基于SHH-Gli通路调控EMT探讨补肾活血方防治宫腔重度粘连TCRA分离术后复发的机制  

To investigate the mechanism of Bushen Huoxue Recipe in preventing and treating recurrence of severe intrauterine adhesions after TCRA separation based on SHH-Gli pathway regulating EMT

作　　者：罗梅[1,2] 闻洪丽 邹凌凤 汤锶锶[1,2] 夏敏 LUO Mei;WEN Hong-li;ZOU Ling-feng;TANG Si-si;XIA Min(Chongqing Traditional Chinese Medicine Hospital,Chongqing,400021,China;The First Clinical College,Chongqing College of Traditional Chinese Medicine,Chongqing 400021,China)

机构地区：[1]重庆市中医院,重庆400021 [2]重庆中医药学院第一临床学院,重庆400021

出　　处：《时珍国医国药》2024年第11期2523-2529,共7页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金青年基金项目(82004414);成都中医药大学2021年度“杏林学者”医院专项(YYZX2021053);重庆市自然科学基金(CSTB2022NSCQ-MSX1124);重庆市中青年医学高端人才项目([2022]15号)。

摘　　要：目的通过体外实验探究补肾活血方抑制宫腔粘连患者TCRA术后复发的机制。方法通过TGF-β1诱导建立子宫内膜间质细胞纤维化模型,将细胞分为空白对照组、TGF-β1诱导组、TGF-β1+SHH抑制组、TGF-β1+阴性血清组、TGF-β1+含药血清组、TGF-β1+含药血清+pcDNA-NC组和TGF-β1+含药血清+pcDNA-SHH组共7组。采用CCK8和细胞划痕实验分别检测细胞的增殖及迁移能力;采用ELISA检测细胞上清中Ⅰ型胶原α1链(collagen typeⅠα1 chain,COL1A1)分泌水平;采用免疫组化检测细胞中E-钙黏蛋白(E-cadherin)及波形蛋白(Vimentin)的表达;采用RT-qPCR检测细胞中SHH-Gli信号相关因子的表达。结果与TGF-β1诱导组相比,TGF-β1+SHH抑制组和TGF-β1+含药血清组的细胞增殖及迁移力、COL1A1分泌水平、Vimentin表达及SHH、Gli1、Gli2的mRNA含量降低(P<0.05),E-cadherin的表达增加(P<0.05)。与TGF-β1+含药血清+pcDNA-NC组比较,TGF-β1+含药血清+pcDNA-SHH组细胞增殖及迁移力、COL1A1分泌水平、Vimentin表达及SHH、Gli1、Gli2的mRNA含量增加(P<0.05),E-cadherin的表达降低(P<0.05)。结论补肾活血方含药血清能够通过抑制SHH-Gli通路相关基因的表达,从而减少基质细胞向成纤维细胞分化,减轻ESCs纤维化。Objective To investigate whether serum containing Bushen Huoxue Recipe inhibits the process of recurrent endometrial fibrosis after TCRA in patients with intrauterine adhesions in vitro,and to clarify the role of SHH-Gli signaling pathway-related molecule expression in this process.Methods Fibrotic cell models were established by inducing fibrosis of endometrial stromal cells with TGF-β1,and the cells were divided into seven groups:control group,TGF-β1 induced group,TGF-β1+SHH inhibited group,TGF-β1+negative serum group,TGF-β1+medicated serum group,TGF-β1+medicated serum+pcDNA-NC group,and TGF-β1+medicated serum+pcDNA-SHH group.CCK8 and scratch assay were used to detect the proliferation and migration of cells,respectively;ELISA was used to detect the secretion level of collagen type Iα1 chain(COL1A1)in the cell supernatant;immunohistochemistry was used to detect the expression of E-cadherin and vimentin in the cells;RT-qPCR was used to detect the expression of SHH-Gli signaling-related factors in the cells.Results Compared with the TGF-β1 induced group,cell proliferation and migration,COL1A1 secretion level,vimentin expression and mRNA content of SHH,Gli1,and Gli2 were decreased(P<0.05),and the expression of E-cadherin was increased(P<0.05)in the TGF-β1+SHH inhibited and TGF-β1+medicated serum groups.Compared with the TGF-β1+medicated serum+pcDNA-NC group,the TGF-β1+medicated serum+pcDNA-SHH group had increased cell proliferation and migration,COL1A1 secretion level,vimentin expression and mRNA content of SHH,Gli1,and Gli2(P<0.05),and decreased expression of E-cadherin(P<0.05).Conclusion Bushen Huoxue Recipe medicated serum can reduce the differentiation of stromal cells into fibroblasts and reduce ESCs fibrosis by inhibiting the expression of SHH-Gli pathway-related genes.

关 键 词：子宫内膜间质细胞 补肾活血方 TGF-β1 SHH-Gli信号 宫腔粘连 

分 类 号：R285.5[医药卫生—中药学]