The Role Of Wnt/?-catenin Signaling Pathway And WNK1 In Cardinal Ligaments Of Female Patients With Pelvic Organ Prolapse

Pelvic organ prolapse(POP)is caused by pelvic floor support weak decrease in position and function of the pelvic cavity.It is a complex disease caused by multiple factors,multiple genes and multiple links.It is of great significance to understand the pathogenesis of POP to explore the role of the pop-susceptible genes and their involved signaling pathways in the occurrence and development of POP.Studies have shown that WNK1 gene mutation is involved in the development of POP,which is the pathogenic gene of POP.WNK1 also regulates the activation and shutdown of the Wnt/?-catenin signaling pathway.At the same time,it was found that the mechanism of POP was closely related to the abnormal Wnt/?-catenin signal pathway.However,the role of WNK1 gene and Wnt/?-catenin signaling pathway in the occurrence and development of POP and its correlation are not yet known.The stability of the status and function of the fibroblasts and collagen fibers in the pelvic floor supporting tissues can make the pelvic floor tissues function normally.The activation of fibroblasts is regulated by the Wnt/?-catenin signaling pathway.However,it is not known whether the Wnt/?-catenin signaling pathway affects the activity of fibroblasts and collagen expression in pelvic floor supporting tissues.ObjectiveTo analyze the expression of WNK1?Wnt16,?-catenin,FZD5 and COL1 in the main ligament of patients with pelvic organ prolapse.To explore the role and mechanism of WNK1 gene and Wnt/?-catenin signaling pathway in the pathogenesis of POP,and provide new diagnostic ideas and therapeutic approaches for POP from the molecular level.Materials and MethodsFrom January 2017 to May 2018,Twenty-seven patients with pelvic organ prolapse(POP)admitted to our hospital as POP group.According to the pelvic organ prolapse quantitative indexing method,all grade III-IV,the surgical approach is selected for transvaginal hysterectomy.Twenty-one patients with non-pelvic dysfunction of gynecological benign disease at the same time were selected as the control group.The operation was performed by laparoscopic,abdominal or transvaginal hysterectomy.The morphology and quantity of smooth muscle bundles and fibroblasts were observed by HE staining,and the number and morphology of collagen fibers were observed by Masson staining.The mRNA and protein expression levels of Wnt/?-catenin signaling pathway(Wnt16,?-catenin,FZD5),WNK1 gene and COL1 were detected by RT-qPCR and Western-blot.Result 1.HE staining and Masson stainingHE staining showed that compared with the control group,the smooth muscle bundles in the POP group were atrophied and broken,and the number of fibroblasts was significantly reduced and disordered.Masson staining showed that the collagen fibers in the POP group were sparse,finely divided and broken compared with the control group.2.RT-qPCRThe mRNA expressions of WNK1,Wnt16,?-catenin and COL1 in the main ligament of the control group were 1.115±0.078,1.116±0.053,4.316±0.286,1.066±0.072.The POP group was 0.868±0.061,0.884±0.048,3.451±0.214,and 0.661±0.008.The mRNA expression of the above factors in the POP group was significantly lower than that of the control group,and the difference was statistically significant(P=0.0145,P=0.0023,P=0.0171,P=0.0002)?The mRNA expression of FZD5 in the main ligament of the control group was 1.262±0.107.the POP group was 1.810±0.114.Its significantly higher than the control group,the difference was statistically significant(P=0.0013).3.Western-blotThe measured values of Wnt16,?-catenin,WNK1,and COL1 in the main ligament of the control group were 0.584±0.032,0.558±0.0378,0.740±0.0576,and 0.663±0.0362.The POP group was 0.427±0.028,0.347±0.029,and 0.529±0.0378,0.561±0.029.Compared with the corresponding measured values,the expression of POP group was significantly down-regulated,and the difference was statistically significant(P=0.0006,P<0.001,P=0.0027,P=0.0316).4.Association analysis between WNK1 and Wnt/?-catenin signaling pathwayIn the POP group,WNK1 was positively correlated with the expression of Wnt/?-catenin signaling pathways Wnt16 and ?-catenin(r=0.480,r=0.471),and negatively correlated with FZD5 expression(r=-0.512).5.High risk factor analysisThe expression of WNK1,Wnt16,?-catenin and COL1 in menopausal ? 5 years group was significantly lower than that in menopause time < 5 years,and the expression of FZD5 was increased,the difference was statistically significant(P=0.031,P=0.0356,P=0.0189,P=0.0269,P =0.0327).The number of vaginal delivery < 2 times group WNK1,?-catenin and COL1 was significantly lower than that of vaginal delivery ? 2 times,the difference was statistically significant(P=0.0207,P=0.0412,P=0.0118).ConclusionThe expressions of WNK1 and Wnt/?-catenin signaling pathways,Wnt16 and ?-catenin,were decreased in the ligament tissue of patients with pelvic organ prolapse.The development of POP may be related to the inhibition of expression.WNK1 gene is positively correlated with the expression of Wnt/?-catenin signaling pathway and may be involved in the pathogenesis of POP.