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目的:探讨缺氧对滋养细胞可溶性血管内皮生长因子受体1(soluble fims-like tyrosine kinase receptor-1,sFlt-1)及血管内皮生长因子(vascular epithelial growth factor,VEGF)表达的影响。方法:CoCl2诱导人早孕绒毛滋养细胞系TEV-1化学缺氧,分别于0h、24h、48h、72h、96h、120h用Real-time RT-PCR法检测滋养细胞中sFlt-1及VEGF mRNA表达,ELISA法检测细胞培养上清液中sFlt-1及VEGF蛋白表达。结果:滋养细胞缺氧72h后,胞质内可见明显空泡。滋养细胞sFlt-1mRNA及蛋白表达随缺氧时间的延长逐渐升高,且与缺氧72h、96h、120h间差异有显著性(P<0.05)。而滋养细胞VEGF mRNA及蛋白表达随缺氧时间的延长呈现出先上升后下降的趋势,并于缺氧48h升高达到峰值(P<0.05)。结论:不同的缺氧时间对滋养细胞VEGF及sFlt-1表达具有重要的调控作用。慢性缺氧可能导致滋养细胞sFlt-1及VEGF表达平衡失调,从而参与子痫前期的发生、发展。
Objective: To investigate the effect of hypoxia on the expression of soluble fims-like tyrosine kinase receptor-1 (sFlt-1) and vascular epithelial growth factor (VEGF) in trophoblast cells. Methods: CoCl2 induced the expression of sFlt-1 and VEGF mRNA in gestational trophoblast cell line TEV-1. Real-time RT-PCR was used to detect the expression of sFlt-1 and VEGF at 0h, 24h, 48h, 72h, 96h and 120h respectively. ELISA was used to detect the expression of sFlt-1 and VEGF in the cell culture supernatant. Results: After hypoxia for trophoblast 72h, obvious vacuoles were found in cytoplasm. The expression of sFlt-1 mRNA and protein in trophoblast cells increased gradually with the prolongation of hypoxia, and there was a significant difference between 72h, 96h and 120h hypoxia (P <0.05). However, the expression of VEGF mRNA and protein in trophoblast cells increased first and then decreased with the prolongation of hypoxia, and peaked at 48h (P <0.05). CONCLUSION: Different hypoxic periods play an important regulatory role on the expression of VEGF and sFlt-1 in trophoblasts. Chronic hypoxia may lead to dysregulation of sFlt-1 and VEGF expression in trophoblasts and thus participate in the development of preeclampsia.