动物营养学报 >

2012 , Vol. 24 >Issue 7: 1384 - 1388

DOI: https://doi.org/10.3969/j.issn.1006-267x.2012.07.025

研究简报

长链n-3多不饱和脂肪酸对肠上皮细胞促炎细胞因子基因mRNA表达的影响

  • 黄志清 ,
  • 陈小玲 ,
  • 陈代文 ,
  • 余冰 ,
  • 何军
展开
  • 四川农业大学动物营养研究所, 动物抗病营养教育部重点实验室, 雅安 625014

收稿日期: 2012-03-22

  网络出版日期: 2012-07-05

基金资助

四川省教育厅自然科学重点项目(10ZA044)

收起

Long-Chain n-3 Polyunsaturated Fatty Acids Affect mRNA Expression of Pro-Inflammatory Cytokine Genes in Intestinal Epithelial Cells

  • HUANG Zhiqing ,
  • CHEN Xiaoling ,
  • CHEN Daiwen ,
  • YU Bing ,
  • HE Jun
Expand
  • Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Ya’an 625014, China

Received date: 2012-03-22

  Online published: 2012-07-05

Fold

摘要

本试验旨在探讨长链n-3多不饱和脂肪酸(LC n-3 PUFA)对肠上皮细胞促炎细胞因子基因mRNA表达的影响。试验选用大鼠肠上皮细胞系IEC-6细胞为模型,分为4个处理,分别为对照、脂多糖(LPS,1 μg/mL)、LPS(1 μg/mL)+二十二碳六烯酸(DHA,100 μmol/L)和LPS(1 μg/mL)+二十碳五烯酸(EPA,100 μmol/L),每个处理3个重复,每孔为1个重复。细胞先用DHA、EPA或等量二甲基亚砜(DHA和EPA的溶剂,对照)预处理48 h,再用LPS处理3 h,收集细胞提取总RNA,采用实时定量PCR方法分析肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)和白介素-6(IL-6)的基因mRNA表达水平的差异。结果表明:LPS极显著上调了细胞中TNF-α、IL-1β和IL-6的基因mRNA表达水平(P<0.01),EPA均极显著或显著削弱了细胞内LPS诱导的TNF-α(P<0.01)、IL-1β(P<0.01)和IL-6的基因mRNA水平(P<0.05)的上调,而DHA仅显著削弱了细胞内LPS诱导的IL-1β的基因mRNA水平的上调(P<0.05)。结果提示,LC n-3 PUFA在肠上皮细胞中具有抗炎作用,且在本试验条件下EPA的抗炎效果要优于DHA。

关键词： 长链n-3多不饱和脂肪酸; 二十碳五烯酸; 二十二碳六烯酸; IEC-6细胞; 促炎细胞因子

本文引用格式

黄志清 , 陈小玲 , 陈代文 , 余冰 , 何军 . 长链n-3多不饱和脂肪酸对肠上皮细胞促炎细胞因子基因mRNA表达的影响[J]. 动物营养学报, 2012 , 24(7) : 1384 -1388 . DOI: 10.3969/j.issn.1006-267x.2012.07.025

Abstract

This present study was conducted to investigate the effects of long-chain n-3 polyunsaturated fatty acids (LC n-3 PUFA) on the mRNA expression of pro-inflammatory cytokine genes in intestinal epithelial cells. IEC-6 cells from rat intestinal epithelial cells were used as the experimental model and subjected to four treatments with three replicates in each treatment and one well per replicate. The treatments were as follows: control, lipopolysaccharide (LPS, 1 μg/mL), LPS(1 μg/mL)+docosahexaenoic acid (DHA, 100 μmol/L) and LPS (1 μg/mL)+eicosapentaenoic acid (EPA,100 μmol/L). IEC-6 cells was collected for total RNA extraction after 48 h pretreatment with DHA, EPA or equivalent amounts of dimethyl sulfoxide (solvent of DHA and EPA, control) and 3 h treatment with LPS. The mRNA expression levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) genes were determined using real-time quantitative PCR. The results showed that the mRNA expression levels of TNF-α, IL-1β and IL-6 genes were significantly up-regulated after LPS challenge (P<0.01), EPA significantly attenuated LPS-induced upregulation of mRNA expression levels of TNF-α (P<0.01), IL-1β (P<0.01) and IL-6 genes (P<0.05), while DHA only significantly attenuated LPS-induced upregulation of mRNA expression level of IL-1β gene (P<0.05). These results indicate that LC n-3 PUFA has an anti-inflammatory effect in intestinal epithelial cells, and the effect of EPA is more efficient than that of DHA under conditions in this experiment.

