动物造模,乙肝肝纤维化 z-bio 2023-05-25 489

　　Objective: To use C57BL/6N-Tg(1.28HBV)/Vst hepatitis B virus transgenic mice and rAAV8-1.3HBV adeno-associated virus-transfected mice combined with CCl4 intraperitoneal injection to induce hepatitis B fibrosis mouse model, and compare the two mice Virological and biochemical pathological characteristics of the model.

　　Methods: The experiment was divided into wild-type control group (WT), rAAV8-1.3HBV transfection group (rAAV), CCl4 group (CCl4), rAAV8-1.3HBV combined CCl4 model group (rAAV+CCl4), C57BL/6N-Tg (1.28 HBV)/Vst hepatitis B virus transgenic group (Tg) and transgenic compound CCl4 group (Tg+CCl4), the model was established for a total of 12 weeks. The serum HBsAg and HBeAg levels were determined by ELISA kit, and the serum HBV-DNA load was detected by fluorescence quantitative PCR. The expression of HBsAg and HBcAg in the liver was observed by immunohistochemical staining of paraffin sections of liver tissue, serum ALT, AST and AKP were determined by biochemical kits, and the content of hydroxyproline (Hyp) in liver tissue was detected by hydrochloric acid hydrolysis method. HE staining and Sirius red staining were used to observe Pathological changes such as inflammation and collagen.

　　Results: Except for the WT group and the CCl4 group, the serum HBsAg and HBeAg detected by ELISA in the other groups were all positive, and the HBV-DNA load detected by fluorescence quantitative PCR was higher than 1.0×104 IU/mL. Among them, the Tg+CCl4 group HBsAg and HBeAg content The level of HBV-DNA in the rAAV, rAAV+CCl4 group was higher than that in the Tg, Tg+CCl4 group. Immunohistochemistry showed that except for the WT group and the CCl4 group, the HBsAg and HBcAg in the liver tissues of the other groups were all positive Compared with Tg group, HBsAg expression in rAAV group decreased; compared with Tg group, HBcAg expression in rAAV group increased significantly. Compared with rAAV+CCl4, Tg+CCl4 showed no significant difference in HBsAg expression, but rAAV+CCl4 group had significantly higher HBcAg expression. Biochemical The results showed that: CCl4 group, Tg+CCl4 group and rAAV+CCl4 group were significantly increased in serum ALT, AST and Hyp content in liver tissue; among them, the Hyp content in Tg+CCl4 group was higher than that in rAAV+CCl4 group. Compared with WT group, AKP results showed no significant difference. The pathological results showed that compared with the WT group, the inflammation and collagen deposition in the rAAV and Tg groups were not obvious; in the CCl4 group, the Tg+CCl4 group, and the rAAV+CCl4 group, the inflammation and collagen deposition were significantly increased, and the collagen deposition in the Tg+CCl4 group was higher than that in the Tg+CCl4 group. rAAV+CCl4 group.

　　Conclusion: The two composite models meet the requirements of hepatitis B fibrosis model, and the differences are mainly concentrated in virological indicators and pathological changes. The rAAV+CCl4 group has an advantage in HBV-DNA load, and the Tg+CCl4 group has a more advanced fibrosis progression. obvious.