环孢素A通过活化粘着斑激酶信号通路促进人滋养细胞的体外生长

Cyclosporin A Promotes the Growth of Human Trophoblast Cells In Vitro via Focal Adhesion Kinase Signaling Pathway

目的：探讨粘着斑激酶（focal adhesion kinase，FAK）信号通路在环孢素A（cyclosporin A，CsA）调节人滋养细胞体外生长中的作用。方法：收集因非意愿妊娠6-9周要求行人工流产术的正常妊娠妇女的绒毛组织，体外分离培养获得滋养细胞，经CsA和／或FAK抑制剂Y15处理后。免疫荧光检测滋养细胞FAK的磷酸化水平；BrdU检测滋养细胞的增殖；AnnexinV／PI检测滋养细胞的凋亡。结果：CsA可明显提高滋养细胞FAK的活化水平，FAK抑制剂Y15可阻断CsA诱导的FAK活化，阻抑CsA对细胞增殖的促进作用（P〈0．05）以及对滋养细胞凋亡的抑制作用（P〈0．05）。结论：CsA可通过活化FAK信号通路促进人早孕期滋养细胞的体外生长，从而有利于正常妊娠的维持。

Objective： To investigate the role of focal adhesion kinase （FAK） signaling pathway on cyclosporin A （CsA）-induced growth of human trophoblast cells in vitro. Methods： Human villous tissues were collected from elective termination of first trimester pregnancies （6-9 weeks of gestation） for non-medical reasons. Primary trophoblast cells were prepared by the trypsin-DNase I digestion. After treatments of trophoblast cells with CsA and/or Y15 （a specific inhibitor of FAK）, the phosphorylation of FAK was detected by immunofluorescence assay, the proliferation was examined by BrdU incorporation, and the apoptosis was evaluated by Annexin V/PI staining. Results： CsA significantly enhanced the activation of FAK in human primary trophoblasts, and Y15 inhibited this activity of CsA. In addition, Y15 abrogated CsA-increased proliferation and CsA-decreased apoptosis of trophoblast cells. Conclusion： CsA promotes the growth Of human trophoblasts in vitro via FAK signaling pathway which is involved in the favorable modulation of CsA on pregnancy maintenance.