HCV感染树鼩模型中腹部横切手术及肝门静脉注射新方法的建立

The establishment of new method for abdominal transverse operation and hepatic portal vein injection in the tree shrew model of HCV infection

目的采用腹部横切手术及肝门静脉注射法将丙型肝炎病毒(hepatitis c virus,HCV)迅速足量注射接种到树鼩体内肝脏,提高HCV对树鼩的感染率。方法采用横切打开树鼩腹腔,找到肝门静脉,进行肝门静脉注射接种HCV,手术后1周进行采血、ALT检测、HCV病毒载量检测、肝脏病理检测及HCV-RNA抗原检测。结果横切手术及肝门静脉注射20只树鼩,手术后经护理恢复,树鼩伤口恢复良好,手术成功率及肝门静脉注射准确率均达100%,保证HCV完全快速进入树鼩肝脏。有60%(12/20)树鼩出现一过性的病毒血症或间歇性病毒血症,感染树鼩的病毒载量最高可达到1x105拷贝/mL,肝功能中ALT指标最高可达到192μ/L,肝脏活体组织病理检查发现不同程度的肝炎症状,肝组织能检测到HCV-RNA抗原。结论腹部横切具有充分暴露肝门静脉、操作方便、符合解剖生理、组织创伤较小、伤口易于愈合等优点,适用于肝门静脉注射,同时提高了感染率,该技术方法为进一步深入研究HCV感染树鼩奠定基础,对于其它相关实验动物和动物实验具有一定借鉴经验和参考价值。

Objective HCV will be inoculated rapidly enough to tree shrew liver in vivo by using abdominal transverse operation and hepatic portal vein injection , to improve the rate of HCV infection in tree shrews. Methods Tree shrew peritoneal were opened by the transverse abdominal operation, the hepatic portal vein were found, HCV were inoculated by hepatic portal vein injection, after 1 week operation, blood, ALT detection, HCV viral load testing, liver pathology and HCV-RNA antigen detection. Results 20 tree shrews are transverse operation and hepatic portal vein injection, after the nursing care in the recovery operation, tree shrew wound recovered well, The success rate of operation and hepatic portal vein injection accuracy rate reached 100% , HCV is completely into the tree shrew liver. There are 60% （12/20） of tree shrew had viremia or intermittent viremia, viral load of infection tree shrew can reach 1×10^5 copies/ml, ALT index of liver function can reach 192μ/L, liver biopsy pathological examination showed different levels of symptoms of hepatitis, HCV-RNA antigen can be detected, in liver tissues. Conclusion Abdominal transverse has many advantages , including fully exposed the hepatic portal vein, convenient operation, in accordance with physiological solution planing, less tissue trauma,, good wound healing and so on, it is suitable for hepatic portal vein injection and increase the rate of infection, which lays a foundation for the further study of HCV infection of tree shrew, and has certain experience and reference value for other related experimental animal and animal experiment.