Expression And Mechanism Of Nrf2 And GPX3 In Vaginal Wall Tissue Of Patients With Pelvic Organ Prolapse

Purpose: By measuring Nrf2,GPX3 and collagen metabolism related protein in Pelvic organ prolapse(POP)group and control group in the vaginal wall tissue expression situation,analysis of Nrf2,GPX3 and correlation between collagen,preliminarily Nrf2 signaling pathway in patients with Pelvic organ prolapse vaginal wall mechanism in the organization,and lay a foundation for studying the pathogenesis of POP.Method: In this study,100 patients who underwent surgical treatment in shengjing hospital affiliated to China medical university from March 2018 to September 2018 were selected,including 50 patients of POP(case group)and 50 patients of total hysterectomy for benign uterine lesions(control group).The stages of all POP patients were Ⅲ to Ⅳ degrees.Collection of two groups of patients with vaginal wall tissue,using immunohistochemical method(SP),Western blot and rt-pcr method to detect the vaginal wall tissue of patients with two groups of Nrf2,GPX3,MMP-2 and TIMP-1,COLⅠ and COLⅢ expression,and analyze the Nrf2 and GPX3,COLⅠand COLⅢ whether there is a correlation between the expression of situation.Selecting the POP group and control group in all 3 cases of vaginal wall tissue fibroblasts primitive culture,using Western blot method and RT-PCR method to detect Nrf2,GPX3,MMP-2 and TIMP-1,COLⅠand COLⅢ expression.MTT assay was used to detect the proliferation of the two component fibroblasts.Nrf2 knock down model was constructed and divided into three groups: Blank control group(Blank group),empty vector group(NC group)and siRNA Nrf2 group.Using the method of RT-PCR and Western Blot technique to detect the above model of Nrf2,GPX3,MMP-2 and TIMP-1,COLⅠ and COLⅢ mRNA and protein expression level.MTT method was used to detect the cell proliferation in Blank control group(Blank group),empty vector group(NC group)and siRNA Nrf2 group.Results: Comparison of general conditions: the age of POP group and control group was 51.62±7.31 years old and 49.34±4.54 years old,respectively,with no significant difference between the two groups(P>0.05).The frequency of vaginal delivery was 2.82±1.24 times in the POP group and 2.50±0.94 times in the control group(P>0.05).BMI of POP group and control group was 25.51±3.57 and 25.51±3.57,respectively,with no significant difference between the two groups(P>0.05).Immunohistochemical results showed that Nrf2,GPX3,TIMP-1,COLⅠ and COLⅢ in expression rate in the POP group was lower than those of control group,difference was statistically significant(χ~2 = 11.66,χ~2 = 8.45,χ~2= 16.79,χ~2 = 9.55,χ~2 = 11.82,P < 0.05);The expression level of mmp-2 in the POP group was higher than that in the control group,and the difference was statistically significant(χ~2=14.64,P<0.05).Spearman correlation analysis showed that the expression of Nrf2 and GPX3 positively correlated(r = 0.61,P < 0.05),POP group COLⅠ and COL Ⅲ expression level is lower than the control group,and was positively correlated with the expression of Nrf2(r = 0.53,r = 0.42,P < 0.05);Western Blot method tests showed the POP group Nrf2,GPX3,TIMP-1,COLⅠ and COLⅢ protein expression level was lower than those of control group(t = 4.08,t = 2.98,t = 4.70,t = 10.54,t = 5.05,P < 0.05),the protein expression of MMP-2 levels higher in the POP group,difference was statistically significant(t = 9.22,P < 0.05).RT-PCR method tests showed the POP group Nrf2,GPX3,TIMP-1,COLⅠ and COLⅢ mRNA expression level was lower than those of control group(t = 7.22,t = 5.21,t = 7.60,t = 14.53,t = 5.07,P < 0.05),while the MMP-2 mRNA expression level is higher than the control group,difference was statistically significant(t = 12.87,P < 0.05).MTT method was used to detect the proliferation activity of the two component fiber cells,and the results showed that the proliferation capacity of POP component fiber cells was significantly lower than that of the control group,and the difference was statistically significant(P<0.05).Method of RT-PCR and Western blot method to detect siRNA Nrf2 transfection of groups of related indicators after vaginal wall fibroblasts mRNA and protein expression level of influence,the result shows: the Nrf2,GPX3,TIMP-1,COLⅠ and COLⅢ mRNA and protein expression level lowered(P < 0.05),the expression of MMP-2 levels increase(P < 0.05).MTT method was used to detect the proliferation activity of the vaginal fibroblasts after transfection with siRNA Nrf2,and the results showed that the proliferation capacity of the Nrf2 group of siRNA significantly decreased compared with the Blank group and the NC group(P<0.05).Conclusions: The loss of Nrf2 may promote the catabolism of collagen,leading to the occurrence of POP.Nrf2 deletion may be involved in the occurrence and development of POP by affecting the cell’s proliferation ability.