Dynamic expression of autophagy related genes during endometrial disintegration in a mouse menstrual-like model
Objective:To investigate the expression changes of autophagy genes and its effects during the endometrial disintegration in the mouse menstrual-like model.Methods:A mouse menstrual-like model was established.Mice were executed and bilateral uterine tissues were taken at 0 h,8 h,16 h and 24 h after progesterone withdrawal.The morphology of the endometrium was observed by hematoxylin-eosin(HE)staining.The protein of LC3B and p62 were detected by Western blotting(WB),and real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the dynamic expression of Lc3b,p62,Beclin1,Atg3,Atg5,and Atg7 at the mRNA level during the process of endometrial disintegration in mice.Results:At 0 h,8 h,16 h and 24 h after progesterone withdrawal,the color of the uterus gradually changed from light pink to dark red,and the area of endometrial disintegration and necrosis was gradually enlarged,and the decidualized endometrium was completely disintegrated and necrotic at 24 h.The WB results showed that at 16 hours after progesterone withdrawal,the protein expression level of p62 was significantly increased compared to 0 and 8 hours(P＜0.05),and at 24 hours,the protein expression level of p62 was significantly increased compared to other time points(P＜0.01).The expression level of LC3B-Ⅰ protein showed no significant change from 0 h to 8 h after progesterone withdrawal(P＞0.05),but decreased significantly from 8 h to 24 h(P＜0.05).There was no significant change in the protein expression level of LC3B-Ⅱ during the process of endometrial disintegration after progesterone withdrawal(P＞0.05).The RT-qPCR results showed that there was no significant change in the level of p62 mRNA from 0 h to 16 h after progesterone withdrawal(P＞0.05),while at 24 h,the level of p62 mRNA was significantly reduced compared to 8 h and 16 h(P＜0.05).The level of Lc3b mRNA was significantly upregulated at 16 hours after progesterone withdrawal compared to 0 and 8 hours(P＜0.01),while at 24 hours,the level of Lc3b mRNA was significantly decreased than that at 16 hours after progesterone withdrawal(P＜0.01).The expression of Beclin1 mRNA levels significantly decreased at 8,16,and 24 hours after progesterone withdrawal compared to 0 hours(P＜0.05),and the mRNA levels of Beclin1 significantly decreased at 24 hours after progesterone withdrawal compared to 8 and 16 hours(P＜0.01).After progesterone withdrawal from 0 h to 16 h,Atg3,Atg5 and Atg7 were not significantly changed at the mRNA level,whereas they were all down-regulated at the mRNA level than those at other time points(P＜0.05).Conclusions:After progesterone withdrawal,autophagy levels decreased during endometrial disintegration,suggesting that autophagy is involved in negatively regulating the endometrial disintegration process.
万方数据
维普
