动物模型,HCV z-bo 2020-08-12 538

　　(1) Replication method The method is mainly to prepare transgenic mice by male pronucleus microinjection. In order to study the pathogenicity of HCV, scientists have studied many mice that have been partially transfected with HCV genes. The fragment containing HCV 5'UTR and structural gene (C1+E1+E2) was cut from the pHCS plasmid by restriction enzymes, and then cut with the intermediate vector pMT, and then inserted into the BamHI EcoRI site of human pcDNA3 (located under the CMV promoter) , The plasmid was named pCBE. Microinjection is used to inject the transgene into the promale pronucleus of the fertilized egg of the mouse, the injection amount is 1～2pl/egg, and then transplanted into the fallopian tube of a 6-week-old pseudo-pregnant female mouse. There are about 20 fallopian tubes on each side. Birth after development.

　　(2) Model characteristics The study found that in mice expressing the HCV non-structural protein Ns5A gene, no liver tissue lesions and direct cell pathogenicity were observed. The most significant manifestation in transgenic mice expressing HCV core protein is to induce liver fat changes and hepatocellular carcinogenesis, which is also a phenomenon that occurs in many chronic HCV infections. In mice transfected with the HCV core protein gene, lipid peroxidation and mitochondrial DNA damage caused by the increase of reactive oxygen species were also observed. It is inferred that the occurrence of hepatocellular carcinoma may be related to the increase of reactive oxygen species. Damage to DNA is related. At the same time, hepatic steatosis and hepatocellular carcinoma were also observed in a genetically modified gene (Core/E1/E2/P7) and a mouse strain transfected with complete HCV ORF.

　　(3) Comparative medicine Hepatitis C virus (HCV) is the main pathogen that causes hepatitis after blood transfusion. In addition to acute and chronic hepatitis, HCV infection can also cause liver cirrhosis and liver cancer. Due to the lack of a suitable tissue culture system and widely used animal models, it is impossible to study the effect of HCV on direct damage to liver cells. Therefore, the pathogenesis of HCV has not yet been fully understood. It may be caused by the direct action of the virus or the host's immune defense mechanism against the virus. The transgenic mouse system is considered to be the best system for studying the regulation of gene expression, and it can explore the regularity of gene activity and its phenotypic effect from the perspective of space and time. The establishment of the HCV structural gene mouse model is the establishment of a HCV gene transgenic mouse model and is used to study the mechanism of HCV infecting cells, the assembly and intracellular transport of HCV strains in liver cells, and the effect of HCV gene replication on liver cells The virulence, as well as the immunopathology of HCV structural proteins in the liver, etc., provide important technical guarantees.