摘要
目的:研究体内n-3多不饱和脂肪酸(n-3 polyunsaturated fatty acids,n-3 PUFAs)含量的增加对小鼠体质量和血液中外泌体miRNAs表达的影响,探讨n-3 PUFAs通过外泌体抑制肥胖的作用机制。方法:利用能自发生成n-3 PUFAs的fat-1转基因小鼠和同窝野生型小鼠(对照),通过高脂饮食(high-fat diet,HFD)建立肥胖动物实验模型,测定小鼠体质量。提取小鼠血浆中的外泌体并鉴定;分离外泌体内的RNA,构建文库并进行miRNA高通量测序。根据测序结果通过生物信息学方法分析找到其调控的靶基因和相关联的通路,发现miRNA-靶基因互作关系。验证miRNA与肥胖的关联度以及在肥胖中所发挥的作用。结果:fat-1转基因小鼠体质量明显低于野生型;外泌体提取鉴定成功;miRNA高通量测序结果显示,不同小鼠组间进行对比时,差异表达显著(P<0.05且差异倍数(fold change,FC)≠1)的miRNA有46个;生物信息学分析发现6个重要miRNA(mmu-miR-665-3p、mmu-miR-122-5p、mmu-miR-122-3p、mmu-miR-194-5p、mmu-miR-34c-5p、mmu-miR-223-3p)落在脂肪酸代谢通路以及内吞通路关键位置,功能与脂质代谢和肥胖相关,所对应的靶基因分别为Fads1、Elovl2、Elov6、Hadha、Scad1、Scad2、Hsd17b12、Acot2、Acot4和Arf6、H2-T-ps、Arrb1、Ist1、H2-T10、Wwp1、Snx4、IL2rb、Mvb12b、Rab11、fip3、Kif5a、Nedd4l。结论:n-3 PUFAs含量的增加能够有效降低小鼠的体质量,抑制肥胖。n-3 PUFAs能够调节血液中外泌体内miRNA的表达,其中具有显著差异性的miRNA与肥胖有关,其相关的靶基因集中在脂质代谢相关的分子通路中,提示可能是n-3 PUFAs降低体质量抑制肥胖机制之一。
Objective:To investigate the effects of increasing n-3 polyunsaturated fatty acids(n-3 PUFAs)on the body mass and the expression of exosomal miRNAs in the blood of mice,and to explore the antiobesity mechanism of n-3 through the exosomes.Methods:fat-1 transgenic mice were fed a high-fat diet(HFD)to create an obese animal model and the wild-type littermates served as the control group.The body masses of mice were measured,and exosomes from the plasma of mice were extracted and identified.RNA was isolated from the exosomes for high-throughput sequencing of miRNAs and construction of a library.According to the sequencing results,the target genes and related signaling pathways were explored by bioinformatics analysis,and the relationship between miRNAs and their target genes was also elucidated.Finally,the association of miRNAs with obesity and its roles in obesity were validated.Results:The body mass of fat-1 transgenic mice was significantly lower than that of their wild-type counterparts.The exosomes were successfully extracted and identified.High-throughput sequencing showed 46 significantly differentially expressed miRNAs(P<0.05)with fold change(FC)value≠1.Bioinformatics analysis revealed that six important miRNAs(mmu-miR-665-3 p,mmu-miR-122-5 p,mmu-miR-122-3 p,mmu-miR-194-5 p,mmu-miR-34 c-5 p,and mmu-miR-223-3 p)were confirmed to play critical roles in fatty acid metabolism and endocytosis pathways,exerting functions related to lipid metabolism and obesity.Their target genes were Fads1,Elovl2,Elov6,Hadha,Scad1,Scad2,Hsd17 b12,Acot2 and Acot4;and Arf6,H2-T-ps,Arrb1,Ist1,H2-T10,Wwp1,Snx4,IL2 rb,Mvb12 b,Rab11,fip3,Kif5 a and Nedd4 l in the two pathways,respectively.Conclusion:Increasing n-3 PUFAs can reduce body mass and alleviate obesity in mice.In addition,n-3 PUFAs can regulate the expression of miRNAs in blood exosomes;the significantly differentially expressed miRNAs are related to obesity,and their target genes are mainly involved in fatty acid metabolism pathways,suggesting that n-3 PUFAs may be involved in the molecular mechanism for inhibiting obesity.
作者
吴聪
葛科立
路宗博
郑征
张金玉
薛美兰
葛银林
WU Cong;GE Keli;LU Zongbo;ZHENG Zheng;ZHANG Jinyu;XUE Meilan;GE Yinlin(School of Basic Medicine,Qingdao University,Qingdao 266021,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2021年第7期176-184,共9页
Food Science
基金
山东省自然科学基金面上项目(ZR2018MH009)
青岛市民生科技计划项目(19-6-1-47-nsh)。
作者简介
第一作者:吴聪(1994-)(ORCID:0000-0002-7651-5006),男,硕士研究生,研究方向为基因治疗。E-mail:wczqyc@163.com;通信作者:葛银林(1957-)(ORCID:0000-0003-4990-066X),男,教授,博士,研究方向为基因诊断与基因治疗。E-mail:geyinlin@126.com。
