月经发生中低氧诱导因子-1A对VEGF的调节

The regulation of VEGF expression by hypoxia-inducible factor-1A during menstruation

目的在小鼠月经样模型和人蜕膜化子宫内膜基质细胞模拟月经发生基础上,探索低氧诱导因子-1A(HIF1A)对血管内皮生长因子(VEGF)的表达调控作用,以及月经发生中HIF1A对VEGF mRNA表达的调控机制。方法建立小鼠月经样模型,用ChIP方法检测HIF1A是否直接调控Vegf mRNA的表达;利用HIF1A抑制剂甲氧雌二醇(2ME)抑制HIF1A的功能,检测小鼠月经样模型中HIF1A、VEGF蛋白的表达量;人子宫内膜基质细胞体外诱导蜕膜化并模拟月经发生,检测HIF1A与VEGF mRNA的表达;用siRNA敲降的方法抑制HIF1A,检测VEGF mRNA的表达。结果小鼠月经样模型在孕酮撤退0h,HIF1A结合的Vegf promoter的相对百分比较低,随后在8h和12h逐渐上升,在12h达到峰值(P<0.05);2ME抑制HIF1A入核的同时,抑制了VEGF的表达量(P<0.05);人蜕膜化子宫内膜基质细胞中,孕酮撤退显著上调HIF1A和VEGF mRNA的表达量(P<0.01);敲降HIF1A后,VEGF mRNA的上调受到明显的抑制(P<0.01)。结论在月经发生中,孕酮撤退能够激活HIF1A转录因子的功能,HIF1A被激活后,结合于VEGF promoter区并启动VEGF mRNA的表达。

Objective：To investigate the regulation of HIF1 A（hypoxia-inducible factor）on the expression of vascular endothelial growth factor（VEGF）protein and mRNA in mouse menstrual models and in vitro cultured human decidualized endometrial stromal cells to explore the role of HIF1 Ain menstruation.Methods：The physiological menstruation model of progesterone withdrawal in mouse was established and ChIP assay was used to detect the direct regulation of HIF1 A on Vegf.HIF1 A inhibitor methoxyestradiol（2 ME）was used to intervene in the menstrual-like model and detect the expression of HIF1 Aand VEGF protein.Human decidualized endometrial stromal cells were prepared in vitro to simulate menstruation.The expression of VEGF mRNA was detected after knockdown of HIF1 A by siRNA.Results：In the mouse menstrual-like model at 0 h after progesterone withdrawal,the relativepercentage of HIF1 A-conjugated Vegf promoter was lower,then it gradually increased at 8 hand 12 h,peaked at 12 h（P〈0.05）.At 16 h,the relative percentage of HIF1 A-conjugated Vegf promoter rapidly decreased and increased slightly at 24 h,but only about 1/2 of the peak level at 12 h.2 ME inhibited the entry of HIF1 Ainto the nucleus and significantly decreased VEGF expression（P〈0.05）.In human decidualized endometrial stromal cells,progesterone withdrawal significantly up-regulated the expression of HIF1 Aand VEGF mRNA.Knockdown of HIF1 Asignificantly inhibited the expression of VEGF mRNA（P〈0.01）.Conclusions：In the onset of menstruation after the progesterone withdrawal,HIF1 Ais translocated to the nucleus and transcription factor function activated,and the expression of the downstream regulatory gene VEGF is directly initiated.