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
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摘要
本试验旨在研究脂多糖(LPS)刺激后不同时间断奶仔猪肌肉炎症和肌肉蛋白质降解相关基因表达的变化规律。选择42头(7.1±0.9)kg杜×长×大三元杂断奶仔猪,按注射LPS之前(0 h)和注射LPS后1、2、4、8、12、24 h随机分为7个处理,每个处理6头猪。预试14 d后,腹腔注射100μg/kg体重的LPS。按以上时间点将仔猪屠宰,取背最长肌样品待测。结果表明:背最长肌炎性细胞因子肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、IL-6的mRNA表达量在注射LPS 1～2 h后达到峰值;Toll样受体4信号通路关键基因Toll样受体4(TLR4)、骨髓分化因子88(MyD88),核苷酸结合寡聚域信号通路关键基因核苷酸结合寡聚域2(NOD2)、受体互作蛋白激酶2(RIPK2)的mRNA表达量在注射LPS 2～4 h后达到峰值;肌肉蛋白质降解相关基因叉头转录因子-1(FOXO-1)、FOXO-4、肌肉环指蛋白1(MuRF1)、肌萎缩F-box(MAFbx)的mRNA表达量在注射LPS 12 h后达到峰值。可见,LPS刺激诱导肌肉释放大量炎性细胞因子,使TLR4和NOD炎症信号通路关键基因及肌肉蛋白质降解相关基因mRNA显著表达。
The purpose of this study was to investigate the changes of genes related to muscle inflammation and protein degradation in weaned piglets at different time points after lipopolysaccharide(LPS) challenge. Forty-two weaned piglets(Duroc × Landrace × Yorkshine) with average body weight(7.1±0.9 kg) were randomly divided into seven groups. The piglets were injected with 100 μg/kg BW LPS, and slaughtered at 0 h(before LPS challenge), 1, 2, 4, 8, 12 and 24 h(after LPS challenge). The longissimus dorsi muscle samples were collected for analysis. The results showed that: the mRNA expression of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6 in longissimus dorsi muscle peaked at 1～2 h after LPS injection; The mRNA expression of the key genes in toll-like receptor 4(TLR4) and nucleotide binding oligomerization domin protein(NOD) signaling pathways including TLR4, myeloid differentiation factor 88(MyD88), receptor-interacting protein2(RIPK2) and NOD2 peaked at 2～4 h after LPS injection;(3) The mRNA expression of the key genes related to the muscle protein degradation including Forkhead Box O-1(FOXO-1), FOXO-4, muscle RING finger 1(MuRF1) and muscle atrophy F-box(MAFbx) peaked at 12 h after LPS injection. These results indicate that LPS induces the release of inflammatory cytokines in muscle, and increased mRNA expression of genes related to TLR4 and NOD inflammatory signaling pathways and muscle protein degradation.
The purpose of this study was to investigate the changes of genes related to muscle inflammation and protein degradation in weaned piglets at different time points after lipopolysaccharide(LPS) challenge. Forty-two weaned piglets(Duroc × Landrace × Yorkshine) with average body weight(7.1±0.9 kg) were randomly divided into seven groups. The piglets were injected with 100 μg/kg BW LPS, and slaughtered at 0 h(before LPS challenge), 1, 2, 4, 8, 12 and 24 h(after LPS challenge). The longissimus dorsi muscle samples were collected for analysis. The results showed that: the mRNA expression of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), IL-6 in longissimus dorsi muscle peaked at 1～2 h after LPS injection; The mRNA expression of the key genes in toll-like receptor 4(TLR4) and nucleotide binding oligomerization domin protein(NOD) signaling pathways including TLR4, myeloid differentiation factor 88(MyD88), receptor-interacting protein2(RIPK2) and NOD2 peaked at 2～4 h after LPS injection;(3) The mRNA expression of the key genes related to the muscle protein degradation including Forkhead Box O-1(FOXO-1), FOXO-4, muscle RING finger 1(MuRF1) and muscle atrophy F-box(MAFbx) peaked at 12 h after LPS injection. These results indicate that LPS induces the release of inflammatory cytokines in muscle, and increased mRNA expression of genes related to TLR4 and NOD inflammatory signaling pathways and muscle protein degradation.
引文
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[8]陈少魁,刘玉兰,李权,等.脂多糖刺激对仔猪下丘脑-垂体-肾上腺轴Toll样受体4信号通路关键基因表达的影响[J].动物营养学报,2014,26(11):3356-3361.
