早孕小鼠子宫内膜M2型丙酮酸激酶基因表达的研究

The Study of the Expression of Pyruvate Kinase M2 Gene in the Endometrium Mouse during the Early Stage of Pregnant

该研究分析了M2型丙酮酸激酶(pyruvate kinase M2,PKM2)基因在早孕小鼠子宫内膜的表达规律。通过建立正常妊娠小鼠模型,收集孕D1、D4、D5、D6、D7小鼠子宫内膜组织及孕D5小鼠着床点及着床旁子宫内膜组织。构建假孕小鼠模型,收集假孕PD1、PD4、PD5、PD6和PD7小鼠子宫内膜组织。用Real-time PCR和Western blot方法检测PKM2 m RNA和蛋白质表达水平;免疫组织化学方法检测PKM2蛋白质在孕D5着床点与着床旁子宫内膜的分布。研究结果显示,在正常妊娠小鼠子宫内膜,PKM2 m RNA表达从孕D5开始出现明显升高,孕D6达高峰,孕D7略有下降,孕D6、孕D7与孕D1相比有明显差异。PKM2蛋白质从孕D6开始出现明显升高,孕D7略有下降,孕D6、孕D7与孕D1相比有明显差异。假孕小鼠子宫内膜PKM2 m RNA水平从PD6开始有明显升高,PD7与PD6水平相当,PD6、PD7与PD1相比有明显差异。PKM2蛋白质水平每两组间无明显差异。孕D5小鼠子宫内膜组织中,PKM2 m RNA及蛋白质水平均呈现着床点明显高于着床旁趋势。该研究初步揭示了PKM2基因在早孕小鼠子宫内膜表达规律,为深入探讨PKM2在维持早孕小鼠子宫内膜正常功能的机制上的作用提供了重要线索。

The expression rules of the type of pyruvate kinase M2（PKM2） gene in the endometrium of mouse during the early stage of pregnant were analyzed in this study. By establishing the pregnant model, we collected the endometrium of pregnant mice on D1, D4, D5, D6, D7 and implantation site and inter-implantationsite on D5. By establishing the pseudopregnant model, we collected the endometrium of pseudopregnant mice on PD1, PD4, PD5, PD6, PD7. Real-time PCR and Western blot were employed to detect the expressions of PKM2. Immunohistochemistry was used to detect the distribution of PKM2 protein in the endometrium of mice at implantation site and inter implantation site on D5. The results revealed that the level of PKM2 m RNA in the endometrium of pregnant miceraised from D5, and reached the peak on D6, declined slightly on D7. The level of PKM2 protein in the endometrium of mice pregnant raised from D6, declined slightly on D7. The level of PKM2 protein on D6 and D7 had obvious differences compared with D1. In pseudopregnant model, the level of PKM2 mR NA in the endometrium of mice raised from PD6, consistented with PD6 and PD7. The level of PKM2 mR NA of PD6 and PD7 have obvious differences compared with PD1. There was no significant difference of PKM2 protein expression in pseudopregnant model. The level of PKM2 mR NA and protein in the endometrium of mice at implantation site was obviously higher than inter-implantation site on D5. Our results preliminarily revealed the expression rules of PKM2 gene in the endometrium, also provided an important clue to explore the mechanism how the PKM2 gene maintained the normal functions in the endometrium of mouse during the early stage of pregnant.