摘要
目的分析小鼠脂肪组织中的长链非编码RNA(lncRNA)差异表达谱,探讨lncRNA Gm15290在脂肪生成中的调控作用。方法采用lncRNA微阵列分析WT和ob/ob小鼠脂肪组织中lncRNA的差异表达,通过Gene Ontology和DAVID数据库进行GO生物功能和基于KEGG的pathway富集分析。分离培养原代前脂肪细胞并用鸡尾酒法诱导分化脂肪细胞,采用pAd-Gm15290过表达或Gm 15290 siRNA转染脂肪细胞,Western blot检测成脂分子PPARγ、aP2及C/EBPα蛋白表达。结果lncRNA芯片检测显示,WT小鼠与ob/ob小鼠存在2246个差异表达lncRNA,其中53.6%(1204个)为外显子lncRNA,19.4%(436个)为内含子lncRNA,27.0%(606个)为反义lncRNA。采用GO和KEGG分析结果提示,lncRNA Gm15290与代谢和细胞生物合成过程有关。Gm15290过表达可以显著促进脂肪细胞分化,而Gm15290基因干扰明显抑制脂肪细胞分化,lncRNA Gm15290上调PPARγ、C/EBPα和aP2蛋白的表达水平。结论WT小鼠和ob/ob小鼠脂肪组织中的lncRNA表达谱不同,lncRNA Gm15290通过上调PPARγ、C/EBPα和aP2蛋白的表达来调节脂肪的生成。
Objective To analyze the differential expression profiles of long-chain non-coding RNA(lncRNA)in adipose tissue of mice and explore the regulation of lncRNA Gm15290 in adipogenesis.Methods The lncRNA microarray was used to analyze the differential expression of lncRNA in adipose tissue of WT and ob/ob mice.GO biological function and pathway enrichment based on KEGG were analyzed by gene ontology and David database.Primary preadipocytes were isolated and cultured,and induced to differentiate into adipocytes by cocktail method.After overexpression of pAd-Gm15290 or transfection of Gm15290 siRNA into adipocytes,the expression of PPAR gamma,aP2 and C/EBPαwas detected by Western blot.Results A total of 2246 lncRNAs were differentially expressed between WT mice and ob/ob mice by lncRNA chip.Of these differentially expressed lncRNAs,53.6%(1204)were exon lncRNA,19.4%(436)were intron lncRNA,and 27.0%(606)were antisense lncRNA.GO and KEGG analysis showed that lncRNA Gm15290 was involved in metabolic and cellular biosynthesis processes in these differentially expressed lncRNAs.Gm15290 overexpression significantly promoted adipocyte differentiation,while Gm15290 knockout significantly inhibited adipocyte differentiation.The lncRNA Gm15290 upregulated PPARγ,C/EBPαand aP2 protein expression.Conclusion The expression profiles of lncRNA in adipose tissue of WT mice and ob/ob mice are different.The lncRNA GM 15290 regulates adipogenesis by upregulating the expression of PPARγ,C/EBPαand aP2 protein.
作者
刘伟华
尹春燕
常明
LIU Weihua;YIN Chunyan;CHANG Ming(Department of Pediatrics,Xi’an First People’s Hospital,Xi’an 710004,China;Department of Pediatrics,Second Affiliated Hospital of Xi’an Jiaotong University;Department of Pediatrics,Ankang People’s Hospital)
出处
《山西医科大学学报》
CAS
2020年第2期147-152,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81803262)
陕西省自然科学基金资助项目(2016JM8134,2018JM7136)
西安市科技局卫生计划项目(J20170201)。
