oar-miR-103和oar-miR-107通过靶向IL-1β基因区域调节其表达  

oar-miR-103 and oar-miR-107 Regulate Expressions of IL-1βGene by Targeting Their Regions

作　　者：高之煜 顾兰英 范文雨 曹鑫艳 陈创夫[1] 肖非 易继海 盛金良[1] 张彦兵[1,2] 孙延鸣 GAO Zhi-yu;GU Lan-ying;FAN Wen-yu;CAO Xin-yan;CHEN Chuang-fu;XIAO Fei;YI Ji-hai;SHENG Jin-liang;ZHANG Yan-bing;SUN Yan-ming(College of Animal Science and Technology,Shihezi University,Shihezi,Xinjiang,832003,China;Xinjiang Tycoon Group Co.,Ltd.,Changji,Xinjiang,831203,China)

机构地区：[1]石河子大学动物科技学院,新疆石河子832003 [2]新疆泰昆集团股份有限公司,新疆昌吉831203

出　　处：《动物医学进展》2025年第6期31-35,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(32360901,32060133);兵团指导性科技计划项目(2023ZD042);新疆维吾尔自治区青年科学基金项目(2022D01B197);石河子大学高层次人才科研启动项目(RCZK202043)。

摘　　要：为确定oar-miR-103和oar-miR-107靶向IL-1β基因的区域及结合能力,验证绵羊oar-miR-103和oar-miR-107对绵羊IL-1β表达的调控作用,用miRanda软件分析oar-miR-103、oar-miR-107靶向IL-1β基因的具体区域和结合能力;将IL-1β基因插入PGL3-promoter荧光素酶报告基因载体XbaⅠ酶切位点,分别将miR-NC、miR-103和miR-107模拟物与荧光素酶质粒共转染,通过双荧光素酶报告试验分析oar-miR-103和oar-miR-107与IL-1β基因区域靶向关系;分离原代绵羊肺泡巨噬细胞,miR-NC、miR-103和miR-107模拟物分别转染原代肺泡巨噬细胞,转染36 h后,通过实时荧光定量PCR(RT-qPCR)检测各组细胞中IL-1β表达水平。结果显示,oar-miR-103和oar-miR-107分别靶向IL-1β基因2个、3个区域;成功构建了荧光素酶报告载体pGL3-promoter-IL-1β;双荧光素酶试验显示,与miR-NC组相比,oar-miR-103和oar-miR-107两组均使pGL3-promoter-IL-1β活性显著下降(P<0.01);与miR-NC组相比,oar-miR-107显著抑制了巨噬细胞IL-1βmRNA转录表达(P<0.05)。说明oar-miR-103和oar-miR-107可通过靶向IL-1β基因区域调节其表达,为解析IL-1β表达机理积累了资料。To investigate whether sheep microRNAs(miRNAs)target the IL-1βgene,and to initially determine the specific region of the miRNA that targets its gene,and to verify the regulatory effect of miRNAs on IL-1βexpression,taking sheep miRNAs as the research object,the miRanda software was used to analyze the specific regions and binding ability of miRNAs targeting the IL-1βgene.The sheep IL-1βgene was connected to a luciferase reporter gene vector,and the targeting effect of oar-miR-103 and oar-miR-107 on the IL-1βgene region was verified through a luciferase reporter gene experiment.Primary sheep alveolar macrophages were isolated and overexpressed with oar-miR-103 and oar-miR-107 to analyze the IL-1βtranscript expression level.The results showed that oar-miR-103 and oar-miR-107 targeted two and three regions of the IL-1βgene,respectively.A luciferase reporter vector pGL3-promoter-IL-1βwas successfully constructed,and the dual luciferase assay was used to detect the targeting effect of oar-miR-103 and oar-miR-107 on the IL-1βgene region.It was found that compared with the miR-NC group,both oar-miR-103 and oar-miR-107 groups significantly decreased the activity of pGL3-promoter-IL-1β.Cell experiments demonstrated that oar-miR-107 significantly inhibited the transcription and expression of IL-1βmRNA.The results demonstrated that oar-miR-103 and oar-miR-107 have the ability to target and inhibit the expression of IL-1βgene,providing basic information for further exploring the regulation of miRNAs on IL-1βexpression.

关 键 词：白介素1Β MICRORNAS MIRANDA 荧光素酶报告基因 

分 类 号：S852.2[农业科学—基础兽医学]