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目的在小鼠月经模型基础上,研究低氧诱导因子(HIF1A)与血管内皮生长因子(VEGF)蛋白及mRNA的表达与定位,探索HIF1A与VEGF在月经发生中可能的调控机制。方法建立小鼠月经样模型,假孕小鼠通过花生油子宫腔注射的方法诱导小鼠子宫内膜发生蜕膜化,切除卵巢致孕酮(P4)撤退以模拟月经的发生。在P4撤退后0、8、12、16、24h分别收集小鼠子宫;利用免疫组化与Western blot方法检测HIF1A、VEGF的定位与表达量,利用Real-time PCR检测二者mRNA相对表达量;分析HIF1A与VEGF的表达变化趋势与相关性。结果 P4撤退后,小鼠子宫内膜发生崩解出血,模拟月经发生。免疫组化结果显示,HIF1A蛋白在P4撤退后12、16h,内膜基质蜕膜细胞核中的表达量逐渐升高;HIF1A表达区域与VEGF表达的区域均集中在子宫内膜蜕膜化区域外周,即崩解组织与基底层交界区域。Western blot蛋白定量结果显示,细胞核中HIF1A蛋白表达量在P4撤退后8、12、16h显著增高,VEGF在细胞质中的表达量在P4撤退后0、8、12h较高。HIF1A与VEGF mRNA的表达变化趋势非常相似,即P4撤退后,逐渐升高,在12h达到峰值,随后逐渐下降。结论结果提示月经发生子宫内膜崩解中,HIF1A转录因子功能激活并调节VEGF mRNA的表达。
Objective To study the expression and localization of hypoxia inducible factor (HIF1A) and vascular endothelial growth factor (VEGF) protein and mRNA on the basis of mouse model of menstrual cycle and to explore the possible regulatory mechanism of HIF1A and VEGF in the pathogenesis of menstruation. Methods Mice menstrual model was established. The pseudopregnant mice induced the decidualization of endometrium by peanut oil intrauterine injection and the withdrawal of progesterone (P4) withdrawal from ovary to simulate the occurrence of menstruation. Mouse uteri were collected at 0, 8, 12, 16 and 24 h after P4 retreat. The localization and expression of HIF1A and VEGF were detected by immunohistochemistry and Western blot. Real-time PCR was used to detect the relative expression of both mRNAs. Analysis of HIF1A and VEGF expression trends and correlation. Results P4 withdrawal, the endometrial disintegration occurred in mice to simulate the occurrence of menstruation. The results of immunohistochemistry showed that the expression of HIF1A protein in decidual nucleus of endometrial stroma increased gradually at 12 and 16 h after withdrawal of P4. The expression of HIF1A and VEGF were all concentrated in the periphery of endometrial decidualization, That is, disintegration of the organization and the basal layer junction area. The results of Western blot showed that the expression of HIF1A in nucleus was significantly increased at 8, 12 and 16 h after P4 retreat, and the expression of VEGF in cytoplasm was higher at 0, 8 and 12 h after withdrawal of P4. HIF1A and VEGF mRNA expression trends are very similar, that P4 retreat gradually increased, peaked at 12h, and then gradually decreased. Conclusion The results suggest that during the endometrial disintegration in menstruation, HIF1A transcription factor functionally regulates VEGF mRNA expression.