研究论文 RESEARCH PAPER

白术多糖通过线粒体和死亡受体途径缓解脂多糖诱导的雏鹅胸腺细胞凋亡

  • 陈丽君 ,
  • 李冰心 ,
  • 曹楠 ,
  • 付新亮 ,
  • 杨保和 ,
  • 袁明凤 ,
  • 许丹宁 ,
  • 田允波 ,
  • 黄运茂 ,
  • 李婉雁
展开
  • 1. 仲恺农业工程学院动物科技学院, 广州 510225;
    2. 广东省水禽健康养殖重点实验室, 广州 510225;
    3. 云南快大多畜牧科技有限公司, 玉溪 653100
陈丽君(1997—),女,广东湛江人,硕士研究生,动物营养与饲料科学专业。E-mail:2227983785@qq.com

收稿日期: 2022-05-20

  网络出版日期: 2022-12-15

基金资助

国家自然科学基金(32102747);广东省科技发展专项(粤财教[2017]218号);广东省现代农业水禽产业技术体系创新团队(2020KJ137)

Polysaccharide of Atractylodes macrocephala Koidz Relieves Lipopolysaccharide-Induced Apoptosis through Mitochondrial and Death Receptor Pathways in Thymus of Goslings

  • CHEN Lijun ,
  • LI Bingxin ,
  • CAO Nan ,
  • FU Xinliang ,
  • YANG Baohe ,
  • YUAN Mingfeng ,
  • XU Danning ,
  • TIAN Yunbo ,
  • HUANG Yunmao ,
  • LI Wanyan
Expand
  • 1. College of Animal Science & Technology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China;
    2. Guangdong Province Key Laboratory of Waterfowl Healthy Breeding, Guangzhou 510225, China;
    3. Yunnan Kuaidaduo Animal Husbandry Technology Co., Ltd., Yuxi 653100, China

Received date: 2022-05-20

  Online published: 2022-12-15

摘要

本试验旨在通过研究白术多糖(PAMK)对脂多糖(LPS)处理雏鹅胸腺的氧化应激、炎性因子及细胞凋亡水平的影响,探索PAMK是否通过线粒体途径和死亡受体途径对LPS诱导的胸腺细胞凋亡具有缓解作用。试验选用80只1日龄的马岗鹅,随机分为4组,每组5个重复,每个重复4只。对照组(CON组)和脂多糖组(LPS组)饲喂基础饲粮,白术多糖组(PAMK组)和脂多糖+白术多糖组(LPS+PAMK组)在基础饲粮中添加400 mg/kg PAMK;CON组和PAMK组分别在雏鹅第24、26、28日龄腹腔注射1 mL生理盐水;LPS组和LPS+PAMK组分别在雏鹅第24、26、28日龄腹腔注射2 mg/kg LPS溶液。试验期28 d。结果表明:1)苏木精-伊红染色和超微结构观察结果显示,PAMK缓解了LPS引起的胸腺损伤和凋亡。2)与CON组相比,PAMK组的白细胞介素-1β(IL-)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的mRNA相对表达水平无显著差异(P>0.05),白细胞介素-10(IL-10)的mRNA相对表达水平显著下降(P<0.05)。与LPS组相比,LPS+PAMK组的IL-IL-6、TNF-αIL-10的mRNA相对表达水平显著降低(P<0.05)。3)与LPS组相比,PAMK+LPS组过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的mRNA相对表达水平显著升高(P<0.05),谷胱甘肽过氧化物酶(GPX)的mRNA相对表达水平显著降低(P>0.05)。4)与CON组相比,PAMK组B细胞白血病/淋巴瘤-2相关X蛋白(Bax)、肿瘤蛋白63(TP63)的mRNA相对表达水平无显著差异(P>0.05),半胱氨酸天冬氨酸特异性蛋白酶-3(Caspase-3)的mRNA相对表达水平显著降低(P<0.05),B细胞白血病/淋巴瘤-2(Bcl-2)、半胱氨酸天冬氨酸特异性蛋白酶-8(Caspase-8)的mRNA相对表达水平显著升高(P<0.05)。与LPS组相比,LPS+PAMK组BaxCaspase-3的mRNA相对表达水平显著降低(P<0.05),Bcl-2、Caspase-8、TP63的mRNA相对表达水平表达显著升高(P<0.05)。5)与CON组相比,PAMK组Bax、Caspase-8的蛋白相对表达水平无显著差异(P>0.05),Bcl-2的蛋白相对表达水平显著下降(P<0.05)。与LPS组相比,LPS+PAMK组Bax、Caspase-8的蛋白相对表达水平显著下降(P<0.05),Bcl-2的蛋白相对表达水平显著升高(P<0.05)。由此可见,PAMK可抑制LPS引起的氧化应激、炎性因子水平上升,通过线粒体及死亡受体途径缓解雏鹅胸腺细胞凋亡,并维持胸腺组织的结构完整性。

