多烯磷脂酰胆碱干预大鼠非酒精性脂肪肝细胞模型基因芯片组学分析  

Gene chipset analysis in rats with non-alcoholic fatty liver disease intervened by polyene phosphatidylcholine

作　　者：刘锐[1] 何嘉 刘秀芳 冯晓秋[1] 古展鑫 康善平[1] Liu Rui;He Jia;Liu Xiufang;Feng Xiaoqiu;Gu Zhanxin;Kang Shanping(Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,Guangxi Zhuang Autonomous Region,China;Graduate School of Guangxi University of Chinese Medicine,Nanning 530001,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西中医药大学附属瑞康医院,广西壮族自治区南宁市530011 [2]广西中医药大学研究生学院,广西壮族自治区南宁市530001

出　　处：《中国组织工程研究》2018年第36期5833-5839,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区项目(81460717);广西自然科学基金面上项目(2014GXNSFAA118249)~~

摘　　要：背景:随着信息技术的发展及与生物医学领域的互融,产生了大量基因组学、转录组学等相关的大数据,并通过生物信息学分析与计算机科学相结合的方法对这些数据进行分析,可研究多个基因之间的相互联系。多烯磷脂酰胆碱保护肝脏、改善脂肪代谢、干预非酒精性脂肪肝细胞的相关靶点及其作用机制尚未完全明确。目的:通过筛选mRNA(ClariomTM S Assay)芯片及miRNA芯片的显著差异性基因,预测microRNA相关靶基因,研究多烯磷脂酰胆碱改善肝细胞脂肪代谢的调控机制。方法:(1)提取多烯磷脂酰胆碱灌胃SD大鼠的含药血清;(2)培养BRL大鼠肝细胞,加含体积分数50%胎牛血清的培养基建立非酒精性脂肪肝细胞模型,24h后模型组加入正常血清、多烯磷脂酰胆碱组加入多烯磷脂酰胆碱含药血清进行干预;(3)提取细胞总RNAs,采用ClariomS表达谱芯片及miRNA芯片检测细胞mRNA及miRNA的表达水平。运用生物信息学方法预测miRNA的靶基因,对脂肪代谢相关基因进行富集分析和通路分析。结果与结论:经筛选和预测得到5 629个预测靶基因参与13种生物学过程,14种细胞成分,有11种分子作用,代谢途径、嘌呤代谢、类固醇生物合成路径被富集;rno-miR-21-5p、rno-miR-370-3p、rno-miR-542-3p为调控网络中心;IPA数据库共检出56个与实验有关的脂肪代谢基因。提示:多烯磷脂酰胆碱治疗非酒精性脂肪肝的机制可能与rno-miR-21-5p、rno-miR-370-3p、rno-miR-542-3p,代谢途径、嘌呤代谢、类固醇生物合成路径相关性较高。BACKGROUND:With the development of information technology and integration with the biomedical field,a large number of big data related to genomics and transcriptomics,have been generated,and these data are analyzed through a combination of bioinformatics and computer science.Further the interrelationships between multiple genes are explored.The targets of polyene phosphatidylcholine protecting liver,improving lipid metabolism and intervening nonalcoholic fatty liver and underlying mechanisms remain unclear.OBJECTIVE:To predict and investigate the regulatory mechanism of polyene phosphatidylcholine improving fat metabolism of hepatocyte based on screening the difference between mRNA(Clariom?S Assay)and miRNA expression.METHODS:Serum of Sprague-Dawley rats receiving intragastric administration of polyene phosphatidylcholine was extracted.BRL rat hepatocytes were cultured in the medium containing 50%of fetal bovine serum to establish the model of nonalcoholic fatty hepatocytes.Then polyene phosphatidylcholine serum was used as experimental group and negative control serum was used as control group,respectively.Total RNAs were extracted and differently expressed miRNAs were detected by Clariom S expression profile chip for expression level of mRNA and miRNA.Bioinformatics method was used to predict target genes of differential miRNAs regulation.Enrichment of functions and signaling pathways of the target genes was conducted by fat metabolism related genes.RESULTS AND CONCLUSION:In screening and predicting,5 629 target genes were involved in 13 kinds of biological processes,14 kinds of cellular components,and 11 kinds of molecular functions.Metabolic pathways,purine metabolism and steroid biosynthesis pathway were enriched.rno-miR-21-5p,rno-miR-370-3p,rno-miR-542-3p were the center of the molecular network.According to IPA database,56 genes with fat metabolism effective interaction were screened out.These results show rno-miR-21-5p,rno-miR-370-3p,rno-miR-542-3p and Metabolic pathways,purine metabolism and steroid bios

关 键 词：多烯磷脂酰胆碱 非酒精性脂肪肝 miRNA 靶基因 脂肪代谢 生物信息学分析 组织构建 

分 类 号：R318[医药卫生—生物医学工程]