柯萨奇病毒A组16型与小鼠病毒性心肌炎的关联性及其药物干预研究

Relevance between Coxsackievirus A16 and viral myocarditis,and drug intervention

目的研究柯萨奇病毒A组16型(CVA16)与小鼠病毒性心肌炎的关联性,以及利巴韦林药物干预后的结果。方法将获取的临床标本分离后在非洲绿猴肾细胞中进行大量培养,选用致病性强的一株进行纯化、鉴定,培养出有一定毒力的致病株,以腹腔注射的方式感染刚出生的小鼠,小鼠共分为5组,每组10只,其中A组为正常对照组,B组为模型对照组,C组为低剂量的利巴韦林干预组,D组为中剂量的利巴韦林干预组,E组为高剂量的利巴韦林干预组,记录小鼠每日体质量、四肢活动情况、死亡情况,分别于感染后第3、7、11、15天获取小鼠的血清及心脏组织,采用酶联免疫吸附法(ELASA)试剂盒检测血清中的白细胞介素-6(IL-6),将心脏组织一部分用于病理切片,一部分用于提取RNA,进行反转录PCR(RT-PCR)实验。结果 B组小鼠的血清IL-6值明显升高,与A组比较,差异有统计学意义,并且RT-PCR的结果表明病毒在感染后的小鼠心肌细胞中存在,同时病理切片中可见明显的心肌炎性病变,利巴韦林药物干预之后,D组小鼠的体质量、病理切片炎性病变好转。结论 CVA16与病毒性心肌炎的发生有一定关联性,一方面病原本身对心肌有损伤,另一方面免疫内环境也参与其中,利巴韦林在治疗CVA16有关的心肌炎中有一定的临床意义。

Objective To study the relevance between Coxsackievirus A16 and viral myocarditis,and the results after ribavirin interven-tion. Methods We picked up the Coxsackievirus A16 from clinical specimens, cultured the vims in Vero cell, selected the one with strong pathogenicity,identified it again,and used the vims whose virulence was known to infect newborn mouse via intraperitoneal injec-tion. The mice were assigned into 5 groups with 10 in each. Group A was the normal control group,group B was the model group,and groups C,D,E were intervention groups with low,medium and high-dose ribavirin,respectively. We recorded the daily body mass,activi-ties of the limbs, and death of the mice. The serum and myocardium of the mice were obtained on the third /seven/eleven/fifteen day af-ter infection, the serum used for testing IL-6 by ELASA and part of the myocardium was used for pathological section examination, while the other part was used for extracting the RNA,tested by RT-PCR. Results Serum IL-6 levels in group B were significantly increased, and all showed certain trends in change,which were significantly different compared with group A. RT-PCR results proved that Coxsack-ievirus A16 expressed in myocardial cells of the infected mice,and pathological section indicated obvious myocardial inflammatory chan-ges. After ribavirin intervention, body mass and myocardial inflammatory changes were getting better in group D than in other groups. Conclusions Coxsackievirus A16 has some relevance with viral myocarditis. Both the pathogen itself and the immune internal environ-ment damage myocardium. Ribavirin shows clinical significance in the treatment of myocarditis with Coxsackievirus A16.