动物造模,药效评价,HCV感染模型 z-bio 2024-07-26 399

　　1. Modeling method

　　(1) Experimental animals: 4-6 week old clean grade male SD rats, weighing 113.2 ± 1.8g.

　　(2) Strain: Hepatitis C virus core region cDNA (HCV Core) was amplified by PCR, and recombinant adenovirus (rAAV) vector mediated virus recombination technology was used to prepare recombinant adenovirus containing HCV Core. The recombinant adenovirus has a potency of 2.41 × 10 ^ 11/ml.

　　Infection method: After anesthesia, 40% of the liver of 20 rats was removed, and 20 μ l of rAAV containing HCV Core gene was directly injected into the remaining liver lobes. Another 20 rats were taken and injected with empty vector rAAV as a control using the same surgical procedure. Routine postoperative feeding.

　　2. The modeling results showed that 10 rats were euthanized at 1 and 4 months after surgery, and liver tissues were taken for Southern blotting and Dot blotting hybridization to measure the integration of HCV Core genes and mRNA expression in the liver of the rats. The non site integration of HCV Core genes and Dot blot hybridization results of liver cell mRNA were all positive, indicating the expression of HCV Core mRNA.

　　Overall, an ideal animal model is a tool for studying human diseases, evaluating treatment strategies, and assessing drug safety. However, the low sensitivity of animals other than humans and chimpanzees to HCV has limited the establishment of animal models.

　　The transgenic mouse model expressing HCV protein has certain application value in studying the interaction between HCV and humans, as well as the liver pathology caused by HCV infection. However, it is not a true animal model and cannot be used for research on the life cycle of HCV, virus entry and replication, as well as the development of antiviral drugs and vaccines. The ideal mouse model should support HCV infection. Researchers have identified the cell surface receptor proteins that determine the host range and tissue tropism of the virus. The interaction between HCV and its receptors involves many steps, with the virus simultaneously or sequentially utilizing multiple receptors to bind to the cell surface and then enter the cell interior. Interestingly, although HCV cannot infect mouse liver cells, it can replicate in mouse liver cancer cell lines. The key to solving the narrow range of hosts susceptible to HCV infection in the future may depend on establishing transgenic mice expressing human HCV receptor proteins. CD81 is one of the receptors for HCV infection, however, transgenic mice expressing human CD81 protein in the liver are not sensitive to HCV, indicating that human CD81 protein is not the only determinant of HCV infection in mice. Therefore, constructing transgenic mice that allow HCV infection requires the simultaneous expression of several human HCV receptor proteins, such as SCARB1, CLDN1, and OCLN. In addition, establishing a transgenic mouse model capable of infecting HCV can help us better understand the immunopathological mechanisms involved in HCV infection.