The Changes Of USP22 After Partial Hepatectomy In Mice

Objective: To explore the trend of ubiquitin specific peptidase 22(USP22)after partial hepatectomy in mice,and to explore its possible mechanism in liver regeneration,Which provides a theoretical basis for clinical research and development for the future。Method: In the experimental group,70% liver resection model was established by the classical Higgins and Anderson method.After centrifugation,the serum was taken from the inferior vena cava.The serum was analyzed by biochemical analysis.Alanine aminotransferase(ALT),aspartate Acid aminotransferase(AST)levels.After the mice were sacrificed,the regenerated liver weight of the residual liver was weighed and the liver regeneration rate was calculated.The expression of Ki67 was detected by immunohistochemistry.Ki67 positive expression was found in the nucleus of the nucleus,and 10 stained fields were selected at high magnification(40×).The results were determined by semi-quantitative scoring method.Protein extraction: RIPA lysate(containing protease inhibitor)to mention the total tissue protein,BCA method for protein quantification.Western blot analysis: SDS-PAGE electrophoresis to PVDF membrane,and finally in the darkroom film exposure development;results to USP22 / β-actin gray value of the target factor protein relative expression.RNA extraction: Total RNA was extracted using Trizol and reverse transcribed to obtain total c DNA.RT-PCR detection: the use of c DNA as a template for amplification,in accordance with the proportion of the reaction system(each sample repeated 3 times).The results were analyzed using SDS 1.1 software.Statistical analysis: Data analysis using SPSS18.0 statistical software,measurement data with mean ± standard deviation,the expression level of the target factor in different groups using t test.Results: 1.The rate of liver regeneration after PH in mice was mainly after24 h,the final mouse liver regeneration rate can be stable more than 70%.The changes of liver function transaminase in mice were significantly higher than those in control group(P <0.05).The changes of ALT and AST were significantly increased in 6 h-24 h,and then decreased rapidly until the control group.2.The results of immunohistochemical analysis showed that the number of Ki-67 positive cells increased at 12 h after operation and reached the peak at 48 h and 96 h after operation,and the number of positive cells decreased gradually(p <0.01).3.Western blotting of USP22 protein showed that the expression level of USP22 protein was decreased sharply after operation,and then the protein expression level increased gradually with the prolongation of postoperative time,the difference was statistically significant(p <0.01).4.The m RNA expression of USP22,β-Catenin and Cyclin D1 in regenerated liver: The RT-PCR results of mouse liver at different time points after resection showed that the expression level of Cyclin D1 m RNA was higher than that of control group at 48 h,And then gradually decreased to the normal level.The expression of β-Catenin was also decreased after operation,and the expression level of β-Catenin was increased after 24 hours,and reached the peak at 96 h.The expression of USP22 was similar to that of 48 h,and the difference was statistically significant Meaning(p <0.01).Conclusion: The expression of USP22 protein decreased first and then increased.And USP22 and β-Catenin m RNA expression is closely related,indicating that in the process of liver regeneration,USP22 and Wnt / β-catenin pathway are closely linked.