摘要
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。
A 72- day experiment was carried out to investigate the effect of oxidized fish oil on intestinal mucosa barrier structure and intestinal permeability of grass carp（ Ctenopharyngodon idella） [average weight（ 74. 8 ± 1. 2） g]. Five diets containing different levels of soybean oil,fish oil and oxidized fish oil（ 6% fish oil,6% soybean oil,2% oxidized fish oil; 4% oxidized fish oil,6% oxidized fish oil） were evaluated according to the equal standards of nitrogen and energy. Using qRT- PCR method,the gene expression levels about atresia protein Occludin,closed protein Claudin3,Claudin15 a and cytoplasmic protein ZO- 1,ZO- 2,ZO- 3 in the intestine were measured,and TEMobservations of intestinal mucosal cells tight junction and intestinal permeability index were comprehensively analyzed. The results showed that,（ 1）Serum diamine oxidase activity,endotoxin and D-lactic acid content significantly increased under oxidized fish oil（ P〈0. 05）.（ 2） The gene expression of closed protein Claudin-3,Claudin-15 a and cytoplasmic protein ZO- 1,ZO- 2,ZO- 3 were significantly decreased under oxidized fish oil（ P〈0. 05）.（ 3） The tight junction structure of intestinal mucosal cells displayed gap under oxidized fish oil.（ 4） The values of AV,POV and the content of MDA in diet downregulated with closed protein Claudin3,Claudin15 a and cytoplasmic protein ZO- 1,ZO- 2,ZO- 3 gene expression,besides,the value of POV and the content of MDA in diet significantly downregulated with cytoplasmic protein ZO- 1 gene expression under oxidized fish oil（ P〈0. 05）. Collectively,the present study demonstrated that after adding oxidized fish oil in feed,peroxide oxidation and other oil oxidation product like malondialdehyde of fish oil led to the fact that tight junction protein gene expression activity significantly decreased and tight junction physical structure significant damaged, resulting in the fact that intestinal barrier structure was destroyed and intestinal permeability significantly increased.
出处
《水产学报》
CAS
CSCD
北大核心
2015年第10期1511-1520,共10页
Journal of Fisheries of China
基金
国家自然科学基金(31172417)
苏州市应用基础研究项目(SYN201316)
关键词
氧化鱼油
草鱼
肠道
细胞紧密连接
通透性
oxidized fish oil
grass carp
intestine
cell tight junction
permeability
