黄芪多糖对C3H10T1/2细胞棕色脂肪分化中长链非编码RNA表达谱的影响  

The Effects of Astragalus Polysacharin on the Long Non-coding RNA Expression Profiles during Brown Adipogenesis of C3H10T1/2 Cell

作　　者：张世和 曹宇鑫 邓步皓 高旭阳 赵俊星[1] ZHANG Shihe;CAO Yuxin;DENG Buhao;GAO Xuyang;ZHAO Junxing(College of Animal Science and Veterinary Medicine,Shanxi Agricultural University,Taigu 030800,China)

机构地区：[1]山西农业大学动物科技学院,太谷030800

出　　处：《激光生物学报》2020年第4期333-343,351,共12页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(31972559);山西省2018年“1331”工程项目(1811331)。

摘　　要：本试验旨在探究黄芪多糖(APS)对小鼠间充质干细胞C3H10T1/2细胞棕色脂肪化过程中长链非编码RNA(lncRNA)表达谱的影响。试验以C3H10T1/2细胞为研究对象,在成脂诱导分化培养液中添加0.4 g/L的APS,以诱导分化1.5 d的细胞构建文库后测序,筛选差异mRNAs和差异lncRNAs,并对差异mRNAs和差异lncRNAs的顺式及共表达作用预测的靶基因进行了功能分析。通过荧光定量PCR随机分析了3个lncRNAs和3个mRNAs的表达水平,验证了测序结果的准确性。研究结果表明,本次测序共得到13450个lncRNAs和57776个mRNAs。通过对其表达量、长度、外显子个数和成对重复性检验分析证明了测序数据的可靠性较好。筛选共得到153个差异表达的lncRNAs和1238个差异表达的mRNAs。结果表明,差异mRNAs主要基因本体(GO)注释富集在53个功能分类中,京都基因与基因组百科全书(KEGG)分析富集在343条通路中。差异lncRNAs顺式及共表达作用预测的靶基因GO注释分别富集在34和33个功能分类中,在分子功能中条目一致。KEGG分析显示,多个基因富集在脂肪代谢和脂肪分化的信号通路中,尤其在胰高血糖素及cAMP信号通路中富集显著。综上表明,APS导致了C3H10T1/2细胞成脂分化中lncRNA表达谱的变化。本研究结果可为进一步解析APS对干细胞的分化调节提供科学依据。The objective of the present study was to investigate the effects of astragalus polysacharin(APS)on the expression profile of long non-coding RNA(lncRNA)during brown adipogenesis of mouse C3H10T1/2 cells.0.4 g/L APS was added to induction medium,and the 1.5 day brown adipogenic induced cells were used for the library construction and sequence,we screened differential mRNAs and differential lncRNAs,and analyzed the differential mRNAs and the target genes predicted by differential lncRNAs cis and co-expression were analyzed functionally.In addition,the expression levels of three random lncRNAs and three mRNAs were detected by fluorescence quantitative PCR to verify the accuracy of the sequencing results.The results indicated that a total of 13450 lncRNAs and 57776 mRNAs were obtained by sequencing.The reliability of the sequencing data was good by analyzing the expression amount,length,number of exons and pair repeatability.A total of 153 differentially expressed lncRNAs and 1227 differentially expressed mRNAs were screened The results showed that the differential mRNAs GO annotation was enriched in 53 functional categories,and the KEGG analysis was enriched in 343 pathways.The GO annotations of target genes predicted by cis and co-expression of differential lncRNAs were enriched in 34 and 33 functional categories,respectively,and the molecular function items were the same;KEGG analysis showed that many genes were enriched in the signal pathway of fat metabolism and fat differentiation,especially in glucagon and cAMP signal pathway.In conclusion,APS could effectively affect lncRNA expression profile of brown adipogenic induced C3H10T1/2 cell,which will further provide fundamental data for understanding the effect of APS on stem cell differentiation.

关 键 词：黄芪多糖 lncRNAs C3H10T1/2 棕色脂肪分化 表达谱 

分 类 号：S813.3[农业科学—畜牧学]