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香菇多糖对脂多糖刺激的仔猪空肠细胞死亡信号通路相关基因表达的影响

  • 梁天增 ,
  • 汪龙梅 ,
  • 余程 ,
  • 王秀英 ,
  • 刘玉兰 ,
  • 朱惠玲
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  • 武汉轻工大学动物营养与饲料科学湖北重点实验室, 武汉 430023
梁天增(1995-),男,河南平顶山人,硕士研究生,从事猪的营养与免疫的研究。E-mail:2523107520@qq.com

收稿日期: 2019-08-24

  网络出版日期: 2020-02-21

基金资助

武汉市应用基础前沿项目(2018020401011304)

Effects of Lentinan on Jejunal Cell Death Signaling Pathway Related Gene Expression of Piglets Challenged with Lipopolysaccharide

  • LIANG Tianzeng ,
  • WANG Longmei ,
  • YU Cheng ,
  • WANG Xiuying ,
  • LIU Yulan ,
  • ZHU Huiling
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  • Hubei Key Laboratory of Animal Nutrition and Feed Science, Wuhan Polytechnic University, Wuhan 430023, China

Received date: 2019-08-24

  Online published: 2020-02-21

摘要

本试验旨在探讨香菇多糖对脂多糖(LPS)刺激的仔猪空肠细胞死亡信号通路相关基因表达的影响。选取24头健康的28日龄"杜×长×大"断奶仔猪,平均体重为(8.12±0.19)kg,随机分为4组,每组6个重复,每个重复1头猪。试验采用2×2因子设计,主因子分别为饲粮处理(基础饲粮或基础饲粮中添加0.02%的香菇多糖)和免疫应激处理(注射生理盐水或LPS)。试验第28天,免疫应激组仔猪腹膜注射100 μg/kg体重(BW)的LPS或生理盐水。注射LPS或生理盐水4 h后,仔猪麻醉屠宰,取空肠样品待测。试验期为28 d。结果表明:1)LPS刺激导致仔猪空肠绒毛萎缩,肠上皮脱落,肠道形态受损;饲粮添加香菇多糖缓解了LPS刺激所导致的肠道形态结构损伤。2)LPS刺激显著提高了空肠程序性坏死信号通路关键基因受体互作蛋白激酶3(RIP3)mRNA的相对表达量(P<0.05),显著降低了动力相关蛋白1(Drp1)和混合系列蛋白激酶结构域样蛋白(MLKL)mRNA的相对表达量(P<0.05)。饲粮添加香菇多糖显著降低了空肠程序性坏死信号通路关键基因受体互作蛋白激酶1(RIP1)、Fas死亡结构域相关蛋白(FADD)mRNA的相对表达量(P<0.05)。3)LPS刺激显著降低了空肠焦亡信号通路关键基因凋亡相关斑点样蛋白(ASC)、半胱天冬酶(Caspase)-1、白细胞介素-18(IL-18)以及凋亡信号通路关键基因Caspase-9、B淋巴细胞瘤-2相关X蛋白(Bax)mRNA的相对表达量(P<0.05),显著提高了空肠焦亡信号通路关键基因核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、消皮素D(GSDMD)及凋亡信号通路关键基因Caspase-8 mRNA的相对表达量(P<0.05)。饲粮添加香菇多糖显著降低了空肠焦亡信号通路关键基因NLRP3 mRNA的相对表达量(P<0.05)及凋亡信号通路关键基因Caspase-9和原癌基因蛋白xl(Bcl-xl)mRNA的相对表达量(P<0.05)。4)LPS刺激显著降低了空肠自噬信号通路关键基因自噬相关基因12(Atg12)、微管相关蛋白1轻链3(LC3)、哺乳动物雷帕霉素靶蛋白(mTOR)和Unc-51样激酶1(ULK1)mRNA的相对表达量(P<0.05)。饲粮添加香菇多糖显著降低了空肠自噬信号通路关键基因Atg12、LC3和ULK1 mRNA的相对表达量(P<0.05)。由此可见,饲粮添加香菇多糖是通过调控程序性坏死、焦亡和自噬信号通路关键基因,从而维持肠道完整性。

本文引用格式

梁天增 , 汪龙梅 , 余程 , 王秀英 , 刘玉兰 , 朱惠玲 . 香菇多糖对脂多糖刺激的仔猪空肠细胞死亡信号通路相关基因表达的影响[J]. 动物营养学报, 2020 , 32(2) : 915 -924 . DOI: 10.3969/j.issn.1006-267x.2020.02.047

Abstract

The purpose of this experiment was to investigate the effects of lentinan on jejunal cell death signaling pathway related gene expression of piglets challenged with lipopolysaccharide (LPS). Twenty-four healthy 28-day-old "Duroc×Landrace×Yorkshine" weaned piglets with average body weight of (8.12±0.19) kg were randomly divided into 4 groups with 6 replicates per group and 1 pig per replicate. The experiment consisted of a 2×2 factorial design, and the main factors included dietary treatment (basic diet or basic diet supplemented with 0.02% lentinan) and immunological stress treatment (injection of saline or LPS). On days 28 of the experiment, piglets in immune groups were injected with 100 μg/kg BW of LPS or saline. After 4 h LPS or saline administration, the animals were slaughtered and the jejunal samples were collected for analysis. The experiment lasted for 28 days. The results showed as follows:1) LPS challenge caused jejunal villus atrophy, intestinal epithelial exfoliation and intestinal morphology damage of pigltes; dietary lentinan supplementation alleviated intestinal morphology injury induced by LPS. 2) LPS challenge significantly increased the mRNA relative expression of necroptotic signal pathway key gene of receptor interacting protein kinase 3 (RIP3) in jejunum (P<0.05), and significantly decreased the mRNA relative expressions of dynamic related protein 1 (Drp1) and mixed lineage kinase domain-like (MLKL) (P<0.05). Dietary lentinan supplementation significantly decreased the mRNA relative expressions of necroptotic signaling pathway key genes of receptor interacting protein kinase 1 (RIP1) and Fas associated protein with death domain (FADD) in jejunum (P<0.05). 3) LPS challenge significantly decreased the mRNA relative expressions of pyroptosis signaling pathway key genes of apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate specific proteinase (Caspase)-1, interleukin-18 (IL-18) and apoptosis signaling pathway key genes of Caspase-9, B cell lymphoma/leukemia-2 associated X protein (Bax) in jejunum (P<0.05), and significantly increased the mRNA relative expressions of pyroptosis signaling pathway key genes of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), gasdermin D (GSDMD) and apoptosis signaling pathway key genes of Caspase-8 (P<0.05). Dietary lentinan supplementation significantly decreased the mRNA relative expressions of pyroptosis signaling pathway key gene of NLRP3 and apoptosis signaling pathway key genes of Caspase-9, B-cell lymphoma-xl (Bcl-xl) in jejunum (P<0.05). 4) LPS challenge significantly decreased the mRNA relative expressions of autophagic signaling pathway key genes of autophagy related gene 12 (Atg12), microtubule-associated protein 1 light chain 3 (LC3), mammalian target of rapamycin (mTOR) and UNC-51 like kinase 1 (ULK1) in jejunum (P<0.05). Dietary lentinan supplementation significantly decreased the mRNA relative expressions of autophagy signaling pathway key genes of Atg12, LC3 and ULK1 in jejunum (P<0.05). In conclusion, dietary lentinan supplementation maintain intestinal integrity by regulating key genes in necroptosis, pyroptosis and autophagy signaling pathways.

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