单 位：
吉林农业大学吉林省动物营养与饲料科学重点实验室
关键词：
刺五加苷;仔猪;小肠上皮细胞;紧密连接蛋白;细胞因子
摘 要：
为研究刺五加苷E(Eleutheroside E)对仔猪小肠上皮(IPEC-J2)细胞紧密连接蛋白及炎性细胞因子m RNA表达量的影响。在正常DMEM/F12培养基中分别添加0,0. 05,0. 1,0. 2 mg/m L的刺五加苷E,测定刺五加苷E对IPEC-J2细胞增殖活性的影响。选择对IPEC-J2有显著作用的刺五加苷E浓度为有效浓度,研究刺五加苷E对仔猪小肠上皮细胞的机械屏障和免疫屏障的影响。细胞增殖试验结果表明,0. 1 mg/m L的刺五加苷E对IPEC-J2有极显著的增殖作用(P<0. 01)。选择0. 1 mg/m L的刺五加苷E处理剂量对IPEC-J2的主要紧密连接蛋白和细胞因子进行基因表达水平测定,采用实时荧光定量PCR方法测定紧密连接Claudin-3、Occludin和ZO-1,抗炎性细胞因子IL-10和TGF-β,炎性细胞因子IL-6、TNF-α、INF-γ的m RNA相对表达量。试验结果表明:刺五加苷E上调了仔猪小肠上皮细胞紧密连接蛋白Claudin-3和ZO-1的基因表达(P<0. 05),在细胞免疫方面下调炎性细胞因子IL-6、TNF-α、INF-γ的基因表达;上调了抗炎性细胞因子IL-10和TGF-β的基因表达。这说明刺五加苷E能够改变细胞间紧密连接蛋白和细胞因子的基因表达,进而促进肠道机械和免疫屏障功能的完整性,有益于仔猪肠道的健康发育。
译 名：
Effects of Eleutheroside E on Expression Level of Barrier Function Gene in Small Intestinal Epithelial Cells of Piglets
作 者：
FAN Yueli;ZHAO Bao;CHE Dongsheng;Jilin Provincial Key Laboratory of Animal Nutrition and Feed Science,Jilin Agricultural University;College of Animal Science and Technology,Jilin Agricultural University;Key Laboratory of Animal Production,Product Quality and Security,Ministry of Education,Jilin Agricultural University;
关键词：
Eleutheroside;;piglet;;small intestinal epithelial cell;;tight junction protein;;cytokine
摘 要：
The aim of the study was to investigate the effects of Eleutheroside E(EE) on the expression of tight junction protein and inflammatory cytokines(m RNA) in IPEC-J2 cells of the small intestinal epithelium of piglets. 0 mg/m L,0. 05 mg/m L,0. 1 mg/m L and 0. 2 mg/m L of EE were respectively in normal DMEM/F12 medium to determine the effects of EE on proliferation activity of IPEC-J2 cells. EE concentration with a significant effect on IPEC-J2 was selected as effective concentration,and EE effects on mechanical barrier and immune barrier of small intestinal epithelial cells were studied. The results of cell proliferation experiments showed that the EE at 0. 1 mg/m L had a significant proliferative effect on IPEC-J2(P<0. 01). The gene expression level of major tight junction proteins and cytokines of IPEC-J2 was determined by 0. 1 mg/m L EE treatment. Real-time fluorescence quantitative PCR was used to determine m RNA relative expression level of Claudin-3,Occludin and ZO-1,anti-inflammatory cytokines IL-10 and TGF-β,inflammatory cytokines IL-6,TNF-α and INF-γ. The results of vitro experiments showed that EE could up-regulate the expression of tight junction proteins Claudin-3 and ZO-1 in small intestinal epithelium of piglets(P < 0. 05),and down-regulated inflammatory cytokines IL-6,TNF-α and INF-γ gene expression; up-regulated the expression of anti-inflammatory cytokines IL-10 and TGF-β. These results suggested that EE could change the gene expression of intercellular tight junction proteins and cytokines,and promote the integrity of intestinal barrier function. Therefore,EE might be beneficial to health development of intestinal tract of piglets.
