠心肌组织中NLRP3炎性小体及Caspase-1 mRNA表达较对照组明显增加(P<0.05);pro-Caspase-1、Caspase-1的蛋白表达水平较对照组明显增加(P<0.05);血清IL-1β的表达较对照组明显增加(P<0.05)。结论CVB3可通过诱导心肌发生细胞焦亡促进心肌炎的进展。

Objective To investigate whether Coxsachie virus B3(CVB3)can promote the progression of myocarditis by inducing myocardial pyrolysis.Methods Balb/c mice infected with CVB3 were used to construct viral myocarditis models.The mice were divided into control group(n=10)and experimental group(n=20).The mice in control group were intraperitoneally injected with 0.1 mL sterile normal saline,and each mouse in experimental group was intraperitoneally injected with 10^(5) TCID_(50) CVB3 virus solution,respectively.At the 3rd day and 7th day,10 mice were randomly selected,the eyeballs were removed to collect the whole blood,and the hearts were removed after execution.The level of IL-1β in mouse serum was detected by enzyme linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the levels of inflammasome NLRP3 and Caspase-1 mRNA.Western Blot was used to detect the protein levels of Caspase-1 and pro-Caspase-1.Results The expression of NLRP3 and Caspase-1 mRNA in the myocardial tissue at 3rd and 7th day was significantly increased compared with that in control group(P<0.05).The protein expression levels of pro-Caspase-1 and Caspase-1 in the myocardial tissue at 3rd and 7th day in experimental group were higher than those of control group(P<0.05).The expression of IL-1βin the serum at 3rd and 7th day in experimental group was higher than that of control group(P<0.05).Conclusion CVB3 can promote the progression of myocarditis by inducing myocardial pyrolysis.