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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。
This study analyzed the expression pattern of pyruvate kinase M2 (PKM2) gene in the endometrium of early pregnant mice. Through establishing the normal pregnancy mouse model, the endometrial tissues of pregnant D1, D4, D5, D6, D7 mice and the implantation site of pregnant D5 mice and the endometrial tissue adjacent to the implantation were collected. The model of pseudopregnancy mice was established and the endometrial tissues of pseudopregnant PD1, PD4, PD5, PD6 and PD7 mice were collected. The expression of PKM2 mRNA and protein were detected by Real-time PCR and Western blot. The distribution of PKM2 protein at the site of implantation at pregnancy and the implantation of endometrium were detected by immunohistochemistry. The results showed that the expression of PKM2 m RNA in normal pregnant mice endometrium was significantly increased from the first pregnancy D5, reached the peak in pregnancy D6, a slight decrease in pregnancy D7, pregnant D6, pregnant D7 compared with the pregnancy D1 significant difference . PKM2 protein began to appear significantly increased from pregnancy D6, a slight decrease in pregnancy D7, pregnancy D6, pregnancy D7 and pregnancy D1 compared to significant differences. The level of PKM2 m RNA in endometrium of pseudopregnant mice significantly increased from PD6, PD7 and PD6 levels were comparable, PD6, PD7 and PD1 were significantly different. There was no significant difference in PKM2 protein levels between the two groups. Pregnant D5 mouse endometrial tissue, PKM2 m RNA and protein levels showed a significantly higher implantation point than the implantation side trend. The study initially revealed the expression of PKM2 gene in the endometrium of early pregnant mice and provided important clues for further exploring the role of PKM2 in maintaining the normal endometrium function of early pregnant mice.