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[11]Zhu H,Liu Y,Xie X,et al.Effect of L-arginine on intestinal mucosal immune barrier function in weaned pigs after Escherichia coli LPS challenge[J].Innate Immun,2013,19(3):242-252.
[12]Liu Y,Chen F,Odle J,et al.Fish oil enhances intestinal integrity and inhibits TLR4 and NOD2 signaling pathways in weaned pigs after LPS challenge[J].J Nutr,2012,142(11):2017-2024.
[13]Lee J Y,Zhao L,Hwang D H.Modulation of pattern recognition receptor-mediated inflammation and risk of chronic diseases by dietary fatty acids[J].Nutr Rev,2010,68:38-61.
[14]涂治骁,刘玉兰,吴欢听,等.脂多糖刺激对仔猪肠道、肝脏和肌肉组织NODs信号通路关键基因及其负调控因子mRNA表达的影响[J].中国畜牧杂志,2016,52(11):35-38.
[15]王海波,刘玉兰,李权,等.脂多糖对仔猪下丘脑-垂体-肾上腺轴内应激基因和NOD信号通路关键基因表达的影响[J].中国畜牧杂志,2015,51(1):20-24.
[16]Tamrakar A K,Schertzer J D,Chiu T T,et al.NOD2 activation induces muscle cell-autonomous innate immune responses and insulin resistance[J].Endocrinology,2010,151(12):5624-5637.
[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].武汉:武汉轻工大学,2013.
[3]Takeda K,Akira S.TLR signaling pathways[J].Semin Immunol,2004,16(1):3-9.
[4]Crossland H,Constantin-teodosiu D,Gardiner S M,et al.Apotential role for Akt/FOXO signalling in both protein loss and the impairment of muscle carbohydrate oxidation during sepsis in rodent skeletal muscle[J].J Physiol,2008,586(22):5589-5600.
[5]Wang X,Hu Z,Hu J,et al.Insulin resistance accelerates muscle protein degradation:Activation of the ubiquitin-proteasome pathway by defects in muscle cell signaling[J].Endocrinology,2006,147(9):4160-4168.
[6]Sabroe I,Parker L C,Dower S K,et al.The role of TLR activation in inflammation[J].J Pathology,2008,214(2):126-135.
[7]Fukata M,Vamadevan A S,Abreu M T.Toll-like receptors(TLRs)and Nod-like receptors(NLRs)in inflammatory disorders[J].Semin Immunol,2009,21(4):242-253.
[8]陈少魁,刘玉兰,李权,等.脂多糖刺激对仔猪下丘脑-垂体-肾上腺轴Toll样受体4信号通路关键基因表达的影响[J].动物营养学报,2014,26(11):3356-3361.
[9]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and 2-ΔΔCt method[J].Methods,2001,25(4):402-408.
[10]Mair K H,Sedlak C,K?ser T,et al.The porcine innate immune system:an update[J].Dev Comp Immunol,2014,45(2):321-343.
[11]Zhu H,Liu Y,Xie X,et al.Effect of L-arginine on intestinal mucosal immune barrier function in weaned pigs after Escherichia coli LPS challenge[J].Innate Immun,2013,19(3):242-252.
[12]Liu Y,Chen F,Odle J,et al.Fish oil enhances intestinal integrity and inhibits TLR4 and NOD2 signaling pathways in weaned pigs after LPS challenge[J].J Nutr,2012,142(11):2017-2024.
[13]Lee J Y,Zhao L,Hwang D H.Modulation of pattern recognition receptor-mediated inflammation and risk of chronic diseases by dietary fatty acids[J].Nutr Rev,2010,68:38-61.
[14]涂治骁,刘玉兰,吴欢听,等.脂多糖刺激对仔猪肠道、肝脏和肌肉组织NODs信号通路关键基因及其负调控因子mRNA表达的影响[J].中国畜牧杂志,2016,52(11):35-38.
[15]王海波,刘玉兰,李权,等.脂多糖对仔猪下丘脑-垂体-肾上腺轴内应激基因和NOD信号通路关键基因表达的影响[J].中国畜牧杂志,2015,51(1):20-24.
[16]Tamrakar A K,Schertzer J D,Chiu T T,et al.NOD2 activation induces muscle cell-autonomous innate immune responses and insulin resistance[J].Endocrinology,2010,151(12):5624-5637.