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分别提取灌喂氧化鱼油以及鱼油的黄颡鱼胃肠道肠道黏膜的总RNA。将氧化鱼油组、鱼油组总RNA等量混合后,采用RNA-Seq测序,用Trinity进行de novo拼接、组装,对单一基因进行功能注释;再将氧化鱼油组、鱼油组总RNA分别测序、注释后进行基因表达量分析,并计算氧化鱼油组对鱼油组单一基因的差异表达定量分析,以log_2(OFH/FH)值代表氧化鱼油组相对于鱼油组基因的差异表达量。基因差异表达显示,氧化鱼油导致黄颡鱼胃肠道黏膜组织中胆固醇、胆汁酸生物合成代谢途径的酶以及涉及胆固醇和胆汁酸吸收转运蛋白基因差异表达。黄颡鱼胃肠道黏膜中以乙酰辅酶A为原料的胆固醇生物合成途径关键酶基因差异表达显著下调,从细胞外吸收转运胆固醇的主要蛋白基因差异表达下调,显示灌喂氧化鱼油导致黄颡鱼胃肠道黏膜胆固醇合成能力下降、从其他组织吸收胆固醇的能力下降,血清胆固醇含量下降4.06%。灌喂氧化鱼油后,黄颡鱼胃肠道黏膜以胆固醇为原料的初级胆汁酸合成代谢的关键酶差异表达显著上调,而胆汁酸转运到细胞外和转运到肠腔的载体蛋白基因差异表达下调,肠道再吸收胆汁酸的转运载体蛋白基因差异表达下调及肝细胞从血液吸收转运的载体蛋白基因差异表达上调,而血清胆汁酸含量下降了20.00%。结果表明,胆汁酸的肠肝循环发生障碍,肠腔和血清胆汁酸含量下降、胆汁酸在细胞出现淤积的趋势。
Total RNA extracted from gastrointestinal mucosa of Catfish fed with oxidized fish oil and fish oil respectively. The total amount of total RNA of the oxidized fish oil group and the fish oil group was mixed by RNA-Seq sequencing, de novo splicing and assembly with Trinity, and functional annotation of the single gene. Sequencing and annotating the total RNA of the oxidized fish oil group and the fish oil group respectively The gene expression was analyzed and the differential expression of single gene in fish oil group was calculated. The log 2 (OFH / FH) value was used to represent the differential gene expression of oxidized fish oil group relative to fish oil group. Gene differential expression shows that oxidized fish oil causes the cholesterol, bile acid biosynthetic metabolic pathway enzymes in the gastrointestinal mucosa of the catfish and the differential expression of genes involved in cholesterol and bile acid uptake transporters. The differences of the expression of key genes of cholesterol biosynthesis pathway from acetylcholinesterase in the intestinal mucosa of C. anguillarum were significantly down-regulated. The differential expression of the major protein genes that transported and transported cholesterol from the extracellular matrix was down-regulated. Gastrointestinal mucosal cholesterol synthesis decreased ability to absorb cholesterol from other tissues decreased serum cholesterol decreased by 4.06%. After fed with oxidized fish oil, the differential expression of key enzymes of primary bile acid anabolism in the gastrointestinal mucosa of Chub mackerel with cholesterol as raw material was significantly upregulated, whereas the expression of the carrier protein gene of bile acids translocated to the extracellular and transported to the intestine was down-regulated , The intestinal reabsorption of bile acid transporter protein gene expression downregulation and liver cells from the blood absorption and transport of the carrier protein gene differential expression was increased, and serum bile acid content decreased by 20.00%. The results showed that the bile acids in the enterohepatic circulation obstacles, intestine and serum bile acid content decreased, bile acid deposition in the cell trend.