沙葱提取物对杜寒杂交羊肌肉脂代谢相关基因表达及甲基化的影响  

Effects of Allium mongolicum Regel Extracts on Lipid Metabolism Related Gene Expression and Methylation in Muscle of Dorper×Thin-Tailed Han Crossbred Sheep

作　　者：范泽军 敖长金[1] 杜红喜 包志碧 赵亚星 FAN Zejun;AO Changjin;DU Hongxi;BAO Zhibi;ZHAO Yaxing(College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学动物科学学院,呼和浩特010018

出　　处：《动物营养学报》2019年第5期2340-2349,共10页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：国家自然科学基金项目(31460611)

摘　　要：本试验旨在研究沙葱提取物对杜寒杂交羊肌肉脂代谢相关基因表达及甲基化的影响。采用单因素完全随机设计,选取60只体重(35～40 kg)相近、4.5月龄的杜寒杂交母羊,随机分为4组,每组15只。对照组(T1组)饲喂基础饲粮,试验组在基础饲粮中每只分别添加沙葱粉(10.0 g/d,T2组)、沙葱水溶性提取物(3.2 g/d,T3组)、沙葱脂溶性提取物(2.8 g/d,T4组)。预试期为15 d,正试期为60 d。正试期结束后,每组随机选取3只羊屠宰,采集背最长肌样品。采用目的区域甲基化水平测序(二代测序)检测乙酰辅酶A羧化酶α(ACACA)、硬脂酰辅酶A去饱和酶(SCD)、固醇调节元件结合转录因子1(SREBF1)基因转录起始位点上游2K至第一外显子下游1K区域内胞嘧啶-磷酸-鸟嘌呤(CpG)岛甲基化水平,实时荧光定量PCR法测定其基因相对表达量。结果表明:1)试验第1～30天,T2组干物质采食量显著低于其他3组(P <0.05)。2)与T1组相比,T2组、T3组、T4组ACACA、SCD基因相对表达量均有不同程度的提高,其中T3组>T2组>T4组>T1组。T3组ACACA、SCD、SREBF1基因相对表达量均显著高于T1组(P <0. 05),T2组ACACA基因相对表达量显著高于T1组(P <0. 05),T3组ACACA、SREBF1基因相对表达量显著高于T4组(P<0.05)。3) ACACA、SCD、SREBF1基因各CpG岛及位点均呈现高度去甲基化状态,且各组间CpG岛的平均甲基化水平差异不显著(P> 0.05)。ACACA、SCD、SREBF1的基因表达与CpG岛甲基化水平不相关(P>0.05)。综上所述,沙葱及其提取物可以上调杜寒杂交羊背最长肌ACACA、SCD、SREBF1的基因表达,饲粮添加沙葱水溶性提取物的上调幅度最大,沙葱粉次之,沙葱脂溶性提取物最小,且这种上调与特定区域的DNA甲基化修饰无关。This experiment was conducted to investigate the effects of Allium mongolicum Regel extracts on lipid metabolism related gene expression and methylation in muscle of Dorper×thin-tailed Han crossbred sheep.A single factor complete random design was used in the current study,sixty 4.5-month-old Dorper×thin-tailed Han crossbred ewes with similar body weight(35 to 40 kg)were assigned to 4 groups with 15 ewes in each group.Sheep in the control group(T1 group)were fed a basal diet,and the others in experimental groups were fed the basal diet supplemented with Allium mongolicum Regel powder(10.0 g/d each sheep,T2 group),Allium mongolicum Regel water-soluble extract(3.2 g/d each sheep,T3 group),Allium mongolicum Regel fat-soluble extract(2.8 g/d each sheep,T4 group),respectively.The pre-trial lasted for 15 days,and the trial lasted for 60 days.After the trial,three sheep were randomly selected from each group and slaughtered to collect the longissimus dorsi muscle samples.The cytosine-phosphoric acid-guanine(CpG)island methylation level of acetyl-CoA carboxylaseα(ACACA),stearoyl-CoA desaturase(SCD)and sterol regulatory element binding transcription factor 1(SREBF1)genes from 2K upstream of transcriptional start site to 1K downstream of the first exon were measured according to methylation level sequencing of target region(second generation sequencing),and the gene relative expression levels were determined by real-time quantitative polymerase chain reaction.The results showed as follows:1)during the 1 to 30 days of the experiment,the dry matter intake in T2 group was significantly lower than that in other 3 groups(P<0.05).2)Compared with T1 group,the relative gene expression levels of ACACA and SCD in T2 group,T3 group and T4 group were increased at different degrees,among them,T3 group>T2 group>T4 group>T1 group.The relative expression levels of ACACA,SCD and SREBF1 genes in T3 group were significantly higher than those in T1 group(P<0.05),the relative expression level of ACACA gene in T2 group was significantly high

关 键 词：沙葱提取物 杜寒杂交羊 脂肪代谢 甲基化 基因表达 表观遗传 

分 类 号：S826[农业科学—畜牧学]