食品科学 ›› 2022, Vol. 43 ›› Issue (13): 140-145.doi: 10.7506/spkx1002-6630-20210319-237

• 营养卫生 • 上一篇    

内源性n-3多不饱和脂肪酸对下丘脑神经干细胞来源外泌体miRNA的调节

路宗博,张盈月,葛科立,张金玉,薛美兰,葛银林   

  1. (青岛大学基础医学院,山东 青岛 266073)
  • 发布日期:2022-07-27
  • 基金资助:
    山东省重点研发项目(2017GSF218084);青岛市应用基础研究项目(19-6-2-31-cg); 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)

Regulation of Endogenous n-3 Polyunsaturated Fatty Acids on Exosomal miRNAs Derived from Hypothalamic Neural Stem Cells

LU Zongbo, ZHANG Yingyue, GE Keli, ZHANG Jinyu, XUE Meilan, GE Yinlin   

  1. (School of Basic Medicine, Qingdao University, Qingdao 266073, China)
  • Published:2022-07-27

摘要: 目的:通过培养fat-1转基因小鼠下丘脑神经干细胞获取外泌体并提取总RNA进行高通量测序,研究内源性n-3多不饱和脂肪酸(n-3 polyunsaturated fatty acids,n-3 PUFAs)对丘脑神经干细胞来源外泌体miRNA的调节作用,探究n-3 PUFAs抑制下丘脑炎症及肥胖的作用机制。方法:分别培养fat-1转基因小鼠与野生型C57BL/6小鼠下丘脑神经干细胞,从细胞培养上清液中收集外泌体并提取总RNA,经高通量测序后利用生物信息学技术对差异表达的miRNA进行分析及靶基因预测,对靶基因进行基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析后,利用实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qPCR)进行相对表达量的测定。结果:与野生型C57BL/6小鼠相比,fat-1转基因小鼠下丘脑神经干细胞来源的外泌体miRNA具有特异表达谱,且具有显著差异表达的miRNA靶基因处于下丘脑炎症、脂肪酸代谢等通路中关键位置。经qPCR验证,差异表达的miRNA关键靶基因表达均受到调控。结论:内源性n-3 PUFAs水平的增加可以调节下丘脑神经干细胞外泌体中miRNA的表达,进而调控炎症反应与脂质代谢相关基因的表达,从而发挥抑制下丘脑炎症、抵抗肥胖的作用。

关键词: 内源性n-3多不饱和脂肪酸;外泌体;miRNA;下丘脑炎症;脂质代谢

Abstract: Exosomes were obtained by cultivating hypothalamic neural stem cells from fat-1 transgenic mice and total RNA was extracted from them for high-throughput sequencing. To study the regulatory effect of n-3 polyunsaturated fatty acids (n-3 PUFAs) on exosomal miRNAs derived from neural stem cells in the hypothalamus, and to explore the mechanism of action of n-3 PUFAs in inhibiting hypothalamic inflammation and obesity, the hypothalamic neural stem cells from fat-1 transgenic mice and wild-type C57BL/6 mice were cultured, separately. Exosomes were collected from the cell culture supernatant for total RNA extraction. After high-throughput sequencing, bioinformatics was used to analyze the differentially expressed miRNAs and predict the target genes. The target genes were analyzed by KEGG (Kyoto encyclopedia of genes and genomes) enrichment analysis and their relative expression levels were detected by quantitative real-time polymerase chain reaction (qPCR). The results showed that compared with wild-type C57BL/6 mice, the exosome miRNAs derived from hypothalamic neural stem cells in fat-1 transgenic mice had distinctive expression profiles, and the target genes of the miRNAs that were significantly differentially expressed were at key positions in the hypothalamic inflammation and fatty acid metabolism pathways. As validated by qPCR, the expression of the key target genes that were differentially expressed were regulated by n-3 PUFAs. In conclusion, the increase of endogenous n-3 PUFAs level can regulate the expression of miRNAs in exosomes from neural stem cells in the hypothalamus and the expression of genes related to inflammatory response and lipid metabolism, thereby inhibiting hypothalamic inflammation and resisting obesity.

Key words: endogenous n-3 polyunsaturated fatty acids; exosome; miRNA; hypothalamic inflammation; lipid?metabolism

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