缺氧对体外培养的早孕期细胞滋养细胞中缺氧适应相关部分基因表达及细胞浸润能力的调控

Regulation of hypoxia on the expression of HIF-1α,HPH/PHD1,HPH/PHD2 and FIH-1 in cultured cytotrophoblast of first trimester in vitro and in cell's infiltration ability

目的:探讨缺氧对细胞滋养细胞表达缺氧诱导因子-1α(HIF-1α)及氧感受器缺氧诱导因子脯氨酸羟化酶1、2(HPH/PHD1、2)和抑制缺氧诱导因子-1(FIH-1)的调控,以及缺氧对体外培养的细胞滋养细胞浸润能力的调节。方法:用RT-PCR和Western blot法检测在常氧、缺氧再复氧及持续缺氧等条件下,细胞滋养细胞中HIF-1α、HPH/PHD1、2和FIH-1 mRNA和蛋白表达的变化;并用体外侵袭实验检测缺氧再复氧、持续低氧对细胞滋养细胞浸润能力的影响。结果:(1)持续缺氧条件下培养的细胞滋养细胞HIF-1α和HPH/PHD2 mRNA和蛋白表达显著高于常氧组(P<0.01);HPH/PHD1和FIH-1 mRNA和蛋白表达显著低于常氧组(P<0.01)。缺氧再复氧后培养的细胞滋养细胞HIF-1α和HPH/PHD2 mRNA和蛋白表达显著高于常氧组(P<0.05),但显著低于持续缺氧组(P<0.05);HPH/PHD1和FIH-1 mRNA和蛋白表达均显著低于常氧组(P<0.05),但显著高于持续缺氧组(P<0.05)。(2)持续缺氧显著抑制细胞滋养细胞的浸润能力,与常氧组比较明显降低(P<0.01);缺氧后再复氧细胞滋养细胞的浸润能力居中,与常氧组比较明显降低(P<0.05);与持续缺氧组比较明显升高(P<0.05)。结论:滋养细胞可感受氧分压变化,HIF-1α、HPH/PHD1、HPH/PHD2和FIH-1相互作用与滋养细胞浸润能力调节有关。

Objective ：To investigate the influence of hypoxia on the expression of HIF-1α, HPH/PHDI, HPH/PHD2 and FIH-1 in the cultured cytotrophoblast of first trimester in vitro, and the regulation of hypoxia on the infiltration ability of eytotrophoblast cultured in vitro. Methods：RT-PCR and Western blot were used to detect the mRNA and protein expressions of HIF-1α, HPH/PHD1, HPH/PHD2 and FIH-1 in the cytotrophoblast which cultured in vitro under normal oxygen, consistent hypoxia, and the environment which firstly in hypoxia then in normal oxygen. Infiltration test was used to detect the influence of consistent hypoxia and hypoxiathen-normal oxygen to infiltration ability of cytotrophoblasts cultured in vitro. Results： When cultured in the consistent hypoxia, the mRNA and protein expressions of HIF-1α and HPH/ PHD2 were obviously higher then normal oxygen （P 〈 0.01 ）. The mRNA and protein expressions of HPH/PHD1 and FIH-1 were obviously lower （P 〈 0.01 ）. When cuhured in hypoxia firstly then in normal oxygen, the mRNA and protein expressions of HIF-1α and HPH/PHD2 were obviously higher than that cultured in normal oxygen, but lower than that cultured in consistent hypoxia （P 〈0.05 ,P 〈 0.05 ）;the mRNA and protein expressions of HPH/PHD1 and FIH-1 were obviously lower than that cultured in normal oxygen, but higher than that cultured in consistent hypoxia （ P 〈 0.05, P 〈 0.05 ）. The infiltration ability of cytotrophoblast cultured in vitro was inhibited by hypoxia-then-normal hypoxia;and more obviously inhibited by consistent hypoxia. Conclusion：The hypoxia regulates the infiltration ability of cytotrophoblast maybe through the regulation on HIF-1α,HPH/PHD1 ,HPH/PHD2 and FIH-1.