Key words： long-chain n-3 polyunsaturated fatty acid; eicosapentaenoic acid; docosahexaenoic acid; IEC-6 cells; pro-inflammatory cytokines

参考文献

[1] MAYER K,MEYER S,REINHOLZ M M,et al.Short-time infusion of fish oil based lipid emulsions,approved for parenteral nutrition,reduces monocyte proinflammatory cytokine generation and adhesive interaction with endothelium in humans[J].Journal of Immunology,2003,171(9):4837-4843.
[2] WELDON S M,MULLEN A C,LOSCHER C E,et al.Docosahexaenoic acid induces an anti-inflammatory profile in lipopolysaccharide-stimulated human THP-1 macrophages more effectively than eicosapentaenoic acid[J].Journal of Nutritional Biochemistry,2007,18(4):250-258.  
[3] MARCHESELLI V L,HONG S,LUKIW W J,et al.Novel docosanoids inhibit brain ischemia-reperfusion-mediated leukocyte infiltration and pro-inflammatory gene expression[J].Journal of Biological Chemistry,2003,278(44):43807-43817.  
[4] MOON D O,KIM K C,JIN C Y,et al.Inhibitory effects of eicosapentaenoic acid on lipopolysaccharide-induced activation in BV2 microglia[J].International Immunopharmacology,2007,7(2):222-229.  
[5] SAEDISOMEOLIA A,WOOD L G,GARG M L,et al.Anti-inflammatory effects of long-chain n-3 PUFA in rhinovirus-infected cultured airway epithelial cells[J].British Journal of Nutrition,2009,101(4):533-540.  
[6] 史艳莉,李翠娥,代方梅,等.n-3多不饱和脂肪酸对运动性哮喘豚鼠气道炎症和气道高反应性的影响[J].中国运动医学杂志,2010,29(6):669-672,687.
[7] LIU F,LI G,WEN K,et al.Porcine small intestinal epithelial cell line (IPEC-J2) of rotavirus infection as a new model for the study of innate immune responses to rotaviruses and probiotics[J].Viral Immunology,2010,23(2):135-149.  
[8] PFAFFL M W.A new mathematical model for relative quantification in real-time RT-PCR[J].Nucleic Acids Research,2001,29:45.
[9] QUARONI A,WANDS J,TRELSTAD R L,et al.Epithelioid cell cultures from rat small intestine:characterization by morphologic and immunological criteria[J].Journal of Cell Biology,1979,80(2):248-265.  
[10] ARCE C,RAMÍREZ-BOO M,LUCENA C,et al.Innate immune activation of swine intestinal epithelial cell lines (IPEC-J2 and IPI-2I) in response to LPS from Salmonella typhimurium[J].Comparative Immunology Microbiology and Infectious Diseases,2010,33(2):161-174.  
[11] LYU S Y,PARK W B.Mistletoe lectin modulates intestinal epithelial cell-derived cytokines and B cell IgA secretion[J].Archives of Pharmacal Research,2009,32(3):443-451.  
[12] 蔡景义,周安国,田刚,等.氧化锌对LPS处理猪肠上皮细胞炎性细胞因子分泌及基因表达的影响[J].中国畜牧杂志,2011,47(23):47-50.
[13] CAUGHEY G E,MANTZIORIS E,GIBSON R A,et al.The effect on human tumor necrosis factor α and interleukin 1β production of diets enriched in n-3 fatty acids from vegetable oil or fish oil[J].American Journal of Clinical Nutrition,1996,63(1):116-122.
[14] CLELAND L G,JAMES M J,PROUDMAN S M.Dietary fats and inflammation:the medicinal use of fish oil[J].Nutrition & Dietetics,2009,66(1):4-6.  
[15] HALADE G V,WILLIAMS P J,LINDSEY M L,et al.Fish oil decreases inflammation and reduces cardiac remodeling in rosiglitazone treated aging mice[J].Pharmacological Research,2011,63(4):300-307.  
[16] MONK J M,KIM W,CALLAWAY E,et al.Immunomodulatory action of dietary fish oil and targeted deletion of intestinal epithelial cell PPAR delta in inflammation-induced colon carcinogenesis[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2012,302(1):G153-G167.
[17] KEW S,MESA M D,TRICON S,et al.Effects of oils rich in eicosapentaenoic and docosahexaenoic acids on immune cell composition and function in healthy humans[J].American Journal of Clinical Nutrition,2004,79(4):674-681.
[18] ALEXANDER J W.Immunonutrition:the role of N-3 fatty acids[J].Nutrition,1998,14(7/8):627-633.
Options
文章导航

/

〈 〉

    联系我们

  • 地址：北京市海淀区圆明园西路2号
    　　　中国农业大学动科动医大楼153室
  • 邮编：100193
  • 电话：010－62817823
  • 网址：http://www.chinajan.com
  • E-mail：cjan@chinajan.com
京公网安备 11010802041926号    京ICP备17050989号-2