miR-1468对延边黄牛垂体细胞生长激素分泌影响的研究  

The mechanism of the effect of miR-1468 on GH secretion in the pituitary cells of Yanbian yellow cattle

作　　者：相思宇 季久秀 张睿[1] 金太花 陈莹莹[1] 娄安钢[1] 崔长艳 于龙政[1] 关立增[1,2] XIANG Siyu;JI Jiuxiu;ZHANG Rui;JIN Taihua;CHEN Yingying;LOU Angang;CUI Changyan;YU Longzheng;GUAN Lizeng(College of Agriculture,Yanbian University,Yanji 133002,China;College of Agriculture and Forestry Sciences,Linyi University,Linyi 276005,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]临沂大学农林科学学院,山东临沂276005

出　　处：《畜牧与兽医》2020年第9期13-19,共7页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31660614)。

摘　　要：为研究血液外胞体中miRNA对延边黄牛垂体细胞中生长激素(GH)分泌的影响,本研究选择在延边黄牛和韩延牛血液外胞体中显著差异表达的miR-1468,利用实时定量PCR(qPCR)和Western blot技术,研究了miR-1468对延边黄牛垂体细胞中GH分泌水平的影响及miR-1468与靶基因之间的调控机制。结果显示,miR-1468水平的增加能显著抑制延边黄牛垂体细胞中GH mRNA和蛋白的表达(P<0.01),反之降低miR-1468水平能显著提高GH mRNA(P<0.05)和蛋白(P<0.01)的表达。生物信息学分析和双荧光素酶报告基因系统结果显示,miR-1468靶向了环磷腺苷效应元件结合蛋白1(CREB1)的3′UTR;miR-1468水平的增加能够显著抑制CREB1 mRNA和蛋白的表达(P<0.01),反之降低miR-1468水平能显著提高CREB1 mRNA和蛋白的表达(P<0.01)。本研究表明,miR-1468可通过调节CREB1基因的表达而调控垂体细胞GH的分泌,该结果将为研究外胞体miRNA调控动物生长发育机制提供理论依据。In order to study the effect of miRNAs on GH secretion in the pituitary cells of Yanbian yellow cattle,the differently expressed miR-1468 in the blood of Yanbian yellow cattle and Han-Yan cattle was obtained to determine the effect of miR-1468 on GH secretion in pituitary cells of Yanbian yellow cattle and the regulation mechanism between miR-1468 and target genes using Real-time quantitative PCR(qPCR)and Western blot.The results qPCR and Western blot showed that the expressions of GH mRNA and protein in the pituitary cells of Yanbian yellow cattle were significantly decreased by increasing the level of miR-1468(P<0.01),whereas the decrease of the level of miR-1468 enhanced the expressions of GH mRNA(P<0.05)and protein(P<0.01).The results of bioinformatics analysis and double luciferase reporter gene system validation showed that miR-1468 targeted 3′UTR of the cAMP responsive element binding protein 1(CREB1).The increase of the level of miR-1468 significantly inhibited the expressions of the CREB1 mRNA and protein(P<0.01),but the decrease of the level of miR-1468 significantly increase the expressions of the CREB1 mRNA and protein(P<0.01).This study proved that miR-1468 regulated GH secretion in pituitary cells by regulating the expression of the CREB1 gene,which provided a theoretical basis for research on the mechanism of growth and development of animals regulated by exosomal miRNAs.

关 键 词：miR-1468 延边黄牛 垂体细胞 生长激素 环磷腺苷效应元件结合蛋白1(CREB1) 

分 类 号：S823.8[农业科学—畜牧学]