miRNAs-MUC 13调控体系中与仔猪腹泻有关的基因与通路  

Genes and pathways related to piglet diarrhea in miRNAs-MUC13 regulation system

作　　者：路伏增[1] 华卫东[1] 褚晓红[1] 戴丽荷[1] 陈晓宇[1] 章利丰[1] 王志刚[1] 徐如海[1] LU Fuzeng;HUA Weidong;CHU Xiaohong;DAI Lihe;CHEN Xiaoyu;ZHANG Lifeng;WANG Zhigang;XU Ruhai(Institute of Animal Husbandry and Veterinary Medicine,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)

机构地区：[1]浙江省农业科学院畜牧兽医研究所,浙江杭州310021

出　　处：《浙江农业学报》2023年第12期2785-2793,共9页Acta Agriculturae Zhejiangensis

基　　金：浙江省农业新品种选育重大科技专项(2021C02068-4);国家重点研发计划子任务(2021YFD2000801);浙江省2021—2023年畜牧产业技术项目(2022R22S60D02);浙江省基础公益研究计划(LGN22C170009)。

摘　　要：为研究microRNAs-Mucin 13(miRNAs-MUC 13)调控体系中与仔猪腹泻相关的分子调控机制,对miRNAs-MUC 13调控体系中与仔猪腹泻有关的基因与通路进行了发掘与验证。用NCBI的blastn工具比对了小鼠MUC 13基因和猪基因库中的核酸序列,发现两者的相似性在74%~87%,同源性较高。以小鼠为模型,选择miRDB、mirTarBase、TargetScan数据库预测调控MUC 13基因的miRNA序列,通过StarBase数据库预测这些miRNA序列的靶基因,得到36个靶基因交集。用Cytoscape软件围绕MUC 13和这36个靶基因构建仔猪腹泻基因互作网络,该网络包含19个筛选到的靶基因,基于MCOMD算法分析获得4个分子复合物集团。通过DAVID平台对这4个分子复合物进行基因和通路富集分析,结果发现,靶基因SRF和STAG 2及其参与的有关通路与仔猪腹泻相关。以大肠埃希菌200和仔猪肠上皮细胞系(IPEC-1)作为试验材料,通过体外细胞试验设计仔猪腹泻模型进行验证,实时荧光定量PCR(qRT-PCR)检测结果显示,试验组MUC 13、SRF、IL-6、MAPK、STAG 2、NF-κB、TLR 7等基因的表达量均显著(P<0.05)高于对照组,由此推测,SRF与STAG 2基因在miRNAs-MUC 13调控体系中分别通过MAPK、IL6、Toll-like受体信号通路参与仔猪腹泻调控。围绕miRNAs-MUC 13调控体系发掘了4个分子复合物集团和2个核心基因(SRF和STAG 2)并进行了试验验证,这为靶向治疗仔猪腹泻提供了理论参考和科学依据。In order to discover the molecular regulatory mechanism related to piglet diarrhea,the genes and pathways related to piglet diarrhea in the microRNAs-Mucin 13(miRNAs-MUC 13)regulatory system were explored and verified.NCBI blastn tool was used to compare the nucleic acid sequences of mouse MUC 13 gene and pig gene bank,and found that the similarity between them was 74%-87%,with high homology.Taking the mouse as a model,the miRDB,mirTarBase and TargetScan databases were selected to predict the miRNA sequence that regulated the MUC 13 gene.The target genes of these miRNA sequences were predicted through the StarBase database,and 36 target gene intersections were obtained.The interaction network of piglet diarrhea genes was built around MUC 13 and these 36 target genes by using Cytoscape software.The network contains 19 screened target genes,and 4 molecular complex groups were obtained based on MCOMD algorithm analysis.The gene and pathway enrichment analysis of these 4 molecular complexes were carried out through DAVID platform.The results showed that the target genes SRF and STAG 2 and their involved pathways were related to piglet diarrhea.Using Escherichia coli strain 200 and piglet intestinal epithelial cell line(IPEC-1)as the test materials,the piglet diarrhea model was designed and verified by the in vitro cell test.The real-time fluorescence quantitative PCR(qRT-PCR)results showed that the relative expressions of MUC13,SRF,IL-6,MAPK,STAG2,NF-κB,TLR 7 in the test group were significantly higher than those of the control group(P<0.05),so it was speculated that SRF and STAG 2 genes were involved in the regulation of piglet diarrhea through MAPK,IL 6 and Toll-like receptor signal pathway in the miRNAs-MUC13 regulation system.This study explored 4 molecular complex groups and 2 core genes(SRF and STAG 2)around the miRNAs-MUC 13 regulatory system,and carried out experimental verification,which provided theoretical reference and scientific basis for targeted treatment of piglet diarrhea.

关 键 词：Mucin 13 MICRORNA 仔猪腹泻 基因网络 

分 类 号：S828[农业科学—畜牧学]