动物营养学报 >

2020 , Vol. 32 >Issue 6: 2816 - 2823

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.06.042

分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

海藻多糖对仔猪小肠上皮细胞紧密连接蛋白及核转录因子-κB信号通路相关基因表达的影响

  • 郭晓波 ,
  • 邹田德 ,
  • 杨晋 ,
  • 曾永娣 ,
  • 王自蕊 ,
  • 游金明
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  • 1. 江西农业大学, 江西省动物营养重点实验室, 江西省营养饲料开发工程研究中心, 江西省优质安全畜禽生产产教融合重点创新中心, 南昌 330045;
    2. 赣州市畜牧研究所, 赣州 341401
郭晓波(1993-),女,河北张家口人,硕士研究生,研究方向为猪营养与饲料科学。E-mail:348310587@qq.com

收稿日期: 2020-01-07

  网络出版日期: 2020-06-16

基金资助

江西省生猪产业技术体系（JXARS-03-营养与饲料岗）

收起

Effects of Seaweed Polysaccharides on Expression of Tight Junction Protein and Related Genes in Nuclear Transcription Factor-κB Signaling Pathway in Porcine Epithelial Cells

  • GUO Xiaobo ,
  • ZOU Tiande ,
  • YANG Jin ,
  • ZENG Yongdi ,
  • WANG Zirui ,
  • YOU Jinming
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  • 1. Jiangxi Province Key Innovation Center for Industry-Education Integration of High Quality and Safety Livestock Production, Jiangxi Province Feed Development Engineering Research Center, Jiangxi Province Key Laboratory of Animal Nutrition, Jiangxi Agricultural University, Nanchang 330045, China;
    2. Ganzhou Animal Husbandry Research Institute, Ganzhou 341401, China

Received date: 2020-01-07

  Online published: 2020-06-16

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摘要

本试验旨在研究不同浓度海藻多糖对仔猪小肠上皮细胞（IPEC-J2细胞）紧密连接蛋白及核转录因子-κB （NF-κB）信号通路相关基因表达的影响。IPEC-J2细胞加入浓度分别为0（对照）、60、120和240 μg/mL的海藻多糖处理培养基培养24 h，分析海藻多糖对IPEC-J2细胞机械屏障和免疫屏障相关基因以及蛋白表达的影响。结果表明：1）与对照组相比，60、120 μg/mL的海藻多糖可以显著上调紧密连接蛋白闭锁蛋白（occludin）和闭合小环蛋白-1（ZO-1）的mRNA相对表达量（P<0.05）；但240 μg/mL海藻多糖组occludin和ZO-1的mRNA相对表达量与对照组无显著差异（P>0.05）。与对照组相比，120和240 μg/mL的海藻多糖可以显著上调封闭蛋白-1（claudin-1）的mRNA相对表达量（P<0.05）。与对照组相比，120、240 μg/mL的海藻多糖可以显著上调claudin-1和ZO-1的蛋白相对表达量（P<0.05）。2）与对照组相比，60、120和240 μg/mL的海藻多糖均显著降低了髓样分化蛋白88（MyD88）和p65的mRNA相对表达量（P<0.05），240 μg/mL的海藻多糖显著降低了Toll样受体-4（TLR-4）和核转录因子-κB抑制蛋白α（IκBα）的mRNA相对表达量（P<0.05）。由此可知，海藻多糖可以上调IPEC-J2细胞紧密连接蛋白occludin、claudin-1和ZO-1的mRNA相对表达量，降低IPEC-J2细胞NF-κB信号通路相关基因的表达，对改善仔猪小肠上皮细胞屏障功能具有积极的作用。

关键词： 海藻多糖; IPEC-J2; 肠道屏障; 紧密连接蛋白; NF-&kappa; B信号通路

本文引用格式

郭晓波 , 邹田德 , 杨晋 , 曾永娣 , 王自蕊 , 游金明 . 海藻多糖对仔猪小肠上皮细胞紧密连接蛋白及核转录因子-κB信号通路相关基因表达的影响[J]. 动物营养学报, 2020 , 32(6) : 2816 -2823 . DOI: 10.3969/j.issn.1006-267x.2020.06.042

