GPR30蛋白对滋养细胞功能的影响与子痫前期发病的关系

Effect of GPR30 protein on function of trophoblast cells and its relationship with the development of preeclampsia

目的:探讨G-蛋白偶联受体30(G-protein coupled receptor 30,GPR30)在不同妊娠状态时的滋养细胞上的表达情况,以及其表达变化与子痫前期(preeclampsia,PE)发病的关系。方法:应用免疫荧光及免疫组化的方法检测不同孕周早孕期绒毛中GPR30的表达,应用免疫组化及Western blot检测晚孕期正常和PE患者胎盘组织中GPR30的表达水平。用缺氧复氧(hypoxiareoxygenation,H/R)处理人类绒毛膜滋养层细胞株HTR8/SVneo以建立PE模型,检测其GPR30的表达状况。用17β-雌二醇(17β-estradiol,E2)、GPR30特异性激动剂G1和阻滞剂G15处理HTR8/SVneo细胞及绒毛外植体,观察处理因素对滋养层细胞侵袭力的影响。结果:GPR30在早期绒毛组织细胞滋养细胞上表达水平高;晚孕期正常胎盘滋养细胞上GPR30的表达明显高于PE胎盘。在PE细胞模型上GPR30表达明显减少。E2及G1可增加滋养细胞的侵袭能力,G15则可下调其侵袭力。结论:GPR30表达降低可能下调滋养细胞侵袭力,从而与PE发病有关。

Objective：To characterize the expression of G-protein coupled receptor 30（GPR30）in trophoblast cells at different conditions,and to determine the effect of disordered expression of GPR30 on the development of preeclampsia（PE）. Methods：The expressions of GPR30 protein were detected not only in villi at first trimester,placentas from normal term and pregnancies complicated with PE,but also in HTR8/SVneo cells which treated with normoxia and hypoxia-reoxygenation（H/R）by immunofluorescence,immunohistochemistry and/or Western blot. The pharmacological modulations of GPR30 were built by the using of 17 β-estradiol（E2）,GPR30 specific agonist G1 and GPR30 specific inhibitor G15. H/R was applied to simulate the pathological condition of PE;their effect on human trophoblast cell invasion was determined using vitro invasion assays. Results：GPR30 protein was highly expressed in the cytotrophoblast of first trimester villi and the term placentas from the gestational matched controls（GMCs）,but significantly compromised in PE cells model and placentas which complicated by PE. In addition,E2 and G1 treatment significantly increased the invasive capacity of trophoblast cells,however,the antagonist G15 play an opposite role. Meanwhile,H/R significantly attenuated invasiveness of trophoblast cells. Conclusion：the pathogenetic mechanism of PE is associated with the downregultaion of GPR30 protein level in trophoblast which may impair the invasive capacity of trophoblast cells.