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本实验研究了饲料氧化鱼油对草鱼肠道屏障结构和肠道通透性的影响。以豆油、鱼油、氧化鱼油作为饲料脂肪源,分别设计鱼油组(6F)、豆油组(6S)、2%氧化鱼油(2OF)、4%氧化鱼油(4OF)及6%氧化鱼油(6OF)5组等氮、等能半纯化饲料,在池塘网箱养殖草鱼[平均体质量(74.8±1.2)g]72 d。采用实时荧光定量PCR(qRT-PCR)的方法,测定了草鱼肠道紧密连接蛋白中闭锁蛋白Occludin,闭合蛋白Claudin-3、Claudin-15a和胞浆蛋白ZO-1、ZO-2、ZO-3的基因表达活性,并结合肠黏膜细胞间紧密连接结构透射电镜观察结果、肠道通透性标志性指标综合分析。结果显示:在添加氧化鱼油后,1血清二胺氧化酶活性、内毒素和D-乳酸含量都出现显著增加(P<0.05);2组成肠黏膜细胞间紧密连接结构的闭合蛋白Claudin-3、Claudin-15a和胞浆蛋白ZO-1、ZO-2、ZO-3基因表达活性显著下降(P<0.05),而闭锁蛋白Occludin基因表达活性出现不同程度的下降,但差异不显著(P>0.05);3透射电镜观察结果显示肠黏膜细胞间紧密连接结构出现缝隙;4饲料中的AV值、POV值、MDA含量与闭合蛋白Claudin-3、Claudin-15a,胞浆蛋白ZO-1、ZO-2、ZO-3和闭锁蛋白Occludin基因表达活性均显示负相关关系,其中饲料POV值、MDA含量与胞浆蛋白ZO-1基因表达活性呈极显著负相关关系(P<0.05)。结果表明,在饲料中添加氧化鱼油后,氧化鱼油中的过氧化物、丙二醛等油脂氧化产物导致组成肠道黏膜细胞间紧密连接结构的蛋白基因表达活性显著下降、紧密连接物理性结构显著损伤,致使肠黏膜屏障结构的完整性被破坏、肠黏膜通透性显著增加。
In this experiment, the effect of feed oxidized fish oil on intestinal barrier structure and intestinal permeability of grass carp was studied. Fish oil group (6F), soybean oil group (6S), 2% oxidized fish oil (2OF), 4% oxidized fish oil (4OF) and 6% oxidized fish oil (6OF) were designed with soybean oil, fish oil and oxidized fish oil as feed fat source Group and other nitrogen, semi-pure feed can be semi-purified, cage fish culture ponds [average body weight (74.8 ± 1.2) g] 72 d. The occludins Occludin, Claudin-3, Claudin-15a and ZO-1, ZO-2 and ZO-3 were determined by real-time quantitative PCR Of the gene expression activity, combined with intestinal mucosal tight junction structure of transmission electron microscopy, intestinal permeability markers of comprehensive analysis. The results showed that the activities of serum diamine oxidase, endotoxin and D-lactate increased significantly (P <0.05) after the addition of fish oil; 2 Claudin-3, a closed- The expression of Claudin-15a and ZO-1, ZO-2 and ZO-3 were significantly decreased (P <0.05), while the expression of occludin gene Occludin was decreased to some extent, but the difference was insignificant ); 3 Transmission electron microscopy showed that there was a gap in the tight junctions between intestinal mucosa cells; 4 The values of AV, POV and MDA in feedstuffs were significantly correlated with the expressions of Claudin-3, Claudin-15a, ZO- 2, ZO-3 and occludin Occludin gene expression showed a negative correlation, the feed POV value, MDA content and ZO-1 cytoplasmic protein gene expression activity was significantly negatively correlated (P <0.05). The results showed that oxidation of fish oil with peroxide and malondialdehyde resulted in a significant decrease in the expression of protein gene expression in the tight junctions between intestinal mucosa and the physical structure of tight junctions Damage, resulting in the integrity of the intestinal mucosal barrier structure is destroyed, significantly increased intestinal mucosal permeability.