Abstract

This study aimed to evaluate the effects of different concentration of seaweed polysaccharides on expression of tight junction protein and related genes in nuclear transcription factor-κB (NF-κB) signaling pathway in porcine epithelial cells (IPEC-J2 cells). The IPEC-J2 cells were treated with seaweed polysaccharides culture medium with concentrations of 0 (control), 60, 120 and 240 μg/mL, cultured for 24 h, and the effects of seaweed polysaccharides on expression of mechanical barrier and immune barrier related gene and protein in IPEC-J2 cells were analysed. The results showed as follows: 1) compared with the control group, 60 and 120 μg/mL seaweed polysaccharides significantly upregulated the mRNA relative expression levels of occludin and zonula occluden-1 (ZO-1) (P<0.05); but no significant differences were observed in the mRNA relative expression levels of occludin and ZO-1 between control group and 240 μg/mL seaweed polysaccharides group (P>0.05). Compared with control group, 120 and 240 μg/mL seaweed polysaccharides significantly upregulated the mRNA relative expression level of claudin-1 (P<0.05). Compared with control group, 120 and 240 μg/mL seaweed polysaccharides significantly upregulated the protein relative expression levels of claudin-1 and ZO-1 (P<0.05). 2) Compared with control group, 60, 120 and 240 μg/mL seaweed polysaccharides significantly decreased the mRNA relative expression levels of myeloid differentiation protein 88 (MyD88) and p65 (P<0.05), and 240 μg/mL seaweed polysaccharides significantly decreased the mRNA relative expression levels of Toll like receptor-4 (TLR-4) and nuclear transcription factor inhibitor protein α (IκBα) (P<0.05). In conclusion, seaweed polysaccharides can upregulate the mRNA relative expression levels of occludin, claudin-1 and ZO-1 in IPEC-J2 cells, and decrease the expression of NF-κB signaling pathway related gene in IPEC-J2 cells, and have positive effects on improv the barrier function of porcine epithelial cells.

Key words： seaweed polysaccharide; IPEC-J2; intestinal barrier; tight junction protein; NF-κB signaling pathway

