环孢素A对人早孕期滋养细胞MMP-9与MMP-2表达的调控作用

MODULATION OF CYCLOSPORIN A ON THE EXPRESSION OF MMP-9 AND MMP-2 OF THE FIRST-TRIMESTER HUMAN TROPHOBLAST CELLS

本研究的目的是探讨环孢素A对人早孕期滋养细胞侵袭能力及基质金属蛋白酶9与2 (matrix metalloproteinase 9 and 2,简称MMP-9与MMP-2)表达的调节作用,为治疗反复自然流产等妊娠疾患提供新的线索。侵袭试验观察CsA对人早孕期滋养细胞侵袭能力的调节作用;RT-PCR与明胶酶谱分析CsA对滋养细胞MMP-9与MMP-2 mRNA及蛋白水平表达的影响;In-cell West- ern检测CsA作用后滋养细胞ERK1/2磷酸化水平。结果发现,1．0μmol/L CsA明显增强滋养细胞侵袭能力,MEK激酶抑制剂U0126可抑制CsA对滋养细胞的促侵袭作用;1．0μmol/L CsA可诱导MMP- 9与MMP-2基因的转录与蛋白分泌;该诱导效应同样可被U0126所阻滞;1．0μmol/L CsA以时间依赖方式促进ERK1/2的磷酸化。结果表明,CsA可激活ERK1/2,通过MAPK/ERK1/2途径促进滋养细胞MMP-9与MMP-2基因的转录与蛋白分泌,从而增强滋养细胞的侵袭能力,对滋养细胞生物学功能具有良性调节作用。

The aim of this study was to explore whether cyclosporine A （CsA） can modulate expression of MMP-9 and MMP-2 and invasion of the first-trimester human trophoblast cells. The invasive ability of the trophoblast cells modulated by CsA was observed by Matrigel invasion assay. The effect of CsA on the transcription and translation of MMP-9 and MMP-2 was determined by Reverse Transcription-Polymerase Chain Reaction （RT-PCR） and Gelatin Zymography. In-cell Western was used to detect the level of phospho-ERK1/2 of trophoblast cells treated with CsA. We found that CsA（1.0μmol/L） could remarkably promote the invasive ability of the first-trimester human trophoblast cells; CsA（1.0μmol/L） could also induce the expression of MMP-9 and MMP-2 at transcriptional and translational level; U0126,a MEK inhibitor,could obviously inhibit the enhanced invasion and expression of MMP-9 and MMP-2 of these cells induced by CsA;CsA （1.0μmol/L） could activate the ERK1/2 in a time-dependent manner. These results above indicate that CsA can induce the expression of MMP-9 and MMP-2 through activating MAPK/ERK1/2, which contributes to the improved invasion of the first-trimester human trophoblast cells.