肠道病毒71型ICR乳鼠感染模型的建立

Establishment of enterovirus 71-infected ICR neonatal mice model

目的:建立肠道病毒71型(Enterovirus 71,EV71)感染ICR乳鼠模型,为该病毒疫苗和抗病毒药物的研究提供实用的动物模型。方法:将本课题组自行分离鉴定的EV71 G03株经颅内接种感染1日龄SPF级ICR小鼠,定期对濒死的小鼠实施安乐死,采集心肌、肺组织、骨骼肌、小肠和脑组织,RT-PCR检测病毒RNA、HE染色观察病理形态学、免疫组织化学分析及蛋白质印迹鉴定病毒VP1蛋白表达。结果:经颅内途径感染ICR乳鼠后,小鼠出现消瘦、精神麻痹、弓背、肢体麻痹瘫痪、死亡等症状;病理学观察发现,在感染后小鼠的心肌、肺组织、骨骼肌、小肠和脑组织等出现特定的病理变化,如出血、炎症细胞浸润等。免疫组化显示感染后所取小鼠的5种组织都有EV71病毒特异性分布。蛋白质印迹分析显示EV71 VP1蛋白在所取小鼠5种组织均有表达。结论:成功建立了EV71感染1日龄ICR乳鼠模型。

Objective ： To establish an EV71 infection model in ICR neonatal mice and provide a practical animal model for evaluation of EV71 vaccines and antiviral drugs. Methods： The 1-day ICR mice were infected with the EV71 G03 virus strain through intracerebroventricular injection to establish animal models. After routine euthanizingly killed animals, their various organs and tissues were collected to perform RT-PCR analysis of the viral nucleotide acids. In addition, pathological observation, immuno- histochemical staining and the Western blotting were used to identify the expression of viral protein 1 （VP1） . Results ： Pathological observation found that after infection, specific pathological changes, such as bleeding, inflammatory cell infiltration, appeared in the myocardium tissue, lung tissue, skeletal muscle ,small intestine, brain tissue and other organs of mice. Immunohistochemical staining showed that all five tissues of the infected mice had EV71 virus-specific antigen distribution. Western blotting analysis demonstrated that EV71 VP1 protein was expressed in all five tissues. Conclusion The one-day-old ICR neonatal EV71-infected mouse model was successfully established.