参考文献

[1] 吴兆海.粪菌移植对被动免疫失败犊牛肠道屏障功能及肠道菌群构建的影响[D].博士学位论文.北京:中国农业大学,2018.
[2] 黄妍君.维生素E及其衍生物对肠道屏障结构蛋白的保护作用研究[D].硕士学位论文.长沙:湖南师范大学,2018:1-2.
[3] FANNING A S,MITIC L L,ANDERSON J M.Transmembrane proteins in the tight junction barrier[J].Journal of the American Society of Nephrology,1999,10(6):1337-1345.
[4] 刘畅.探究TLR4对小鼠肠黏膜紧密连接蛋白的调控作用[D].硕士学位论文.沈阳:中国医科大学,2018.
[5] 崔勇鹤,孙少明,李占武,等.肠道上皮细胞在肠道免疫屏障中的维护及炎性肠病发生机制[J].中国中西医结合消化杂志,2019,27(2):160-162.
[6] 夏溪.猪抗菌肽PR39抗细菌感染和保护肠道屏障功能的作用及其机制研究[D].博士学位论文.杭州:浙江大学,2015.
[7] 王凤杰.HIV-1感染致肠道上皮紧密连接异常的分子机制[D].博士学位论文.北京:中国疾病预防控制中心,2019:13-116.
[8] CUMASHI A,USHAKOVA N A,PREOBRAZHENSKAYA M E,et al.A comparative study of the anti-inflammatory,anticoagulant,antiangiogenic,and antiadhesive activities of nine different fucoidans from brown seaweeds[J].Glycobiology,2007,17(5):541-552.  
[9] WAN J,JIANG F,XU Q S,Alginic acid oligosaccharide accelerates weaned pig growth through regulating antioxidant capacity,immunity and intestinal development[J].RSC Advances,2016,6(90):87026-87035.
[10] HWANG P A,LIN H V,LIN H Y,et al.Dietary supplementation with low-molecular-weight fucoidan enhances innate and adaptive immune responses and protects against Mycoplasma pneumonia antigen stimulation[J].Marine Drugs,2019,17(3):175.
[11] CIAN R E,DRAGO S R,DE MEDINA F S,et al.Proteins and carbohydrates from red seaweeds:evidence for beneficial effects on gut function and microbiota[J].Marine Drugs,2015,13(8):5358-5383.  
[12] CHOI Y S,EOM S Y,KIM I S,et al.Fucoidan extracted from hijiki protects brain microvessel endothelial cells against diesel exhaust particle exposure-induced disruption[J].Journal of Medicinal Food,2016,19(5):466-471.  
[13] 赵省如.决明子对小鼠肠道屏障影响的研究[D].硕士学位论文.南昌:南昌大学,2017.
[14] 刘珂,邱炳勋,邹利,等.黄芪及其有效成分对内皮细胞及其连接的保护作用及机制研究进展[J].中草药,2016,47(21):3912-3917.
[15] 谢红兵,邹云,刘丽莉,等.植物多糖对断奶仔猪小肠黏膜形态及肠黏膜屏障功能的影响[J].中国兽医学报,2019,39(1):150-157,187.
[16] HAN J,LI J,BAI G,et al.Acanthopanax senticosus polysaccharides-induced intestinal tight junction injury alleviation via inhibition of NF-κB/MLCK pathway in a mouse endotoxemia model[J].World Journal of Gastroenterology,2017,23(12):2175-2184.  
[17] WERTZ I E,DIXIT V M.Signaling to NF-κB:regulation by ubiquitination[J].Cold Spring Harbor Perspectives in Biology,2010,2(3):a003350.
[18] 游金明,郭晓波,王自蕊,等.海藻多糖的免疫调节作用及其在畜禽生产中的应用[J].饲料工业,2017,38(20):1-5.
[19] HEIM G,O'DOHERTY J V,O'SHEA C J,et al.Maternal supplementation of seaweed-derived polysaccharides improves intestinal health and immune status of suckling piglets[J].Journal of Nutritional Science,2015,4:e27.
[20] JEONG S Y,SEOL D W.The role of mitochondria in apoptosis[J].BMB Reports,2008,41(1):11-22.  
[21] AISA Y,MIYAKAWA Y,NAKAZATO T,et al.Fucoidan induces apoptosis of human HS-Sultan cells accompanied by activation of caspase-3 and down-regulation of ERK pathways[J].American Journal of Hematology,2005,78(1):7-14.  
[22] NEELAKANDAN Y,VENKATESAN A.Antinociceptive and anti-inflammatory effect of sulfated polysaccharide fractions from Sargassum wightii and Halophila ovalis in male Wistar rats[J].Indian Journal of Pharmacology,2016,48(5):562-570.  
[23] WALSH A M,SWEENEY T,O'SHEA C J,et al.Effects of supplementing dietary laminarin and fucoidan on intestinal morphology and the immune gene expression in the weaned pig[J].Journal of Animal Science,2012,90(Suppl.4):284-286.
[24] BAHAR B,O'DOHERTY J V,HAYES M,et al.Extracts of brown seaweeds can attenuate the bacterial lipopolysaccharide-induced pro-inflammatory response in the porcine colon ex vivo[J].Journal of Animal Science,2012,90(Suppl.4):46-48.
[25] SANJEEWA K K A,JAYAWARDENA T U,KIM H S,et al.Fucoidan isolated from Padina commersonii inhibit LPS-induced inflammation in macrophages blocking TLR/NF-κB signal pathway[J].Carbohydrate Polymers,2019,224:115195.
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