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目的:探讨G-蛋白偶联受体30(GPR30)在妊娠期胎盘中的表达,以及其对人胎盘滋养细胞侵袭力的影响。方法:免疫组化法检测早孕期绒毛、正常产妇胎盘和重度子痫前期患者胎盘组织中GPR30表达。用17-β-雌二醇(17β-E2)、GPR30激动剂G1和阻滞剂G15预处理体外培养绒毛组织和人绒毛外滋养细胞株HTR8/SVneo。显微镜下观察体外绒毛组织滋养细胞生长侵袭范围,Transwell侵袭实验检测HTR8/SVneo细胞侵袭能力。免疫荧光法检测绒毛组织和HTR8/SVneo细胞中GPR30蛋白表达。Western blot法检测HTR8/SVneo细胞中GPR30和MMP-9蛋白表达。结果:GPR30在早期绒毛组织和正常末期胎盘滋养细胞上都有表达,且早孕期的表达水平高于正常末期胎盘,但在重度子痫前期胎盘上GPR30蛋白表达明显减少。E2及GPR30激动剂G1可增加滋养细胞的侵袭能力,阻滞剂G15则可下调其侵袭性;E2、G1可诱导滋养细胞GPR30蛋白表达上调,而G15则下调其表达。GPR30蛋白水平与侵袭相关蛋白MMP-9表达水平有相关性。结论:GPR30可能参与人类滋养细胞侵袭力的调节,对滋养细胞的侵袭力有正性促进作用。
Objective: To investigate the expression of G-protein coupled receptor 30 (GPR30) in gestational placenta and its effect on human placental trophoblast invasion. Methods: The expression of GPR30 in placenta of early pregnancy villus, normal maternal placenta and severe preeclampsia was detected by immunohistochemistry. In vitro cultured chorionic villus and human chorionic villus cell line HTR8 / SVneo were pre-treated with 17-β-estradiol (17β-E2), GPR30 agonist G1 and blocker G15. The growth of trophoblast cells in vitro was observed under a microscope, and the invasion ability of HTR8 / SVneo cells was detected by Transwell invasion assay. Immunofluorescence method was used to detect GPR30 protein expression in villi and HTR8 / SVneo cells. Western blot was used to detect the expression of GPR30 and MMP-9 in HTR8 / SVneo cells. Results: GPR30 was expressed in early villi and normal end-stage placental trophoblasts, and the expression level of GPR30 in early pregnancy was higher than that in normal end-stage placenta. However, GPR30 protein expression in severe preeclampsia placenta was significantly decreased. E2 and GPR30 agonist G1 can increase the invasion ability of trophoblast cells, and blocker G15 can down-regulate its invasiveness. E2 and G1 can up-regulate the expression of GPR30 protein in trophoblast cells, while G15 down-regulates its expression. The GPR30 protein level is correlated with the expression of MMP-9. Conclusion: GPR30 may be involved in the regulation of human trophoblast invasiveness, which has a positive effect on trophoblast invasiveness.