睾酮缺乏对高脂诱导小型猪内脏脂肪组织DNA甲基化的影响  

Effects of Testosterone Deficiency on DNA Methylation in Visceral Adipose Tissue ofMiniature Pigs Fed on a High Fat Diet

作　　者：李庆海[1] 邓利群 凌云[2] 蔡月琴[2] 黄俊杰 章学东[1] 蔡兆伟[2] Li Qinghai;Deng Liqun;Ling Yun;Cai Yueqin;Huang Junjie;Zhang Xuedong;Cai Zhaowei(Hangzhou Academy of Agricultural Sciences,Hangzhou,310024;Laboratory Animal Research Center/Institute of Comparative Medicine,Zhejiang Chinese Medical University,Hangzhou,310053)

机构地区：[1]杭州市农业科学研究院,杭州310024 [2]浙江中医药大学动物实验研究中心,比较医学研究所,杭州310053

出　　处：《基因组学与应用生物学》2021年第3期1077-1084,共8页Genomics and Applied Biology

基　　金：浙江省自然科学基金项目(LY18C040003);浙江省公益技术应用研究项目(2018C37084)共同资助。

摘　　要：为探讨DNA甲基化在睾酮缺乏促进高脂饮食诱导的小型猪肥胖中的作用,本研究采用甲基化DNA免疫共沉淀测序(MeDIP-Seq)技术分析高脂饲喂的不去势、去势和去势+睾酮3组小型猪内脏脂肪组织DNA甲基化差异,对筛选出的差异甲基化基因进行注释和功能富集分析,并运用RT-qPCR技术检测差异甲基化基因的表达。结果表明,不去势、去势和去势+睾酮3组样本在基因组上的甲基化分布情况相似,即Gene body区的甲基化水平高于3'UTR和5'UTR区。另外,在去势vs.不去势和去势vs.去势+睾酮两组样本中分别筛选得到2839个和2510个差异甲基化基因,这些基因主要富集在脂肪细胞因子转导、抗原处理以及呈递、脂肪酸代谢和氨基酸代谢等通路。睾酮缺乏导致高脂饲喂小型猪内脏脂肪组织LEP以及NCF1内含子区甲基化和SLC-27A1启动子区甲基化水平升高,并且LEP和NCF1 mRNA表达与甲基化呈正相关,而SLC27A1 mRNA表达与甲基化呈负相关。本研究推测,睾酮缺乏可能通过影响脂质代谢和炎症反应等多个途径基因DNA甲基化参与调控脂肪沉积和肥胖发生。To explore the role of DNA methylation in the effects of testosterone deficiency induced obesity in miniature pigs fed on a high-fat diet(HFD),the study employed methylated DNA immunoprecipitation sequencing(Me-DIP-Seq)technology to examine DNA methylation alterations in the visceral adipose tissue(VAT)of HFD-fed intact male pigs(SHAM),castrated male pigs(CAS)and castrated male pigs with testosterone treatment(CAS+T).Differentially methylated genes(DMGs)were screened out and used for functional enrichment analysis.RT-qPCR was used to detect the expression of DMGs.The results showed that the genome distribution of DNA methylation was similar among SHAM,CAS and CAS+T,with a relatively higher methylation level in the gene body region than that in the 3'UTR and 5'UTR.Furthermore,2839 and 2510 DMGs were obtained from CAS vs.SHAM and CAS vs.CAS+T,respectively.These DMGs were mainly enriched in adipocytokine signaling pathway,antigen processing and presentation,fatty acid metabolism and amino acid metabolism.Testosterone deficiency caused the increase of DNA methylation in LEP and NCF1 intron and in SLC27A1 promoter.Levels of LEP and NCF1 mRNA expression were positively correlated with their methylation,while the SLC27A1 mRNA level was negatively correlated with its methylation.Our study speculated that testosterone deficiency might regulate fat deposition and obesity through affectingDNAmethylation ofgenes involved in lipid metabolismand inflammatoryresponse related multiple pathways.

关 键 词：睾酮 小型猪 脂肪沉积 DNA甲基化 炎症 

分 类 号：R589.2[医药卫生—内分泌] R-332[医药卫生—内科学]