Effects of Lipopolysaccharide Challenge on Key Genes Expression of Inflammatory Signaling Pathway in Spleen of Weaned Piglets

  • WANG Yang ,
  • LI Xiangen ,
  • LIU Yulan ,
  • ZHU Huiling ,
  • WANG Xiuying
Expand
  • Hubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic University, Wuhan 430023, China

Received date: 2019-01-22

  Online published: 2019-08-19

Abstract

The purpose of this study was to investigate the changes of key genes of Toll-like receptor 4 (TLR4) and nucleotide-bound oligomeric domain receptor (NOD) inflammation signaling pathway in spleen of weaned piglets at different time after lipopolysaccharide (LPS) challenge. Forty-two Duroc×Landrace×Yorkshine weaned piglets with a body weight of (7.1±0.9) kg were selected and randomly divided into 7 treatments (6 pigs per treatment). The piglets were injected with 100 μg/kg body weight LPS, and slaughtered at 0 h (before LPS challenge), 1, 2, 4, 8, 12 and 24 h (after LPS challenge). Blood and spleen samples were taken to determine the blood cell differential counts and the mRNA expressions of inflammatory related genes, respectively. The results showed as follows:LPS challenge led to significant changes in blood cell classification count (P<0.05). The mRNA expressions of key genes of TLR4 and NOD signaling pathway were increased after LPS challenge (P<0.05), and reached the peak at 2 to 8 h. The mRNA expressions of tumor necrosis factor-α, interleukin-1β, interleukin-6, cyclooxygenase 2 and heat shock protein 70 were also significantly increased after LPS challenge (P<0.05), and peaked at 1 to 4 h. At 8 to 24 h post injection, these genes gradually returned to normal level or slightly lower than normal level. Overall, LPS challenge activates the inflammatory signaling pathway and up-regulates the expression of inflammatory cytokines. During LPS stimulation, the expressions of these gene increase at the early time, and then decrease gradually after the peak.

Cite this article

WANG Yang , LI Xiangen , LIU Yulan , ZHU Huiling , WANG Xiuying . Effects of Lipopolysaccharide Challenge on Key Genes Expression of Inflammatory Signaling Pathway in Spleen of Weaned Piglets[J]. Chinese Journal of Animal Nutrition, 2019 , 31(8) : 3882 -3890 . DOI: 10.3969/j.issn.1006-267x.2019.08.050

References

[1] LEE I K,KEY Y C,KIM G,et al.Stress,nutrition,and intestinal immune responses in pigs-a review[J].Asian-Australasian Journal of Animal Sciences,2016,29(8):1075-1082.  
[2] ZHANG L,WANG X Y,CHEN S K,et al.Medium-chain triglycerides attenuate liver injury in lipopolysaccharide-challenged pigs by inhibiting necroptotic and inflammatory signaling pathways[J].International Journal of Molecular Sciences,2018,19(11):3697.
[3] ZHU H L,WANG H B,WANG S H,et al.Flaxseed oil attenuates intestinal damage and inflammation by regulating necroptosis and TLR4/NOD signaling pathways following lipopolysaccharide challenge in a piglet model[J].Molecular Nutrition & Food Research,2018,62(9):e1700814.
[4] 李宗芳,张澍.脾脏的基础研究进展与展望[J].西安交通大学学报(医学版),2008,29(1):1-6.
[5] WANG L M,TU Z X,WANG H B,et al.Flaxseed oil improves liver injury and inhibits necroptotic and inflammatory signaling pathways following lipopolysaccharide challenge in a piglet model[J].Journal of Functional Foods,2018,46:482-489.
[6] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and 2-△△CT method[J].Methods,2001,25(4):402-408.  
[7] 张澍,李宗芳.脾脏功能与脾脏外科研究现状与展望[J].中华实验外科杂志,2014,31(2):231-233.
[8] 张华,侯绍华.血常规检查在犬猫疾病诊断中的应用[J].中国动物检疫,2009,26(3):55-56.
[9] 汪洋,王秀英,汪龙梅,等.Necrostatin-1对脂多糖刺激的断奶仔猪血细胞分类计数和血液生化指标的影响[J].中国畜牧杂志,2018,54(8):80-86.
[10] 郭峰.红细胞免疫的研究和意义[J].自然杂志,2002,24(5):268-273.
[11] GROS A,OLLIVIER V,HO-TIN-NOÉ B.Platelets in inflammation:regulation of leukocyte activities and vascular repair[J].Frontiers in Immunology,2015,5:678.
[12] 陈维佩.脾脏解剖生理及外科手术的研究进展[J].局解手术学杂志,1994(3):34-37.
[13] WU H T,LIU Y L,PI D A,et al.Asparagine attenuates hepatic injury caused by lipopolysaccharide in weaned piglets associated with modulation of toll-like receptor 4 and nucleotide-binding oligomerisation domain protein signalling and their negative regulators[J].British Journal of Nutrition,2015,114(2):189-201.  
[14] 王秀英.谷氨酸对脂多糖诱导的仔猪肠道损伤及肌肉蛋白质合成和降解的调控作用[D].硕士学位论文.武汉:武汉轻工大学,2015.
[15] WANG X Y,LIU Y L,WANG S H,et al.Asparagine reduces the mRNA expression of muscle atrophy markers via regulating protein kinase B (Akt),AMP-activated protein kinase α,toll-like receptor 4 and nucleotide-binding oligomerisation domain protein signalling in weaning piglets after lipopolysaccharide challenge[J].British Journal of Nutrition,2016,116(7):1188-1198.  
[16] TAKAHASHI Y,ISUZUGAWA K,MURMSE Y,et al.Up-regulation of NOD1 and NOD2 through TLR4 and TNF-α in LPS-treated murine macrophages[J].Journal of Veterinary Medical Science,2006,68(5):471-478.  
[17] LIU Z Y,WU B,GUO Y S,et al.Necrostatin-1 reduces intestinal inflammation and colitis-associated tumorigenesis in mice[J].American Journal of Cancer Research,2015,5(10):3174-3185.
[18] SAINI R K,SANYAL S N,BHATTI J S.Chemopreventive action of non-steroidal anti-inflammatory drugs in 9,10-dimethylbenzanthracene induced lung carcinogenesis in BALB/C mice:expression of COX-1,COX-2 and Nf-κB[J].Journal of Applied Biomedicine,2018,16(4):320-327.  
[19] LANG C H,SILVIS C,DESHPANDE N,et al.Endotoxin stimulates in vivo expression of inflammatory cytokines tumor necrosis factor α,interleukin-1β,-6,and high-mobility-group protein-1 in skeletal muscle[J].Shock,2003,19(6):538-546.  
[20] ROSENSTIEL P,FANTINI M,BRÄUTIGAM K,et al.TNF-α and IFN-γ regulate the expression of the NOD2 (CARD15) gene in human intestinal epithelial cells[J].Gastroenterology,2003,124(4):1001-1009.  
[21] 谢翔,应炜阳,金胜威.COX-2在炎症消退中的研究进展[J].生命的化学,2016,36(4):461-464.
[22] 刘嘉敏,赵元莙.HSP70的研究进展及其在生物医学中的应用[J].教育教学论坛,2017(50):61-62.
[23] KUBOKI S,SCHUSTER R,BLANCHARD J,et al.Role of heat shock protein 70 in hepatic ischemia-reperfusion injury in mice[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2007,292(4):G1141-G1149.
Outlines

/