动物造模,HBV转基因 z-bio 2021-12-13 622

　　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 the transgenic mice; and to determine the tissue source of HBV-related indicators in the urine through a variety of experimental methods.

　　Methods: Enzyme-linked immunosorbent assay (ELISA) and real-time PCR (real-time PCR) were used to detect HBV-related indicators in the urine of transgenic mice; and hydrodynamically transfected mice experiments and RNA interference methods were used to inhibit HBV expression of transgenes Various experimental methods, such as the mouse experiment and the experiment of HBV positive sera from hepatitis B patients infecting normal mice, determine the source of its replication and expression tissues.

　　Results: There were HBsAg, HBeAg, and HBV-DNA expression in the urine of HBV transgenic mice. 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 HBsAg in male mice in urine 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. Significant difference; the content of HBV-DNA in urine reached 103~105copy/mL; no related antibody expression was detected. Experiments such as hydrodynamic transfection have shown that the HBV-related indicators in the urine of transgenic mice are derived from the replication and expression of the kidney, rather than secreted by the liver into the blood circulation and excreted in the urine. The kidney is an independent expression site for HBV.

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