Effects of Lipopolysaccharide on Inflammation and Protein Degradation-Related Genes and miRNA Expression in Gastrocnemius of Piglets after 24 h Stimulation

FAN Jiajun; HUANG Xingfa; WANG Yang; LI Xiangen; WAN Zhicheng; LIU Yulan; KANG Ping

doi:10.3969/j.issn.1006-267x.2021.10.039

2021 , Vol. 33 >Issue 10: 5802 - 5808

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.10.039

MOLECULAR AND CELLULAR NUTRITION

Effects of Lipopolysaccharide on Inflammation and Protein Degradation-Related Genes and miRNA Expression in Gastrocnemius of Piglets after 24 h Stimulation

FAN Jiajun ,
HUANG Xingfa ,
WANG Yang ,
LI Xiangen ,
WAN Zhicheng ,
LIU Yulan ,
KANG Ping

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Hubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic University, Wuhan 430023, China

Received date: 2021-03-30

Online published: 2021-10-16

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Abstract

This experiment was conducted to investigate the expression changes of genes and miRNA related to inflammation and protein degradation in gastrocnemius of weaned piglets challenged by lipopolysaccharide (LPS) at 24 h. Twelve healthy Duroc×Landrace×Yorkshine 28-day-old weaned piglets[body weight:(7.09±0.90) kg] were selected, and randomly divided into 2 groups:control group (n=6) and LPS group (n=6). All piglets were allowed ad libitum access to water and feed. On day 15 of experiment, the piglets in LPS group were injected with LPS (100 μg/kg BW) and sacrificed at 24 h post-injection. The control group received the same amount of sterile saline, and sacrificed immediately after the injection. Gastrocnemius samples were selected for testing. The results showed as follows:at 24 h after LPS treatment, the mRNA expression levels of inflammatory factors interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), as well as the key genes of Toll-like receptor 4 (TLR4) signaling pathway including TLR4, myeloid differentiation factor 88 (MyD88) and interleukin receptor-associated kinase 1 (IRAK1) were significantly increased in gastrocnemius (P<0.05), and the mRNA expression levels of nuclear factor-κB (NF-κB) and protein degradation-related gene forkhead box O4 (FOXO4) were also significantly increased (P<0.05). Moreover, miR-145 expression level in gastrocnemius was significantly increased (P<0.05), and the expression levels of miR-30b, miR-155, miR-132, miR-130a and miR-146a were significantly decreased at 24 h after LPS treatment (P<0.05). The results indicate that the gastrocnemius injury can be caused by the up-regulation of FOXO4 expression mediated by TLR4 signaling pathway at 24 h after LPS treatment. The miRNA may be involved in muscle inflammation and protein degradation at 24 h after LPS stimulation.

Key words： lipopolysaccharide; gastrocnemius; weaned piglet; inflammation; protein degradation; miRNA

Cite this article

FAN Jiajun , HUANG Xingfa , WANG Yang , LI Xiangen , WAN Zhicheng , LIU Yulan , KANG Ping . Effects of Lipopolysaccharide on Inflammation and Protein Degradation-Related Genes and miRNA Expression in Gastrocnemius of Piglets after 24 h Stimulation[J]. Chinese Journal of Animal Nutrition, 2021 , 33(10) : 5802 -5808 . DOI: 10.3969/j.issn.1006-267x.2021.10.039

