转化生长因子-β1/Smad 3在机械力下调盆底成纤维细胞胶原、弹性蛋白表达中的作用  

The function of TGF-β1/Smad 3 pathway on the expression of collagen and elastin reduced by mechanical strain in pelvic fibroblast cells

作　　者：李秉枢[1] 洪莉[1] 程丽薇[1] 杨将[1] 李素廷 何松明[1] 汤建明[1] LI Bing-shu;HONG Li;CHENG Li-wei;YANG Jiang;LI Su-ting;HE Song-ming;TANG Jian-ming(Department of Gynecology and Obstetrics,Reople＇s Hospital of Wuhan University,Wuhan Hubei 430060,P.R.China)

机构地区：[1]武汉大学人民医院妇产科,湖北武汉430060

出　　处：《中国计划生育和妇产科》2018年第10期54-58,共5页Chinese Journal of Family Planning & Gynecotokology

基　　金：湖北省自然科学基金(项目编号:2017CFB750);中央高校基本科研业务费专项资金(项目编号:2042016kf0120);国家自然科学基金面上项目(项目编号:81471442)

摘　　要：目的探讨转化生长因子-β1(transforming growth factor-β1,TGF-β1)/Smad 3信号通路在机械力下调盆底成纤维细胞胶原、弹性蛋白表达中的作用。方法收集2013年1月至2015年1月于武汉大学人民医院因非盆腔器官脱垂行阴式全子宫切除患者的骶主韧带组织,原代培养成纤维细胞,根据干预因素不同进行如下分组:(1)对照组:未加力及不添加TGF-β1孵育的细胞爬片细胞;(2)机械力组(cyclic mechanical stretch,CMS组):力学参数4 mm 0. 5 Hz的机械力加载人盆底成纤维细胞4 h;(3) TGF-β1组:10 ng/m L TGF-β1孵育人盆底成纤维细胞24 h;(4) TGF-β1+CMS组:10 ng/m L TGF-β1孵育人盆底成纤维细胞24 h后,力学参数4 mm 0. 5 Hz的机械力加载人盆底成纤维细胞4 h。采用Western blot检测各组中TGF-β1、Smad 2/3(drosophila mothers against decapentaplegic 2/3,Smad 2/3)、磷酸化(phosphorylation,p-) Smad 2、磷酸化(phosphorylation,p-) Smad 3蛋白表达情况。结果与对照组相比,CMS组TGF-β1、p-Smad 3、I型胶原蛋白A1(Collagen 1A1,COL 1A1)、Ⅲ型胶原蛋白A1(Collagen 3A1,COL 3A1)、弹性蛋白(Elastin)表达明显下调,差异均有统计学意义(P <0. 05),Smad 2/3、p-Smad 2蛋白表达差异无统计学意义(P> 0. 05)。TGF-β1+CMS组与TGF-β1组相比,p-Smad 3、COL 1A1、COL 3A1、Elastin蛋白表达明显下调,差异均有统计学意义(P<0. 05),Smad 3蛋白表达差异无统计学意义(P> 0. 05)。与对照组相比,TGF-β1组COL 1A1、COL 3A1、Elastin、p-Smad 3蛋白表达明显上调,差异均有统计学意义(P <0. 05),Smad 3蛋白表达差异无统计学意义(P> 0. 05)。TGF-β1+CMS组与CMS组相比,p-Smad 3、COL 1A1、COL 3A1、Elastin蛋白表达明显上调,差异均有统计学意义(P <0. 05),Smad 3蛋白表达差异无统计学意义(P> 0. 05)。结论机械力下调盆底成纤维细胞内TGF-β1蛋白表达,抑制Smad 3信号通路磷酸化,从而下调COL 1A1、COL 3A1及Elastin蛋白表达;外源性TGF-β1磷酸化激活Smad 3后可修复机械�Objective To investigate the role of transforming growth factor - β1 （ TGF - β1 ）/Smad 3 signaling pathway in down -regulating the expression of collagen and elastin in pelvic floor fibroblasts. Methods The sacral ligament tissues of the vaginal hysterectomy were collected from the non - pelvic organ prolapse patients from People＇s Hospital of Wuhan University from January 2013 to January 2015. The primary cultured fibroblasts were divided into groups according to the intervention factors： ① Control group： 4 mm 4 h 0. 5 Hz（ - ）, TGF - 131 （ - ）. ② Cyclic mechanical stretch （CMS） group：4 mm 4 h 0. 5 Hz（ ＋ ）, TGF - β1 （ - ）. ③ TGF-β1 group： 4mm4 h0. 5 Hz（ -）,TGF-131（＋）. ④TGF-β1＋CMSgroup： 4 mm4 h0.5 Hz（＋）, LY294002（＋）. Western blotting was used to evaluate the expression of TGF - 131, Smad 2/3, p - Smad 2, p - Smad 3, COL1A1, COL3A1 and elastin. Results The results of western blotting showed a significantly lower expression of TGF - β1, p - Smad 3, COL1 A1, COL3A1 and Elastin in CMS group（P 〈0. 05） ,but the expressions of Smad 2/3 and p - Smad 2 had no statistical difference compared to the control group （P 〉 0. 05 ）. The expression of COL1A1, COL3A1, Elastin and p - Smad 3 in TGF - β1 group were higher than the control group （ P 〈 0.05 ）, yet the expression of Smad 3 was not statistical different from the control group （ P 〉 0. 05 ）. The expression of COL1 A1, COL3A1 and Elastin in TGF - β1 ＋ CMS group were higher than the CMS group （ P 〈 0. 05 ）, but the expression of Smad 3 was not statistical different from the control group （ P 〉 0. 05 ）. The expression of p - Smad 3, COL1 A1, COL3 A1, Elastin in TGF - β1 ＋ CMS group were higher than the CMS group （ P 〈 0. 05 ）. However, there was no statistical difference of the expression of Smad 3 （ P 〉 0. 05 ）. Conclusion Mechanical strain could inhibit the phosphorylation of Smad 3, and down - regulate the expression of COL1A1, COL3Aland elasti

关 键 词：盆腔器官脱垂 TGF-Β1/SMAD 3 机械力 胶原蛋白 弹性蛋白 

分 类 号：R711.2[医药卫生—妇产科学]