自噬在围着床期小鼠子宫内膜中作用初探

doi:10.12280/gjfckx.20210160

摘要/Abstract

摘要：

目的: 探索围着床期小鼠子宫内膜自噬情况。方法: 选择36只ICR妊娠小鼠随机分为空白组、胚胎着床障碍模型组,每组小鼠18只。模型组采用米非司酮制备,每组每日予以生理盐水灌胃,分别于妊娠第4、5、6天脱颈处死小鼠各6只。观察子宫形态,计算各组平均胚胎着床位点数,检测自噬标志物微管相关轻链蛋白3-Ⅱ（LC3-Ⅱ）,透射电镜观察子宫内膜细胞中自噬小体的表达。结果: 空白组妊娠第4天（pd4）子宫无明显串珠样改变,妊娠第5天（pd5）、妊娠第6天（pd6）子宫串珠样改变明显且分布均匀。模型组pd4、pd5未见明显串珠样改变,pd6可见少量串珠稀疏分布。空白组pd6的胚胎着床位点数较pd4增多,差异有统计学意义（P<0.05）;而pd5的胚胎着床位点数与pd4比较有增多,但差异无统计学意义（P>0.05）。模型组pd5、pd6平均胚胎着床位点数分别低于空白组pd5、pd6（P<0.05,P<0.01）。2组LC3-Ⅱ蛋白水平差异无统计学意义（P>0.05）。透射电镜下空白组小鼠子宫内膜细胞完整,pd5自噬小体数量较pd4减少,pd6又增多。模型组部分细胞损伤、坏死,pd4自噬小体表达较空白组多,亦较模型组pd5、pd6多。结论: 自噬参与并调控胚胎着床过程,自噬异常可能破坏子宫内膜细胞活性和功能,影响胚胎着床。

关键词: 自噬, 围着床期, 胚胎植入, 小鼠,近交ICR, 子宫内膜, 印迹法, 蛋白质

Abstract:

Objective:To observe autophagy in mouse endometrium during peri-implantation period.Methods: ICR pregnant mice (n=36) were randomly divided into blank group and embryo implantation disorder model group (each n=18). The model group was prepared with mifepristone. Mice in each group were given normal saline and 6 mice were killed by neck dislocation respectively on the 4th, 5th, 6th day of gestation. The appearance of uterus was observed. The average number of implantation sites was calculated. The protein expression of microtubule associated protein light chain 3 (LC3-Ⅱ) was detected, and autophagosomes in endometrial cells were observed by transmission electron microscope.Results: In control group, there were no obvious bead-like changes in the uterus on the 4th day of gestation(pd4), but that were obvious and evenly distributed on the 5th day of gestation (pd5) and the 6th day of gestation(pd6). There was no obvious bead-like changes in the uterus on pd4 and pd5 in model group, but few and sparsed bead-like changes on pd6. The average number of embryo implantation sites on pd5 and pd6 in control group was higher than that on pd4, and that on pd6 was significantly different compared with that on pd4 (P<0.05). The average number of embryo implantation sites on pd5 and pd6 in model group respectively decreased compared with control group (P<0.05, P<0.01). Both control group and model group showed that the protein expression of LC3-Ⅱ was not significantly different (P>0.05). Under the transmission electron microscope, endometrial cells of control group were intact. The number of autophagosomes in control group on pd5 was decreased, and increased on pd6. Some endometrial cells in model group were necrosis. There were more autophagosomes in model group on pd4 than that in control group, model group on pd5 and pd6.Conclusions: Autophagy participates in and regulates the process of embryo implantation. Abnormal autophagy may destroy the activity and function of endometrial cells, which is harmful to embryo implantation.

Key words: Autophagy, Peri-implantation period, Embryo implantation, Mice,inbred ICR, Endometrium, Blotting, Western

申萌萌, 刘雁峰, 马小娜, 贾静, 王峥, 朱庆文. 自噬在围着床期小鼠子宫内膜中作用初探[J]. 国际妇产科学杂志, 2021, 48(6): 666-669.

SHEN Meng-meng, LIU Yan-feng, MA Xiao-na, JIA Jing, WANG Zheng, ZHU Qing-wen. Preliminary Study on the Role of Autophagy in Mouse Endometrium during Peri-Implantation[J]. Journal of International Obstetrics and Gynecology, 2021, 48(6): 666-669.

图/表 4

图1 各组小鼠子宫形态注：A 空白组pd4;B 空白组pd5;C 空白组pd6;D 模型组pd4;E 模型组pd5;F 模型组pd6。

表1 各组小鼠妊娠第4、5、6天胚胎着床位点数 [M（P25,P75）]

组别	n	pd4	pd5	pd6	χ²	P
空白组	6	4（0,14.00）	10.50（8.75,12.00）	15.0（11.00,16.25）*	6.38	0.04
模型组	6	0（0,2.00）	3.50（1.50,7.00）	4.5（0.75,6.50）	4.44	1.09
T或Z		-1.074	4.93	6.19
P		0.28	0.01	0.00

图2 围着床期小鼠子宫内膜中LC3-Ⅱ蛋白的表达（n=3）注：与空白组pd4相比,*P<0.05。

图3 围着床期小鼠子宫内膜细胞中自噬小体的表达（×5 000）注：A 空白组pd4;B 空白组pd5;C 空白组pd6;D 模型组pd4;E 模型组pd5;F 模型组pd6。图中白色箭头所指为自噬小体。

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