References

[1] HUANG J R, ZHU X P.The molecular mechanisms of calpains action on skeletal muscle atrophy[J].Physiological Research, 2016, 65(4):547-560.
[2] 陈逢.鱼油通过TLR4和NOD信号通路对脂多糖诱导的仔猪肠道、肝脏损伤和肌肉蛋白质降解的调控作用[D].硕士学位论文.武汉:武汉轻工大学, 2013. CHEN F.Regulation of fish oil on intestinal and liver injury and muscle protein degradation induced by lipopolysaccharide in piglets through TLR4 and NOD signaling pathways[D].Master's Thesis.Wuhan:Wuhan Polytechnic University, 2013.(in Chinese)
[3] 汪洋.程序性坏死信号通路在脂多糖诱导的仔猪肠道损伤中的作用[D].硕士学位论文.武汉:武汉轻工大学, 2019. WANG Y.Role of programmed necrosis signaling pathway in lipopolysaccharide-induced intestinal injury in piglets[D].Master's Thesis.Wuhan:Wuhan Polytechnic University, 2019.(in Chinese)
[4] MCKERNAN D P.Pattern recognition receptors as potential drug targets in inflammatory disorder[J].Advances in Protein Chemistry and Structural Biology, 2020, 119:65-109.
[5] VAN QUICKELBERGHE E, DE SUTTER D, VAN LOO G, et al.A protein-protein interaction map of the TNF-induced NF-κB signal transduction pathway[J].Scientific data, 2018, 5:180289.
[6] ZHONG C, PU L Y, FANG M M, et al.ATRA regulates innate immunity in liver ischemia/reperfusion injury via RARα/Akt/Foxo1 signaling[J].Biological & Pharmaceutical Bulletin, 2018, 41(4):530-535.

[7] UWADA J, YAZAWA T, NAKAZAWA H, et al.Store-operated calcium entry (SOCE) contributes to phosphorylation of p38 MAPK and suppression of TNF-α signalling in the intestinal epithelial cells[J].Cellular Signalling, 2019, 63:109358.
[8] BARTEL D P.MicroRNAs:genomics, biogenesis, mechanism, and function[J].Cell, 2004, 116(2):281-297.

[9] WANG J, YANG L Z, ZHANG J S, et al.Effects of microRNAs on skeletal muscle development[J].Gene, 2018, 668:107-113.
[10] BRENNAN G P, HENSHALL D C.MicroRNAs as regulators of brain function and targets for treatment of epilepsy[J].Nature Reviews Neurology, 2020, 16(9):506-519.

[11] YADAV A, DAHUJA A, DABUR R.Dynamics of Toll-like receptors signaling in skeletal muscle atrophy[J/OL].Current Medicinal Chemistry, 2021.(2021-02-01).https://pubmed.ncbi.nlm.nih.gov/33530901/.doi:10.2174/0929867328666210202113734.
[12] BARREIRO E, PUIG-VILANOVA E, MARIN-CORRAL J, et al.Therapeutic approaches in mitochondrial dysfunction, proteolysis, and structural alterations of diaphragm and gastrocnemius in rats with chronic heart failure[J].Journal of Cellular Physiology, 2016, 231(7):1495-1513.

[13] NRC.Nutrient requirements of swine[S].10th ed.Washington, D.C.:The National Academies Press, 1998.
[14] 陈逢, 刘玉兰, 李权, 等.TLR4信号通路关键基因在断奶仔猪不同组织中的mRNA表达[J].中国畜牧杂志, 2013, 49(17):53-57. CHEN F, LIU Y L, LI Q, et al.The mRNA expression of key genes in TLR4 signaling pathway in different tissues of weaned piglets[J].Chinese Journal of Animal Science, 2013, 49(17):53-57.(in Chinese)
[15] LIVAK K J, SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J].Methods, 2001, 25(4):402-408.

[16] LIU Y L, HUANG J J, HOU Y Q, et al.Dietary arginine supplementation alleviates intestinal mucosal disruption induced by Escherichia coli lipopolysaccharide in weaned pigs[J].The British Journal of Nutrition, 2008, 100(3):552-560.

