Study On Dynamic Changes Of Serum USP7 In Patients With Hepatitis B And Its Regulatory Effect On Apoptosis Factors

Objectives By detecting changes in the expression of Ubiquitin Specific Peptidase 7(USP7)and apoptosis factors in the serum of patients with hepatitis B.The deubiquitination of tumor protein p53(p53)by USP7 is verified in the hepatitis B cell model to investigate the mechanism of USP7 in hepatitis B,aiming to provide a theoretical basis for clinically more effective treatment of hepatitis B.Methods It is mainly divided into two parts: population and cell experiment.1 Population studies: According to the inclusion and exclusion criteria for hepatitis B,85 inpatients with hepatitis B in the Fifth Hospital of Shijiazhuang City were selected as the study subjects.The m RNA expression levels of USP7,p53,Bcl2 associated X protein(Bax)and B-cell lymphoma-2(Bcl-2)were detected by Real-time fluorescence quantitative PCR(RT-q PCR),and the protein expression levels of USP7,p53,Bax and Bcl2 were detected by enzymelinked immunosorbent assay(ELISA).The data of the population and the expression changes of USP7,p53,Bax and Bcl2 during the treatment were analyzed by repeated measure analysis of variance.The correlation between USP7 and alanine aminotransferase(ALT),aspartate aminotransferase(AST),Hepatitis B virus-hepadnavirus(HBV DNA),Hepatitis B surface antigen(HBs Ag),Hepatitis B e antigen(HBe Ag),p53,Bax,Bcl2 was analyzed by Pearson correlation.2 Cell experiment: The cells were divided into control group,si RNA-USP7 group,si RNA-NC group,ETV group,ETV+si RNA-USP7 group and ETV+si RNA-NC group on the basis of determining the optimal concentration of anti-HBV Entecavir(ETV).The m RNA expression levels of USP7,p53,Bax and Bcl2 were detected by PCR.The expression levels of ALT and AST were detected by Lai’s method.the HBVDNA load was detected.And the protein expressions of HBe Ag,HBs Ag and Bax and Bcl2 were detected by ELISA.Western Blot was used to detect the expression of USP7 and p53 proteins at different time points in each group,and immunocoprecipitate method was used to detect the interaction of USP7 and p53 proteins.single factor analysis of variance was used to compare whether the expression of cells in each group at different time points was statistically significant.P < 0.05 was considered to be statistically significant.Results 1 Population studies: With the increase of treatment time,the expression of USP7 in serum of hepatitis B patients decreased gradually(P < 0.05).HBV-DNA load,HBs Ag,HBe Ag,liver function indexes(ALT,AST),apoptosis regulator p53,pro-apoptotic factor Bax,and anti-apoptotic factor Bcl2 expression were also decreased with the prolongation of treatment time(P < 0.05),and the Bax/Bcl2 ratio was > 1.USP7 was positively correlated with ALT,AST,HBV-DNA,HBs Ag,HBe Ag,p53,Bcl2 and Bax(r = 0.522,0.509,0.478,0.549,0.556,0.716,0.676,0.566,all P < 0.05).2 Cell assay: The concentration was determined to be 25 μg/m L.USP7 in ETV group was gradually down-regulated with the increase of dosed time(P < 0.05),and the expression of USP7 at 24 h,48 h and 72 h was lower than that in control group.The changes of HBV-DNA load and HBs Ag,HBe Ag,ALT,AST,p53,Bax and Bcl2 expression levels were the same as USP7.USP7(si RNA-USP7group)was inhibited,and USP7 expression decreased with the extension of cell culture time(P < 0.05),and USP7 expression was lower than that of control group at all time points.HBV-DNA load and the expression levels of HBs Ag,HBe Ag,ALT,AST,p53,Bax and Bcl2 decreased after USP7 inhibition,and the change trend was the same as that of USP7 inhibition.At the same time,USP7(ETV+si RNA-USP7 group)was added and inhibited,and the expression of USP7 was lower than that of ETV group and si RNA-USP7 group at each time point,and the expression of USP7 was gradually down-regulated with the extension of culture time(P < 0.05).HBV-DNA load and HBs Ag,HBe Ag,ALT,AST,p53,Bax,Bcl2 also had the same expression changes.The Bax/Bcl2 ratio of each group at different time points was > 1.The verification results of the interaction between USP7 and p53 protein showed that there was indeed interaction between USP7 and p53 protein.Conclusions 1 USP7,p53,Bax and Bcl2 are decreased during the treatment of hepatitis B.2 USP7 is positively correlated with liver function indexes(AST,ALT),HBV-DNA,HBs Ag,HBe Ag,p53,Bcl2 and Bax.3 USP7 inhibits p53 ubiquitination by playing a role in ubiquitination.USP7 down-regulates p53 expression and reduces the expression of apoptotic factors Bax and Bcl2.Figure 22;Table 12;Reference 128...