动物造模,肝纤维化小鼠复合模型 z-bio 2021-10-27 475

　　Objective: Combine C57BL/6N-Tg (1.28HBV)/Vst hepatitis B virus transgenic mice and rAAV8-1.3HBV adeno-associated virus transfected mice with intraperitoneal injection of CCl4 to produce type B mice that can induce liver fibrosis And compare the viral and biochemical pathological characteristics of the two mouse models.

　　 Method: The experiment was divided into wild control group (WT), rAAV8-1.3HBV transfection group (rAAV), CCl4 group (CCl4), rAAV8-1.3HBV combined with CCl4 model group (rAAV). + CCl4), C57BL/6N-Tg (1.28HBV)/Vst hepatitis B virus transgenic group (Tg), transgenic compound CCl4 group (Tg + CCl4), a total of 12 weeks of modeling? ELISA kit, used to detect serum HBsAg and HBeAg levels, fluorescence quantitative PCR to detect serum HBV-DNA load, and immunohistochemical staining of liver tissue paraffin sections of HBsAg and HBcAg to observe the expression and serum ALT? AST? AKP was measured with a biochemical kit, and hydroxyproline in liver tissue was detected with hydrochloric acid (Hyp) content. HE staining and Silius red staining can be used to observe pathological changes, such as inflammation and collagen?

　　Result: Except for the WT and CCl4 groups, the ELISA tests of serum HBsAg and HBeAg were all positive, and did the fluorescence quantitative PCR detection volume of HBV-DNA exceed 1.0×104 IU/mL? HBsAg in the Tg + CCl4 group? Is the HBeAg content higher than the rAAV + CCl4 group? AAV? Is the HBV-DNA level of Tg in the AAV + CCl4 group Tg? Is it higher than the Tg + CCl4 group? Immunohistochemistry showed: Except for WT and CCl4 groups, are the liver tissues of other groups positive for HBsAg and HBcAg? Compared with the Tg group, the expression of HBsAg in the rAAV group was reduced; compared with the Tg group, did the expression of HBcAg in the rAAV group increase significantly? Compared with the results of Tg + CCl4 and rAAV + CCl4 HBsAg, there is no significant difference. The positive expression of HBcAg in the AAV + CCl4 group increased significantly. The biochemical results showed that in the CCl4 group, Tg + CCl4 group, rAAV + CCl4 group, serum ALT, AST, and Hyp content in liver tissue increased significantly. Is the Hyp content of the Tg + CCl4 group higher than that of the rAAV + CCl4 group? Is there a significant difference between the AKP results and the WT group? The pathological results showed that the inflammation and collagen deposition in rAAV and Tg groups were not obvious compared with WT group and CCl4 group. Tg + CCl4 group? The inflammation and collagen deposition in the AAV + CCl4 group increased significantly. Among them, is the collagen deposition of the Tg + CCl4 group higher than that of the rAAV + CCl4 group?

　　 Conclusion: These two composite models meet the requirements of hepatitis B fibrosis model. The main difference is the viral indicators and pathological changes? Does the AAV + CCl4 group benefit from the HBV-DNA load, and does the Tg + CCl4 group have more obvious fibrosis progression?