基于高通量转录组测序的辣木叶粉对小鼠肝脏差异表达基因的影响

Effect of Moringa oleifera on Mice Liver Transcriptome Based on High-throughput RNA-seq Technology

【目的】辣木(Moringa oleifera Lam.)是一种营养价值丰富的新型食品或饲料资源,但是食用或饲用辣木后对动物机体代谢的调控机制仍不清楚。【方法】18只小鼠随机分为2组,即辣木叶粉饲料组(50%MO)和基础饲料组(对照组)。在饲料干预的第3、10和25天每组随机取3只小鼠进行解剖,采集肝脏组织样品,并利用高通量RNA-seq测序技术对小鼠肝脏进行RNA测序,使用Top Hat v2.0.13软件将测序得到的reads序列与小鼠(GRCm38)参考基因组序列比对,计算表达量并鉴定差异表达基因,同时对差异表达基因参与的GO功能及代谢途径进行富集。【结果】从18个样本中共获得转录组数据约109.063 Gb,1 136 175 196条clean reads;与对照组相比,辣木叶粉饲料组表达上调和下调的差异表达基因共有1 227个;GO功能分类注释发现,第3、10和25天的差异基因都主要集中在细胞过程(cellular process)、细胞(cell)和结合(binding)3个过程;差异基因显著富集的KEGG代谢通路有37条(P<0.05)。【结论】辣木叶粉对小鼠肝脏中的过氧化物酶体、病毒性心肌炎和类固醇生物合成等代谢通路有一定影响。

[Purpose]Moringa oleifera Lam.is a nutritious new food or feed resource.However,the effect of M.oleifera on the metabolism mechanism of the body is still unclear.[Method]We detected the differentially expressed genes and metabolism pathways between M.oleifera diet group and control group in 18 mice.The tested mice were slaughtered after the test period of 3,10 and 25 days,and liver tissues were isolated and their mRNA was extracted,respectively.The liver transcriptomes were analyzed by high-throughput RNA sequencing system.All RNA-Seq reads were mapped on the reference mouse genome(GRCm38)using TopHat v2.0.13 software.We calculated the expression level and identified the differentially expressed genes.The genes were annotated using GO and KEGG databases.[Results]A total of 1 136 175 196 reads were obtained from the liver of mice.Compared with the control group,there were 1 227 differentially expressed genes that up-regulated or down-regulated in 50%MO group.Using gene ontology analysis,three gene ontologies were found to be enriched in the genes with different expression level:cellular process,cell and binding.The differentially expressed genes were significantly enriched 37 pathway in the KEGG database(P<0.05).[Conclusion]There were a certain effect of M.oleifera leaves powder in the peroxisomes,viral myocarditis,steroid biosynthesis and other metabolic pathways in mice.