CyPA interacts with SERPINH1 to promote extracellular matrix production and inhibit epithelial-mesenchymal transition of trophoblast via enhancing TGF-β/Smad3 pathway in preeclampsia
CyPA interacts with SERPINH1 to promote extracellular matrix production and inhibit epithelial-mesenchymal transition of trophoblast via enhancing TGF-β/Smad3 pathway in preeclampsia
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DOI：
10.1016/j.mce.2022.111614
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发表时间：
2022-03-21
影响因子：
4.1
通讯作者：
Zhong, Mei
中科院分区：
文献类型：
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作者：
Hu, Haoyue;Ma, Jing;Zhong, Mei
我们先前报道了亲环素A（CyPA）的产生在先兆子痫（PE）中上调。此外，已知CyPA在妊娠小鼠中诱导PE样特征并损害滋养层侵袭性。在本研究中，我们进一步阐明了CyPA在PE中的作用。小鼠胎盘的RNA-seq分析、RT-qPCR、免疫组织化学（IHC）染色和蛋白质印迹显示，CyPA增加细胞外基质（ECM）蛋白（如胶原蛋白I和纤连蛋白）的水平，并激活TGF-13/Smad 3信号通路。此外，CyPA抑制参与上皮-间充质转化（EMT）的基因的表达（例如，E-钙粘蛋白、N-钙粘蛋白和波形蛋白）。然后，我们构建了稳定的过表达和敲低CyPA的细胞模型（使用HTR 8/SVneo细胞），以阐明分子机制。我们发现CyPA通过TGF-13/Smad 3途径调节ECM相关蛋白的水平和EMT过程。我们还确定SERPINH 1作为一个假定的CyPA结合蛋白，使用液相色谱-电喷雾质谱（LC-MS）/MS。SERPINH 1被发现在PE胎盘上调。沉默SERPINH 1表达逆转了ECM蛋白的上调和CyPA过表达诱导的EMT过程的抑制。这些发现揭示了CyPA在PE中滋养细胞侵袭力受损中的功能，并表明CyPA和SERPINH 1可能是治疗PE的有希望的靶点。
We previously reported that cyclophilin A (CyPA) production is upregulated in preeclampsia (PE). Moreover, CyPA is known to induce PE-like features in pregnant mice and impair trophoblast invasiveness. In this study, we further illustrated the role of CyPA in PE. RNA-seq analysis, RT-qPCR, immunohistochemical (IHC) staining, and western blotting of mouse placentae revealed that CyPA increased the levels of extracellular matrix (ECM) proteins, such as collagen I and fibronectin, and activated the TGF-13/Smad3 signaling pathway. Additionally, CyPA inhibited the expression of genes involved in epithelial-mesenchymal transition (EMT) (e.g., E-cadherin, N-cadherin, and vimentin) in mouse placentae. We then constructed stable overexpressing and knock-down CyPA cell models (using HTR8/SVneo cells) to clarify the molecular mechanism. We found that CyPA regulated the levels of ECM-related proteins and the EMT process through the TGF-13/Smad3 pathway. We also identified SERPINH1 as a putative CyPA-binding protein, using liquid chromatography-electrospray mass spectrometry (LC-MS)/MS. SERPINH1 was found to be upregulated in the placentae of PE. Silencing SERPINH1 expression reversed the upregulation of ECM proteins and inhibition of the EMT process induced by the overexpression of CyPA. These findings revealed the functions of CyPA in the impaired invasiveness of trophoblasts in PE and indicated that CyPA and SERPINH1 may represent promising targets for the treatment of PE.