NF-κB信号通路在人羊膜间充质干细胞治疗宫腔粘连中的作用

Role of NF-κB signaling pathway in treatment of intrauterine adhesions by human amniotic mesenchymal stem cells

目的探讨IκB-α/NF-κB信号通路在人羊膜间充质干细胞(human amniotic mesenchymal stem cells,hAMSCs)治疗宫腔粘连(intrauterine adhesion, IUA)中的作用机制。方法体内实验:18只SD大鼠分为Sham组、IUA模型组及hAMSCs组;采用HE及Masson染色分别计数子宫内膜腺体个数及纤维化面积百分比,免疫组化法检测子宫内膜组织中IκB-α(NF-κB抑制蛋白)、核转录因子P65、E-型钙粘蛋白(E-cadherin)、波形蛋白(Vimentin)表达水平。体外实验:分别用外源性生长因子[10 ng/mL(TGF-β1+EGF+PDGF-BB),诱导组]及10 nmol/L BAY11-7082联合生长因子[10 ng/mL(TGF-β1+EGF+PDGF-BB),抑制剂组]诱导hAMSCs 5 d,以不加生长因子诱导的hAMSCs作为对照组;RT-PCR检测各组中IκB-α、角蛋白(CK-7、CK-19)的表达水平。结果体内实验:IUA模型组中IκB-α、P65、Vimentin较Sham组表达增加,E-cadherin表达下降,差异均有统计学意义(P<0.05);hAMSCs组中IκB-α、P65、Vimentin较IUA模型组表达下降,E-cadherin表达增加,差异均有统计学意义(P<0.05)。体外实验:诱导组中IκB-α较对照组表达下降,CK-7表达增加;抑制剂组中CK-7、CK-19较诱导组明显增加,差异均有统计学意义(P<0.05)。结论 hAMSCs移植具有治疗宫腔粘连的作用,其作用机制可能是通过抑制IκB-α/NF-κB通路激活从而促进hAMSCs向子宫内膜上皮细胞转化。

Objective To investigate the role of IκB-α/NF-κB signaling pathway in the treatment of intrauterine adhesions(IUA) by human amniotic mesenchymal stem cells(hAMSCs). Methods In vivo experiment: 18 SD female rats were randomly divided into Sham group, IUA model group and hAMSCs group. The number of endometrial glands and the percentage of fibrotic area were calculated by HE staining and Masson staining, respectively. Immunohistochemistry was used to detect the expression levels of IκB-α(NF-κB inhibitor), nuclear transfer factor P65, E-cadherin and vimentin in endometrial tissue. In vitro experiments: hAMSCs were induced by exogenous growth factors(10 ng/mL TGF-β1+EGF+PDGF-BB) and 10 nmol/L BAY11-7082 combined with above growth factors for 5 d, respectively, and the cells without treatment served as blank control. Fluorescence quantitative PCR was used to detect the expression levels of IκB-α, and cytokeratins CK-7 and CK-19. Results In vivo experiment: The expression levels of IκB-α, P65 and vimentin was significant higher, while that of E-cadherin was obviously lower in the model group than the Sham group(all P<0.05);the expression levels of IκB-α, P65 and vimentin in the hAMSCs group were notably lower, while that of E-cadherin was higher than the model group(all P<0.05). In vitro experiment: Compared with the control group, the expression of IκB-α was decreased but that of CK-7 was increased in the induction group;The expression levels of CK-7 and CK-19 were increased significantly in the inhibitor group than the induction group(both P<0.05). Conclusion hAMSCs transplantation have therapeutic effect on uterine adhesions, which may be due to its promoting the transformation of hAMSCs into endometrial epithelial cells by inhibition of IκB-α/NF-κB pathway activation.