小鼠子宫内膜中AMPK蛋白及mRNA表达对胚胎着床调控作用的实验研究  

Experimental Study on the Regulation of AMPK Protein and mRNA Expression in Mouse Endometrium on Embryo Implantation

作　　者：孟靠 马娟 MENG Kao;MA Juan(Department of Reproductive Medicine,Suzhou Hospital Affiliated to Anhui Medical University/Suzhou Municipal Hospital,Anhui Suzhou 234000,China)

机构地区：[1]安徽医科大学附属宿州医院/宿州市立医院生殖医学科,安徽宿州234000

出　　处：《现代检验医学杂志》2024年第6期43-47,共5页Journal of Modern Laboratory Medicine

基　　金：安徽省卫生健康委员会科研项目(2021538AJH)。

摘　　要：目的探究妊娠小鼠子宫内膜中AMP依赖的蛋白激酶(AMP-activated protein kinase,AMPK)表达对胚胎着床调控作用的实验研究。方法使用ICR小鼠建立1~8天正常妊娠小鼠、1~5天假妊娠小鼠模型、阴性对照(NC)组和AMPK抑制剂(Dorsomorphin)组。免疫组织化学染色检测小鼠子宫内膜中AMPK蛋白的表达位置,逆转录实时定量聚合酶链式反应(RT-qPCR)检测子宫内膜中AMPK及子宫着床处相关指标黏蛋白1(Muc1)、同源框蛋白A10(HoxA10)和心脏神经嵴衍生物表达蛋白2(Hand2)的mRNA水平,Western blotting检测子宫内膜中AMPK蛋白水平,统计妊娠小鼠胚泡数量。结果妊娠第1~5天,AMPK蛋白及mRNA表达逐渐增加(F=683.50,678.20,均P<0.05),且主要表达于子宫内膜管腔细胞和腺上皮细胞。与妊娠第5天相比,妊娠第6天小鼠子宫AMPK蛋白及mRNA表达减少(t=25.98,68.41,均P<0.05),且主要表达于初级蜕膜化区(PDZ)和次级蜕膜化区(SDZ),妊娠第6~8天,AMPK蛋白及mRNA在子宫内膜中表达逐渐增加(t=86.00,54.05;29.96,127.60,均P<0.05)。与假妊娠第1天比较,假妊娠第2~5天,AMPK mRNA及蛋白的表达减少(t=38.97,30.05,33.00,13.98;22.71,26.90,53.81,65.00,均P<0.05),且主要表达于腺体和管腔上皮细胞。与NC组小鼠(18.20±3.19)相比,Dorsomorphin组小鼠(9.40±2.41)平均胚泡数量减少(t=8.73,P<0.05),子宫着床处Muc1 mRNA(5.97±0.32 vs 1.10±0.15)表达增加,HoxA10(0.20±0.10vs 1.03±0.06)及Hand2(0.23±0.15 vs 0.97±0.12)mRNA表达减少(t=32.15,3.46,2.65,均P<0.05)。结论小鼠子宫内膜组织中AMPK在胚胎着床中发挥重要作用,抑制该蛋白表达将不利于胚胎着床。Objective To investigate the regulatory effect of AMP-activated protein kinase(AMPK)protein and mRNA expression in endometrium of pregnant mice on embryo implantation.Methods ICR mice were used to establish 1~8 days normal pregnancy mice and 1~5 days pseudopregnancy mice models,which were divided into negative control(NC)group and AMPK inhibitor(Dorsomorphin)group.Immunohistochemical staining was used to detect the expression position of AMPK protein in the endometrium of mice.Reverse transcripatase quantitative polymerase chain reaction(RT-qPCR)was used to detect the AMPK in the endometrium of mice,and the related indicators of implantation in the uterus including the mRNA levels of AMPK,mucin1(Muc1),homeobox A10(Hoxa10)and cardiac neural ridge derivative expression protein 2(Hand2).Western blotting was applied to detect AMPK protein levels in endometrium and count the number of embryonic blastocysts in pregnant mice.Results The expression of AMPK protein and mRNA were gradually increased from day 1st to day 5th of pregnancy(F=683.50,678.20,all P<0.05),and mainly expressed in endometrial lumen cells and glandular epithelial cells.Compared with day 5th of gestation,the expressions of AMPK protein and mRNA in uterus of micedecroused on the 6th day of gestation(t=25.98,68.41,all P<0.05),and the expression was mainly in the primary decidualizing zone(PDZ)and the secondary decidualizing zone(SDZ).The expressions of AMPK protein and mRNA in endometrium were gradually increased on the 6th to 8th day of pregnancy(t=86.00,54.05;29.96,127.60,all P<0.05).Compared with 1st day of pseudogestation,the expression of AMPK mRNA and protein decreased on days 2~5 of pseudogestation(t=38.97,30.45,33.00,13.98;22.71,26.90,53.81,65.00,all P<0.05),,and mainly expressed in gland and luminal opithelial cells.Compared with NC group,Muc1 mRNA(5.97±0.32 vs 1.10±0.15)expression increased,(18.20±3.19),Dorsomarphin group(9.40±2.41)reuced the mean number of blastoblasts(t=8.73,P<0.05),HoxA10(0.20±0.10 vs 1.03±0.06)and Hand2(0.23±0.15 vs

关 键 词：胚胎着床 子宫内膜 AMP依赖的蛋白激酶 早孕小鼠 

分 类 号：R-332[医药卫生]