动物造模 z-bio 2022-06-08 307

　　Objective: To detect and analyze the expression of HBV-related indicators in the urine of HBV transgenic mice, and to further understand the biological characteristics of transgenic mice; and to determine the tissue source of HBV-related indicators in urine through various experimental methods.

　　Methods: Enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative PCR (real-time PCR) were used to detect HBV-related indicators in the urine of transgenic mice; and the expression of HBV transgene was inhibited by hydrodynamic transfection of mice and RNA interference methods. The source of its replication and expression tissue was determined by various experimental methods, such as the mouse experiment and the infection of normal mice with HBV positive serum from hepatitis B patients.

　　Results: HBsAg, HBeAg, and HBV-DNA were expressed in the urine of HBV transgenic mice, and the expression level of HBsAg was higher (6674±619.8 IU/mL), but lower than the expression level of HBsAg in serum (16470±2704 IU). /mL), the expression level of urinary HBsAg in male mice is higher than that in female mice; while the expression level of HBeAg in urine is lower, and the positive rate of HBeAg in urine is higher than that in blood, the expression level of HBeAg varies between individuals and between genders Significant difference; HBV-DNA content in urine reached 103~105copy/mL; no related antibody expression was detected. Experiments such as hydrodynamic transfection showed that the HBV-related indicators in the urine of transgenic mice originated from the replication and expression of the kidney, rather than being secreted by the liver into the blood circulation and excreted in the urine. The kidney is an independent expression site of HBV.

　　CONCLUSION: There are HBV-related indexes in the urine of transgenic mice, and they are expressed for a long time with the age of the month. The expression levels of HBsAg and HBV-DNA are relatively high and stable. The titer of HBsAg in male mice is higher than that of female mice. ;The expression level of HBeAg in male mice is higher and more stable than that in female mice; there is no expression of various antibodies in urine; the kidney is also an independent replication and expression site of HBV.