旨在分析饲喂青贮黄梁木替代50%青贮玉米的川中黑山羊肝转录组测序中差异表达的mRNA、lncRNA及其靶基因,挖掘饲喂青贮黄梁木影响川中黑山羊肝代谢的关键基因,从而探究青贮黄粱木的饲喂效果.试验共选取6只雄性健康川中黑山羊[5月龄,平均体重(21.07±4.05)kg]并随机分为两组,试验组为青贮黄梁木替代50%青贮玉米组,对照组为青贮玉米组.试验期间采用全混合日粮(TMR)进行饲喂.于试验期的最后1d对山羊进行屠宰并取其肝,提取肝组织的RNA,构建文库后使用高通量测序技术进行转录组测序,以P<0.05为阈值筛选出差异mRNA和lncRNA,进行生物学功能富集分析并构建靶向关系网络.再从差异表达lncRNA和mRNA中各随机选择3个基因,采用RT-qPCR进行验证分析.结果表明,共发现差异表达的mRNA有1 696个,其中943个上调,753个下调;差异表达lncRNA共有33个,其中22个上调,11个下调.结合差异表达lncRNA的靶基因与差异表达mRNA的交集基因的富集分析发现,NDUFA10、NDUFB11、NDUFS5参与肝功能相关的信号通路,如多糖代谢过程、葡聚糖分解代谢过程、纤维素分解代谢等.结合lncRNA靶基因预测发现,GSTA3、GSTA4基因参与肝功能相关的信号通路如药物代谢-细胞色素P450及细胞色素P450对异源物质的代谢等代谢通路.本研究发现,在饲喂青贮黄梁木川中黑山羊的肝组织中,NDUFA10、NDUFB 11、NDUFS5以及GSTA3、GSTA4等可能从营养物质代谢和毒素代谢等方面影响肝功能.
The purpose of this study was to analyze the differentially expressed mRNA,lncRNA and its target genes in liver transcripome sequencing of Chuanzhong Black Goats fed silage Neolamarckia Cadamba instead of 50%silage corn,and to find out the key genes affecting liver metabolism of Chuanzhong Black Goats fed silage Neolamarckia Cadamba,so as to explore the feeding effect of silage Neolamarckia Cadamba.A total of 6 healthy male Chuanzhong Black Goats[5 months old,average body weight(21.07+4.05)kg]were selected and randomly divided into two groups:experimental group was fed silage Neolamarckia Cadamba replaced 50%silage corn,and control group was fed silage corn.Total mixed diet(TMR)was used for feeding during the experiment.On the last day of the experiment period,goats were slaughtered and their livers were taken.RNA from the liver tissues was extracted to construct a library.High-throughput sequencing technology was used for transcriptome sequencing.Differential mRNA and lncRNA were screened at a threshold of P<0.05,and biological function enrichment analysis was conducted to construct a targeted relationship network.Finally,3 genes were randomly selected from differentially expressed lncRNA and mRNA separately,and verified by RT-qPCR.The results showed that a total of 1 696 mRNA were differentially expressed,of which 943 were up-regulated and 753 were down-regulated.A total of 33 lncRNA were differentially expressed,of which 22 were up-regulated and 11 down-regulated.Combined with the enrichment analysis of target genes of differentially expressed lncRNA and intersection genes of differentially expressed mRNA,it was found that NDUFA10,NDUFB11 and NDUFS5 were involved in signal pathways related to liver function,such as polysaccharide metabolism,glucan catabolism and cellulose catabolism.Combined with lncRNA target gene prediction,it was found that GSTA3 and GSTA4 genes were involved in signal pathways related to liver function,such as drug metabolism-cytochrome P450 and cytochrome P450 metabolism of heterologous substances.In conclusion,NDUFA10,NDUFB11,NDUFS5,GSTA3 and GSTA4 may affect the liver function from the aspects of nutrient metabolism and toxin metabolism in the liver tissue of Chuanzhong Black Goats fed with silage Neolamarckia Cadamba.