动物造模,乙肝肝纤维化小鼠复合模型 z-bio 2022-11-03 264

　　Objective: To compare the virological and biochemical pathological characteristics of 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 liver fibrosis

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

　　Results: Except WT group and CCl4 group, serum HBsAg and HBeAg were positive in other groups by ELISA, and HBV DNA load was higher than 1.0 by fluorescent quantitative PCR × 104IU/mL. The content of HBsAg and HBeAg in the Tg+CCl4 group was higher than that in the rAAV+CCl4 group; The expression of HBcAg in rAAV group was significantly higher than that in Tg group. Compared with rAAV+CCl4, there was no significant difference in the results of HBsAg in Tg+CCl4 and rAAV+CCl4. The positive expression of HBcAg in rAAV+CCl4 group was significantly higher. The biochemical results showed that the contents of ALT, AST in serum and Hyp in liver tissue in CCl4 group, Tg+CCl4 group and rAAV+CCl4 group were significantly higher; Among them, the Hyp content in the Tg+CCl4 group was higher than that in the rAAV+CCl4 group. There was no significant difference between the AKP results and the WT group. The pathological results showed that the inflammation and collagen deposition were not obvious in the rAAV and Tg groups compared with the WT group; The inflammatory reaction and collagen deposition in CCl4 group, Tg+CCl4 group and rAAV+CCl4 group were significantly increased, and the collagen deposition in Tg+CCl4 group was higher than that in rAAV+CCl4 group

　　Conclusion: The two composite models meet the requirements of hepatitis B liver fibrosis model, and the differences mainly focus on virological indicators and pathological changes. rAAV+CCl4 group has advantages in HBV DNA load, and Tg+CCl4 group has more obvious fibrosis progress