高剂量酒糟摄入对山羊肾脏组织氧化指标、炎性因子、转录谱的影响

Effects of high dose distillers grains on oxidative index,inflammatory factor,transcript profile in kidney of goats

旨在探索高剂量酒糟摄入对山羊肾脏组织氧化指标、炎性因子、转录谱的影响,为揭示高剂量酒糟饲喂对山羊肾脏损伤的机制提供理论基础。选取6只30日龄荣昌本地山羊随机分为2组,每组3只,对照组按照《肉羊饲养标准》(NY/T816-2004)推荐的山羊饲养标准饲喂;试验组用酒糟代替日粮中75%蛋白质和能量成分,试验期为90 d。90 d后采集山羊肾脏组织,利用生化法测定山羊肾脏组织T-SOD、CAT、GSH-Px、XOD、T-AOC含量,用ELISA法测定山羊肾脏组织IL-6、IL-1β、TNF-α含量,用RNA-Seq技术进行转录组测序分析。结果显示,试验组XOD水平显著低于对照组(P<0.05),T-SOD和T-AOC水平极显著低于对照组(P<0.01)。试验组IL-1β水平极显著高于对照组(P<0.01),IL-6、TNF-α水平显著高于对照组(P<0.05)。与对照组比较,试验组共发现546个差异表达基因,其中有385个基因表达上调,161个基因表达下调,qRT-PCR显示挑选的基因与转录组测序结果的表达趋势具有一致性。GO富集分析显示试验组差异表达基因在生物过程中富集的主要类别是电子传递链、呼吸电子传递链;在分子功能中,主要类别是转运活性、跨膜转运蛋白活性;在细胞成分中,主要类别是膜的整体组成、膜的固有成分。KEGG分析显示试验组差异表达基因主要富集在生热作用、帕金森病、氧化磷酸化等通路中。转录组测序与ELISA、生化试验结果具有一致性,均表明75%酒糟饲喂山羊造成肾脏发生氧化应激和炎性损伤。

This study aims to explore the effects of high dose distillers grains on oxidative index,inflammatory factor,transcript profile in kidney of goats,and to provide a theoretical basis for revealing the mechanism of kidney injury induced by high dose distillers grains feeding in goats.Six 30-day-old Rongchang local goats were randomly divided into 2 groups with 3 goats in each group.The c.ontrol group was fed according to the goat feeding standard recommended by ifeeding standard of meat-producing sheep and goats'(NY/T816-2004),and the experimental group was fed with distillers grains instead of 75% protein and energy components in the diet.The experiment lasted for90 days.After 90 days,goat kidney tissues were collected.The contents of T-SOD,CAT,GSH-Px,XOD and T-AOC were determined by biochemical method.The contents of IL-6,IL-1β and TNF-α were determined by ELISA.Transcriptome sequencing analysis was performed by RNA-Seq technology.The results showed that the level of XOD in experimental group was significantly lower than that in control group(P<0.05),and the levels of T-SOD and T-AOC were extremely significantly lower than those in control group(P<0.01).The levels of IL-1β in experimental group were extremely significantly higher than those in control group(P<0.01),and the levels of IL-6and TNF-α in experimental group were significantly higher than those in control group(P<0.05).Compared with the control group,a total of 546 differentially expressed genes were found in the experimental group,among them,385 genes were up-regulated and 161 genes were down-regulated.GO enrichment analysis showed that the main types of differentially expressed genes in the experimental group enriched in biological processes were electron transport chain and respiratory electron transport chain.In molecular function category,the main proteins have transport activity and transmembrane transport protein activity.Among the cell components,the main categories were the integral component of membrane and intrinsic component of membrane.KEGG analysis showed that the differentially expressed genes in the experimental group were mainly enriched in thermogenesis,Parkinson's disease,oxidative phosphorylation and other pathways.The results of transcriptome sequencing were consistent with those of ELISA and biochemical experiments,which showed that feeding with 75% distillers grains caused oxidative stress and inflammatory damage to their kidneys.