生理性孕酮撤退过程中小鼠子宫角组织蛋白激酶B、糖原合成激酶3β的表达观察

摘要

目的 观察生理性孕酮撤退小鼠子宫角组织中蛋白激酶 B(Akt)及糖原合成激酶3β(GSK3β)的表达变化.方法 90只未交配健康成年雌性C57BL/6J小鼠,制作生理性孕酮撤退模型,分别在孕酮撤退0、8、12、16、20、24、32、40、48 h(48 h为小鼠生理性孕酮撤退周期)时处死10只小鼠,收集子宫角组织.采用real-time PCR法检测孕酮撤退各时点小鼠子宫角组织Akt、GSK3β mRNA,采用Western blotting法检测孕酮撤退各时点小鼠子宫角组织总蛋白激酶 B(t-Akt)、磷酸化蛋白激酶 B(Akt)及3βp-Akt及磷酸化糖原合成激酶(p-GSK3β).结果 随孕酮撤退时间延长,Akt mRNA及蛋白相对表达量逐渐降低,孕酮撤退20 h时达到最低值,后随孕酮撤退时间逐渐延长,Akt mRNA及蛋白相对表达量逐渐升高.孕酮撤退8 h 时GSK mRNA及蛋白相对表达量高于孕酮撤退0 h时(P<0.05);随孕酮撤退时间延长,GSK mRNA及蛋白相对表达量逐渐降低,孕酮撤退24 h时达到最低值,后随孕酮撤退时间逐渐延长, GSK mRNA及蛋白相对表达量逐渐增加.孕酮撤退20 h小鼠子宫角组织 Akt 、GSK3β mRNA表达量低于孕酮撤退后0、48 h(P均<0.05).与孕酮撤退0 h比较,孕酮撤退20 h时小鼠子宫角组织p-Akt、p-GSK3β的相对表达量均降低(P均<0.05).与孕酮撤退8 h比较,孕酮撤退20 h时小鼠子宫角组织p-GSK3β的相对表达量降低(P<0.05).与孕酮撤退20 h比较,孕酮撤退48 h时小鼠子宫角组织p-Akt、p-GSK3β的相对表达量均升高(P均<0.05).结论 生理性孕酮撤退小鼠子宫角组织中存在Akt表达随时间下降后上调,GSK3β表达随时间上调后下降后再次上调. Akt/GSK-3β信号通路可能在鼠孕酮撤退过程中起重要作用.%Objective To observe the expression of protein kinase B (Akt) and glycogen synthase kinase 3β (GSK3β) in the uterine tissues of mouse models in the process of physiological progesterone withdrawal.Methods We selected 90 non-mating healthy adult female C57BL/6J mice to make the physiological progesterone withdrawal models.Ten mice were killed at 0, 8, 12, 16, 20, 24, 32, 40, and 48 h (The physiological progesterone withdrawal cycle in mice is 48 h) to collect uterine horns.The gene expression levels of Akt and GSK3β were detected by real-time PCR.The protein expression levels of total Akt, phosphorylated AKT and phosphorylated GSK-3β were detected by Western blotting.Results Over the time of progesterone withdrawal, the expression of Akt mRNA and protein gradually decreased (reached the minimum at 20 h) and then gradually increased.The expression of GSK3β mRNA and protein at 8 h was higher than that at 0 h (P<0.05).Over the time of progesterone withdrawal, the expression of GSK mRNA and protein decreased (reached the minimum at 24 h) and then gradually increased.The expression of Akt and GSK3β mRNA at 20 h was lower than that at 0 and 48 h (both P<0.05).The expression of p-Akt and p-GSK-3β at 20 h decreased as compared with that at 0 h (both P<0.05).The expression of p-GSK-3β protein in mouse uterine tissues at 20 h decreased as compared with that at 8 h (P<0.05).The expression of p-Akt and p-GSK3β protein at 48 h increased as compared with that at 20 h (all P<0.05).Conclusions Over the time of progesterone withdrawal, the expression of Akt gradually decreases and then increases in the uterine tissues of mouse models.Meanwhile, the expression of GSK3β first increases, then decreases, and finally increases again.Akt/GSK3β signal pathway may play an important role in the process of progesterone withdrawal.