miR-133a_R+1调控ALDH5A1的表达对奶牛胎衣不下的影响  

The Effect of miR-133a_R+1 Regulation to ALDH5A1 Expression on Retained Placenta in Cows(Bos taurus)

作　　者：汪玥 吕琛 姚丹 赵兴绪[1] 张勇[1] WANG Yue;LYU Chen;YAO Dan;ZHAO Xing-Xu;ZHANG Yong(Gansu Key Laboratory of Animal Generational Physiology and Reproductive Regulation/College of Veterinary Medicine,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院/甘肃省动物生殖生理及繁殖调控重点实验室,兰州730070

出　　处：《农业生物技术学报》2024年第11期2564-2571,共8页Journal of Agricultural Biotechnology

基　　金：奶牛围产期疾病早期预警技术研发与示范(21JR7RA858);甘肃省动物生殖生理及繁殖调控重点实验室项目(20JR10RA563);牛羊围产期群发普通病防控技术研发与示范甘肃省引导科技创新专项资金(GSCXZX-2019)。

摘　　要：Micro RNA(miRNA)参与哺乳动物许多生理活动,其中miR-133a在胎盘和子宫生长发育过程中具有重要的调控作用。通过测序及靶基因预测软件发现miR-133a_R+1与ALDH5A1在胎衣不下奶牛(Bos taurus)胎盘组织中差异表达且可能存在靶向调控关系。本研究以正常排出及胎衣不下奶牛胎儿胎盘组织和健康奶牛子宫内膜上皮细胞(bovine endometrial epithelial cells,BEND)为实验对象,苏木精-伊红染色(hematoxylin-eosin,HE)观察胎儿胎盘组织的病理变化,实时定量PCR(quantitative real-time PCR,qRT-PCR)检测miR-133a_R+1与乙醛脱氢酶5家族成员A1(aldehyde dehydrogenase 5 family member A1,ALDH5A1)在胎儿胎盘组织中的表达情况,采用免疫组织化学法(immunohistochemistry,IHC)检测ALDH5A1蛋白在胎儿胎盘组织中的定位,选取转染效率最佳条件100 nmol/L、12 h构建过表达miR-133a_R+1的BEND细胞模型,通过qRT-PCR与Western blot检测细胞中ALDH5A1基因及其蛋白表达水平。结果显示,RP胎儿胎盘绒毛短而稀疏,零散破碎。RP胎儿胎盘组织中miR-133a_R+1基因的表达量极显著上调(P<0.01),ALDH5A1在结缔组织、细胞滋养层中表达且表达量极显著下调(P<0.01),过表达miR-133a_R+1的BEND细胞中ALDH5A1基因及蛋白表达水平均极显著下调(P<0.01)。实验结果表明,miR-133a_R+1通过靶向调控ALDH5A1的表达影响奶牛RP,为今后miRNA在奶牛RP中的作用机制研究提供了理论基础。Micro RNA(miRNA)is involved in numerous physiological activities in mammals,with miR133a playing a pivotal regulatory role in placental and uterine growth and development.miR-133a_R+1 and ALDH5A1 are differentially expressed in placental tissues of cows(Bos taurus)with retained placenta and may have a potential targeting regulatory relationship through sequencing and target gene prediction software.In this study,normal fetal placental tissue,RP placental tissue,and bovine endometrial epithelial cells(BEND)were used as experimental materials.Hematoxylin eosin(HE)staining was used to observe the pathological changes of placental tissue,and qRT-PCR was used to detect expression of miR-133a_R+1 and aldehyde dehydrogenase 5 family member A1(ALDH5A1)in placental tissue.Immunohistochemistry(IHC)was used to detect the distribution of ALDH5A1 protein in placental tissue.Optimal transfection conditions was 100 nmol/L 133a_R+1 mimic for 12 h to establish a BEND cell model overexpressing miR-133a_R+1.qRT-PCR and Western blot were used to detect the expression of ALDH5A1 gene and its protein in the cells.The results showed that RP placental tissue villi were short,sparse,and fragmented.The expression level of miR-133a_R+1 mRNA in RP placenta tissue was extremely significantly up-regulated(P<0.01),ALDH5A1 was expressed in connective tissue and cytotrophoblast,and the expression level was extremely significantly down-regulated(P<0.01).The expression levels of ALDH5A1 gene and its protein in BEND cells transfected with miR133a_R+1 mimic were extremely significantly downregulated(P<0.01).In summary,miR-133a_R+1 affected cow RP by targeting the expression of ALDH5A1 gene and its protein.This study provides corresponding theoretical basis for further exploring the mechanism of miRNA action on dairy cow RP.

关 键 词：奶牛 胎衣不下 胎盘 miR-133a_R+1 乙醛脱氢酶5家族成员A1(ALDH5A1) 

分 类 号：S823.91[农业科学—畜牧学]