白芍总苷通过调控SMAD4/AXL/HIF-1α通路抑制子宫内膜间质细胞迁移、侵袭和上皮间质转化  

Total glucoside of paeony inhibits migration,invasion and epithelial-mesenchymal transition of endometrial stromal cells by regulating SMAD4/AXL/HIF-1αpathway

作　　者：于淼淼 杨露 黄媛媛 崔向华 Yu Miaomiao;Yang Lu;Huang Yuanyuan;Cui Xianghua(Department of Obstetrics and Gynecology,Chinese People’s Liberation Army Eastern Theater Command General Hospital,Nanjing,Jiangsu 210003;Department of Obstetrics and Gynecology,the Fourth Affiliated Hospital of Nanjing Medical University,Nanjing 210031,China)

机构地区：[1]中国人民解放军东部战区总医院妇产科,南京2210003 [2]南京医科大学第四附属医院妇产科,南京210031

出　　处：《中国组织化学与细胞化学杂志》2024年第6期563-570,共8页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：目的探讨白芍总苷(total glucoside of paeony,TGP)介导受体调节型蛋白SMAD家族成员4(SMAD4)对子宫内膜间质细胞迁移、侵袭和上皮间质转化的影响及作用机制。方法用低(0.5 mg/m L)、中(1.0 mg/m L)或高(1.5mg/m L)浓度的TGP处理子宫内膜间质细胞,通过转染pc DNA3.1-SMAD4或sh-SMAD4改变子宫内膜间质细胞SMAD4基因表达;应用qRT-PCR和免疫荧光法检测细胞SMAD4的表达水平,通过蛋白免疫印迹检测SMAD4/酪氨酸蛋白激酶受体(AXL)/低氧诱导因子-1α(HIF-1α)信号通路蛋白与钙黏蛋白(N-cadherin)、波形蛋白(vimentin)等上皮间质转化相关蛋白水平;细胞划痕实验检测细胞迁移能力,Transwell法评估细胞侵袭能力。结果qRT-PCR和免疫荧光检测显示,TGP浓度依赖性降低SMAD4m RNA和蛋白表达;高浓度TGP处理显著降低AXL、HIF-1α、N-cadherin和vimentin的表达,并抑制细胞迁移和侵袭能力,SMAD4过表达显著减弱高浓度TGP处理引起的上述效应,SMAD4的抑制则显著增强高浓度TGP处理引起的上述效应。结论白芍总苷可抑制子宫内膜间质细胞的迁移、侵袭和上皮间质转化,其作用可能与介导SMAD4/AXL/HIF-1α通路有关。Objective To investigate the effects and mechanism of total glucoside of paeony(TGP)in mediating the role of SMAD family member 4(SMAD4)in regulating migration,invasion,and epithelial-mesenchymal transition(EMT)of endometrial stromal cells.Methods Endometrial stromal cells were treated with low(0.5 mg/mL),medium(1.0 mg/mL),or high(1.5 mg/mL)concentrations of TGP.SMAD4 gene expression was altered by transfecting pcDNA3.1-SMAD4 or sh-SMAD4 plasmids into the cells.qRT-PCR and immunofluorescence were used to detect SMAD4 expression levels,and Western blotting was employed to assess the expression of SMAD4,AXL(tyrosine kinase receptor),HIF-1α(hypoxia-inducible factor-1α),and EMT-related proteins such as N-cadherin and vimentin.Cell migration was assessed by scratch assay,and cell invasion was evaluated using Transwell assay.Results qRT-PCR and immunofluorescence showed that TGP concentration-dependently decreased mRNA and protein levels of SMAD4.High-dose TGP treatment significantly reduced the expression of AXL,HIF-1α,N-cadherin,and vimentin,and inhibited cell migration and invasion.SMAD4 overexpression significantly attenuated the effects induced by high-dose TGP treatment,while SMAD4 inhibition significantly enhanced these effects.Conclusion TGP can inhibit migration,invasion,and epithelial-mesenchymal transition of endometrial stromal cells,and its action may be mediated through the SMAD4/AXL/HIF-1αsignaling pathway.

关 键 词：白芍总苷 子宫内膜间质细胞 上皮间质转化 细胞迁移 细胞侵袭 

分 类 号：R711.71[医药卫生—妇产科学]