| Objective:In this study,the expression and correlation of HOXA11 gene and key proteins Wnt16,β-catenin in Wnt/ β-catenin signal pathway and its downstream gene MMP2 in sacral ligament tissue of patients with pelvic organ prolapse were determined to explore the role of HOXA11 gene and Wnt/ β-catenin signal pathway in the pathogenesis of POP,so as to provide new targets and ideas for the prevention and treatment of POP.Methods:20 cases of sacral ligament tissue obtained from laparoscopic hysterectomy and vaginal sacral fixation due to pelvic organ prolapse(POP III-IV)in the Department of Gynecology of the second Hospital of Shanxi Medical University from October 2019 to October 2020 were selected as the POP group,and 20 cases of sacral ligament tissue who underwent transabdominal or laparoscopic total hysterectomy for uterine benign lesions in the same period were selected as the control group.The histological characteristics of the collected uterosacral ligament were observed by HE staining,the content,morphology and arrangement of collagen fibers in the sacral ligament were observed by Masson staining,the distribution and expression of HOXA11,Wnt16,β-catenin and MMP2 in the sacral ligament were detected by immunohistochemical method,and the expression of HOXA11,Wnt16,β-catenin and MMP2 in the sacral ligament was detected by Western-blot.In the POP group,the correlation between HOXA11 and each index was analyzed.Results:1.HE staining resultsA large number of fibrous connective tissue can be seen in the uterine sacral ligament of the control group,and the tissue is arranged densely and in a bundle shape.However,the number of fibrous connective tissue in the POP group was significantly reduced,and the structure was loose,disordered and disordered,in which the broken fibrous tissue was seen,and the number of fibroblasts was less than that in the control group.2.Masson staining resultsThe histopathological features were observed by Masson staining.The collagen fibers were blue-green and the muscle fibers were red.In the control group,collagen fibers were arranged neatly and continuously in wavy shape,while in the uterosacral ligaments of POP patients,the number of collagen fibers decreased significantly,arranged irregularly and broken in fragments,and the overall staining was lighter.3.Results of immunohistochemical detectionImmunohistochemical staining showed that the expression of HOXA11,Wnt16 andβ-catenin in the uterosacral ligament in the POP group was lower than that in the control group.but the expression level of MMP2 in the POP group was significantly higher than that in the control group.Semi-quantitative analysis of staining target protein was carried out by Image-Pro Plus 6.0software(IPP 6.0),the expression of HOXA11,Wnt16 andβ-catenin in uterine sacral ligament of POP group was 0.000931 ±0.000163,0.00220±0.00076 and 0.00109 ±0.000160 respectively,which was significantly lower than that of control group(0.00259 ±0.000931,0.00485 ±0.00130 and 0.00191 ±0.000439),The expression of MMP2 protein in the control group(0.00332 ±0.00122)was significantly higher than that in the control group(0.000719 ±0.000107),The difference was statistically significant(P< 0.05).4.Western-blot test resultsIn the uterosacral ligament tissue,the expression of HOXA11,Wnt16 and β-catenin in the POP group was 0.253 ±0.029,0.298 ±0.024 and 0.212 ±0.0377 respectively,which was significantly lower than that in the control group(0.319 ±0.029,0.371 ±0.045,0.425±0.0689),while the expression of MMP2 was 0.674 ±0.056 in the control group,which was significantly higher than that in the control group(0.290 ±0.054),The difference was statistically significant(P < 0.05).5.Analysis of correlation between HOXA11 and various testing IndexesSemi-quantitative immunohistochemical analysis showed that in POP group,the expression of HOXA11 was positively correlated with Wnt16 and β-catenin(r = 0.496,r=0.451),while the expression of HOXA11 was negatively correlated with that of MMP2(r=-0.686).Through the analysis of Western-blot results,it was further verified that HOXA11 was positively correlated with Wnt16 and β-catenin expression,and negatively correlated with MMP2 expression in POP group.The correlation coefficients were(r=0.490,r=0.524,r=-0.621).Conclusion:1.Collagen metabolism is closely related to the occurrence and development of POP.2.The expressions of HOXA11 and Wnt16 and β-catenin,the key proteins in Wnt/β-catenin signaling pathway,were decreased in the uterosacral ligament tissues of patients with pelvic organ prolapse,and the occurrence and development of POP may be related to the inhibition of their expression.3.The expression of HOXA11 was positively correlated with the expression of key protein in the Wnt/β-catenin signal pathway and negatively correlated with the expression of MMP2,which may be involved in the pathogenesis of POP. |
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