WNK1基因对宫骶韧带成纤维细胞生长的影响  

Effect of WNK 1 gene on growth of fibroblasts in the sacral ligament

作　　者：乔艳华 赵冰[1] 李林玉 王莹莹 王肖帆 张志磊 孙青 QIAO Yan-hua;ZHAO Bing;LI Lin-yu;WANG Ying-ying;WANG Xiao-fan;ZHANG Zhi-lei;SUN Qing(Department of Obstetrics and Gynecology,the Third Affiliated Hospital of Zhengzhou University,Zhengzhou Henan 450001;Ministry of Science and Education,Sanquan College of Xinxiang Medical College,Xinxiang Henan 453003,P.R.China)

机构地区：[1]郑州大学第三附属医院妇产科,河南郑州450001 [2]新乡医学院三全学院科教部,河南新乡453003

出　　处：《中国计划生育和妇产科》2020年第4期89-91,96,共4页Chinese Journal of Family Planning & Gynecotokology

基　　金：河南省高校科技创新人才支持计划(项目编号:18HASTIT045)。

摘　　要：目的研究WNK 1基因对宫骶韧带成纤维细胞的影响。方法采用组织块法培养大鼠宫骶韧带成纤维细胞。根据慢病毒转染的方法分为WNK 1+组、WNK 1-组和空白对照组。RT-qPCR检测细胞的转染效率;CCK-8法检测细胞增殖的情况;用RT-qPCR和Western blot检测成纤维细胞WNK 1基因、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白表达。结果WNK 1+组的成纤维细胞的WNK 1基因表达明显高于WNK 1-组及空白对照组(P<0.05);镜检及CCK-8检测成纤维细胞增殖数目和体积均显著增加(P<0.05);Western-blot及RTq PCR结果显示WNK 1+组WNK 1激酶、Ⅰ型和Ⅲ型胶原蛋白的mRNA相对表达量及蛋白水平较WNK 1-组及空白对照组均显著增加(P<0.05)。结论WNK 1基因能够促进宫骶韧带中成纤维细胞的增殖,使胶原蛋白表达增加,继而能增加宫骶韧带的弹力及张力,为患有盆底功能障碍疾病患者的诊断和治疗提供新思路。Objective To study the effect of WNK 1 gene on fibroblasts of uterine sacral ligament.Methods Tissue block method was used to culture rat uterine sacral ligament fibroblasts.They were divided into WNK 1+group,WNK 1-group and blank control group according to the method of lentivirus transfection.RT-qPCR was used to detect cell transfection efficiency;CCK-8 method was used to detect cell proliferation;RT-qPCR and Western blot were used to detect the expression of WNK 1 gene,typeⅠcollagen,and typeⅢcollagen in fibroblasts.Results The WNK 1 gene expression of fibroblasts in WNK 1+group was significantly higher than that in WNK 1-group and blank control group(P<0.05);the number and volume of fibroblast proliferation were significantly increased under microscopic examination and CCK-8 detection(P<0.05).Western-blot and RT-qPCR results showed that the relative expression levels and protein levels of WNK 1 kinase,typeⅠand typeⅢcollagen in WNK 1+group were significantly higher than those in WNK 1-group and blank control group(P<0.05).Conclusion WNK 1 gene can promote the proliferation of fibroblasts in the uterine sacral ligament,increase collagen expression,and then increase the elasticity and tension of the uterine sacral ligament,providing new ideas for diagnosis and treatment of patients with pelvic floor dysfunction.

关 键 词：WNK1基因 宫骶韧带 成纤维细胞 I型胶原蛋白 Ⅲ型胶原蛋白 

分 类 号：R711.2[医药卫生—妇产科学]