WNT16B对体外培养宫骶韧带成纤维细胞增殖和胶原分泌的影响  

Effects of WNT16B on proliferation and collagen secretion of uterosacral ligament fibroblasts in vitro

作　　者：李蕾[1] 陈娟[1] 史宏晖[1] 朱兰[1] LI Lei;CHEN Juan;SHI Hong-hui;ZHU Lan(Department of Obstetrics&Gynecology,National Clinical Research Center for Obstetric&Gynecologic Diseases,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730)

机构地区：[1]中国医学科学院北京协和医学院北京协和医院普通妇科中心,国家妇产疾病临床医学研究中心,北京100730

出　　处：《生殖医学杂志》2022年第9期1267-1272,共6页Journal of Reproductive Medicine

基　　金：国家自然科学基金(81070474,82171621)。

摘　　要：目的研究WNT家族分泌型糖蛋白WNT16B对体外培养宫骶韧带成纤维细胞增殖和胶原蛋白分泌的影响,从分子水平解析盆腔器官脱垂(POP)的发病机制。方法选择2018年10月至2021年6月北京协和医院收治的4例POP患者,按POP定量(POP-Q)分度法均≥Ⅲ度;选择非POP的妇科良性疾病患者2例作为对照。取宫骶韧带组织标本进行韧带成纤维细胞的原代培养;构建WNT16B过表达载体并转染原代培养的成纤维细胞;RT-PCR法检测成纤维细胞WNT16B和I型胶原蛋白mRNA的表达水平;实时无标记细胞分析系统检测细胞增殖活性。结果POP患者宫骶韧带成纤维细胞WNT16B mRNA的表达水平显著低于非POP对照患者(P<0.001)。过表达WNT16B载体转染POP患者原代成纤维细胞后,细胞倍增时间显著缩短(P<0.05),I型胶原蛋白的mRNA表达水平显著提高(P<0.05)。结论POP患者宫骶韧带成纤维细胞中WNT16B表达水平低,可能导致细胞的生长增殖活性被抑制以及分泌胶原的能力下降,从而引发POP的疾病进展;WNT16B可能是POP的潜在保护因素之一。Objective:To explore the effects of WNT16B on the proliferation and collagen secretion of uterosacral ligament fibroblasts cultured in vitro and the possible mechanism of pelvic organ prolapse(POP).Methods:Four patients with POP-quantification(POP-Q)stage≥Ⅲ(POP group)and two with other benign gynecologic disorders as control(non-POP group)treated in Peking Union Medical College Hospital,from OCT.2018 to Jun.2021 were recruited in this study.Specimens were taken from the uterosacral ligaments,and fibroblasts were cultured and purified.WNT16B overexpression vector was constructed and transfected into primary cultured fibroblasts.The mRNA expression of WNT16B and Collagen I in fibroblasts was detected by RT-PCR.Real-time cell analysis system(RTCA)was used to detect cell proliferation activity.Results:The mRNA expression of WNT16B in POP group was significantly lower than that in non-POP group(P<0.001).After transfection of WNT16B lentivirus into primary cultured fibroblasts from POP group,the doubling time of fibroblasts was shortened(P<0.05),and the collagen I mRNA was also significantly increased(P<0.05).Conclusions:The low expression of WNT16B in fibroblasts of uterosacral ligament in POP patients may be related to the inhibition of cell growth and proliferation as well as the decreased ability of collagen I secretion,which may be one of the potential protective factors of POP.

关 键 词：盆腔器官脱垂 WNT16B 成纤维细胞 慢病毒转染 

分 类 号：R711.23[医药卫生—妇产科学]