[17] ZHAO Y, MA D, DONG B C, et al.Per2 attenuates LPS-induced chondrocyte injury through the PTEN/PI3K/Akt signalling pathway[J].Bioscience Reports, 2020, 40(5):BSR20200417.
[18] KHAN H U, AAMIR K, JUSUF P R, et al.Lauric acid ameliorates lipopolysaccharide (LPS)-induced liver inflammation by mediating TLR4/MyD88 pathway in Sprague Dawley (SD) rats[J].Life Sciences, 2021, 265:118750.
[19] 王胜军, 刘妍, 刘玉兰, 等.炎症反应致肌肉蛋白质降解机理的研究进展[J].中国畜牧兽医, 2016, 43(9):2379-2387. WANG S J, LIU Y, LIU Y L, et al.Research progress on the mechanisms of inflammation-induced muscle protein degradation[J].China Animal Husbandry & Veterinary Medicine, 2016, 43(9):2379-2387.(in Chinese)
[20] 万志成, 汪龙梅, 张琳, 等.香菇多糖对脂多糖刺激断奶仔猪肌肉组织炎症反应及蛋白质降解相关基因表达的影响[J].动物营养学报, 2020, 32(3):1327-1333. WAN Z C, WANG L M, ZHANG L, et al.Effects of lentinan on expression of inflammation and protein degradation-related genes in muscle of weaned piglets challenged by lipopolysaccharide[J].Chinese Journal of Animal Nutrition, 2020, 32(3):1327-1333.(in Chinese)
[21] 李先根, 汪洋, 王秀英, 等.脂多糖刺激对断奶仔猪肌肉炎症和蛋白质降解相关基因表达的影响[J].中国畜牧杂志, 2019, 55(4):74-77. LI X G, WANG Y, WANG X Y, et al.Effects of lipopolysaccharide challenge on genes related to muscle inflammation and protein degradation of weaned piglets[J].Chinese Journal of Animal Science, 2019, 55(4):74-77.(in Chinese)
[22] KANG P, HUANG X F, WAN Z C, et al.Kinetics of changes in gene and microRNA expression related with muscle inflammation and protein degradation following LPS-challenge in weaned piglets[J].Innate Immunity, 2021, 27(1):23-30.

[23] WU H, FAN H, SHOU Z X, et al.Extracellular vesicles containing miR-146a attenuate experimental colitis by targeting TRAF6 and IRAK1[J].International Immunopharmacology, 2019, 68:204-212.
[24] FORDHAM J B, NAQVI A R, NARES S.Regulation of miR-24, miR-30b, and miR-142-3p during macrophage and dendritic cell differentiation potentiates innate immunity[J].Journal of Leukocyte Biology, 2015, 98(2):195-207.

[25] LI Y, YI M, WANG D, et al.LncRNA KCNQ1OT1 regulates endoplasmic reticulum stress to affect cerebral ischemia-reperfusion injury through targeting miR-30b/GRP78[J].Inflammation, 2020, 43(6):2264-2275.

[26] 张圆芳.基于miR-146a、miR-155调控NF-κB信号通路探讨芍药甘草汤治疗颈型颈椎病兔颈肌炎症损伤的作用机制[D].硕士学位论文.福州:福建中医药大学, 2017. ZHANG Y F.Mechanism of Shaoyao Gancao decoction on inflammatory injury of cervical muscle in rabbits with cervical spondylosis based on miR-146a and miR-155 regulating NF-κB signaling pathway[D].Master's Thesis.Fuzhou:Fujian University of Traditional Chinese Medical, 2017.(in Chinese)
[27] TILI E, MICHAILLE J J, CIMINO A, et al.Modulation of miR-155 and miR-125b levels following lipopolysaccharide/TNF-alpha stimulation and their possible roles in regulating the response to endotoxin shock[J].Journal of Immunology, 2007, 179(8):5082-5089.

[28] OSTRYCHARZ E, WASIK U, KEMPINSKA-PODHORODECKA A, et al.Melatonin protects cholangiocytes from oxidative stress-induced proapoptotic and proinflammatory stimuli via miR-132 and miR-34[J].International Journal of Molecular Sciences, 2020, 21(24):9667.
[29] LI Z C, HAN N, LI X, et al.Decreased expression of microRNA-130a correlates with TNF-α in the development of osteoarthritis[J].International Journal of Clinical and Experimental Pathology, 2015, 8(3):2555-2564.

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