microRNA-654调控巨噬细胞游走抑制因子抗东方泰勒虫感染长角血蜱的作用  

Regulation of microRNA-654 in MIF of Haemaphysalis longicornis for anti-infection of Theileria orientalis

作　　者：罗金 刘文阁[2] 任巧云[2] 严若峰[1] 宋小凯[1] 刘光远[2] 李祥瑞[1] LUO Jin;LIU Wen-ge;REN Qiao-yun;YAN Ruo-feng;SONG Xiao-kai;LIU Guang-yuan;LI Xiang-rui(MOE Joint International Research Laboratory of Animal Health and Food Safety/College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;State Key Laboratory of Veterinary Etiological Biology/Key Laboratory of Veterinary Parasitology of Gansu Province/Key Open Laboratory of Herbivores Animal Epidemics,Ministry of Agriculture/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)

机构地区：[1]南京农业大学动物医学院教育部动物健康与食品安全国际合作联合实验室,江苏南京210095 [2]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室农业部草食动物疫病重点开放实验室,甘肃兰州730046

出　　处：《中国兽医科学》2022年第8期1023-1030,共8页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2019YFC1200502,2019YFC1200500,2017YFD0501206);国家寄生虫资源库项目(NPRC-2019-194-30);国家肉牛牦牛产业技术体系项目(CARS-37);中国农业科学院科技创新工程项目(Y2019YJ07-04,Y2018PT76)。

摘　　要：为了解microRNA(mi RNA)调控巨噬细胞游走抑制因子(MIF)抗东方泰勒虫感染长角血蜱的作用,利用PCR扩增获得MIF基因的完整ORF,并进行体外原核表达,制备抗原免疫家兔,验证MIF重组蛋白对长角血蜱发育的影响。另外,预测可能调控MIF基因的mi RNA分子,用双荧光素酶报告系统分析mi RNA与靶标基因之间的调控关系,并解析此作用下靶标基因引起的抗病原作用。结果显示,MIF基因是一个包含348 bp核酸,编码116个氨基酸的片段。动物免疫试验证实,MIF+GST重组蛋白对蜱发育的各项生理指标影响较小。并且MIF受mi R-654的负调控,其调控效率达到60%。在蜱体中,mi RNA抑制后的MIF表达对东方泰勒虫感染不同发育阶段的长角血蜱有一定的识别和清除作用,可降低蜱体中约55%的虫体。尽管MIF对蜱的发育没有明显影响,但是在蜱抗病原感染过程中扮演着重要的角色。mi RNA作为功能基因的重要调控因子,在MIF识别和清除病原方面发挥着重要作用。为进一步蜱传疾病的防控提供了新的策略。To investigate the effect of macrophage migration inhibitory factor(MIF) mediated by micro RNA on Haemaphysalis longicornis infected by Theileria orientalis,the maximum ORF of MIF gene was obtained by PCR and expressed in vitro,and immune rabbits were prepared.In addition,it was predicted for micro RNA(mi RNA) may regulate MIF,and the regulatory relationship was analyzed by dual luciferase reporter system.The against pathogens effect of target genes was analyzed under the micro RNA regulatory mechanism.The regulation of micro RNA was used to analyze the anti-pathogenic effect of the target gene.The MIF gene was a 348 bp fragment,and encoding 116 amino acids.Animal experiments showed that the MIF+GST recombinant protein had little effect on the physiological indexes of the development of ticks.Target gene prediction indicated that mi R-654 could negatively regulate MIF expression,with 60%.The MIF protein had a function for recognize the T.orientalis infected different developmental stages of H.longicornis by inhibiting mi RNA for MIF expression,and the clearance rate was 55%.Although MIF has no significant effect on ticks development,but it plays an important role in ticks against pathogens infection.Micro RNA,as an important regulator factor for functional genes,plays an important role in MIF recognition and clearance of pathogens.And it provides a new strategy for further prevention and control of tick-borne diseases.

关 键 词：巨噬细胞游走抑制因子 MICRORNA 长角血蜱 东方泰勒虫 抗感染 

分 类 号：S852.723[农业科学—基础兽医学]