本文引用格式

陈丽君 , 李冰心 , 曹楠 , 付新亮 , 杨保和 , 袁明凤 , 许丹宁 , 田允波 , 黄运茂 , 李婉雁 . 白术多糖通过线粒体和死亡受体途径缓解脂多糖诱导的雏鹅胸腺细胞凋亡[J]. 动物营养学报, 2022 , 34(12) : 8049 -8060 . DOI: 10.3969/j.issn.1006-267x.2022.12.052

Abstract

This experiment was conducted to study the effects of polysaccharide of Atractylodes macrocephala Koidz (PAMK) on oxidative stress, inflammatory factors and apoptosis level in thymus of goslings treated with lipopolysaccharide (LPS), to find that whether PAMK relieved LPS-induced apoptosis through the mitochondrial and death receptor pathways in thymus of goslings. A total of 80 one-day-old healthy goslings were selected and randomly divided into 4 groups with 5 replicates per group and 4 goslings per replicate. Goslings in control group (CON group) and lipopolysaccharide group (LPS group) were fed the basal diets, and others in polysaccharide of Atractylodes macrocephala Koidz group (PAMK group) and lipopolysaccharide+ polysaccharide of Atractylodes macrocephala Koidz group (LPS+PAMK group) were fed basal diets supplemented with 400 mg/kg PAMK. Goslings in CON group and PAMK group were intraperitoneal injected 1 mL normal saline on days 24, 26 and 28, respectively, and goslings in LPS group and LPS+PAMK group were intraperitoneal injected 2 mg/kg LPS solution on days 24, 26 and 28, respectively. The experiment lasted for 28 days. The results showed as follows:1) the hematoxylin-eosin staining and ultrastructural observation showed that PAMK relieved the LPS-induced injury and apoptosis in thymus. 2) Compared with the CON group, the mRNA relative expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) of PAMK group were no significant difference (P>0.05), and the interleukin-10 (IL-10) mRNA relative expression level was significantly decreased (P<0.05). Compared with the LPS group, the mRNA relative expression levels of IL-1β, IL-6, TNF-α and IL-10 of LPS+PAMK group were significantly decreased (P<0.05). 3) Compared with the LPS group, the mRNA relative expression levels of catalase (CAT), superoxide dismutase (SOD) of LPS+PAMK group were significantly increased (P<0.05), and the glutathione peroxidase (GPX) mRNA relative expression level was significantly decreased (P<0.05). 4) Compared with the CON group, the mRNA relative expression levels of B-cell lymphoma/leukaemia-2-associated X protein (Bax) and tumor protein 63 (TP63) of PAMK group were no significant difference (P>0.05), the cysteinyl aspartate specific proteinase-3 (Caspase-3) mRNA relative expression level was significantly decreased (P<0.05), and the mRNA relative expression levels of B-cell lymphoma/leukaemia-2 (Bcl-2) and cysteinyl aspartate specific proteinase-8 (Caspase-8) were significantly increased (P<0.05). Compared with the LPS group, the mRNA relative expression levels of Bax and Caspase-3 of LPS+PAMK group were significantly decreased (P<0.05), and the mRNA relative expression levels of Bcl-2, Caspase-8 and TP63 were significantly increased (P<0.05). 5) Compared with the CON group, the protein relative expression levels of Bax and Caspase-8 of PAMK group were no significant difference (P>0.05), and the Bcl-2 protein relative expression level was significantly decreased (P<0.05). Compared with the LPS group, the protein relative expression levels of Bax and Caspase-8 of LPS+PAMK group were significantly decreased (P<0.05), and the Bcl-2 protein relative expression level was significantly increased (P<0.05). In conclusion, the PAMK can inhibit the increase of oxidative stress and inflammatory factor levels induced by LPS, alleviate the thymus apoptosis of goslings through the mitochondrial and death receptor pathways, and maintain the structural integrity of thymus tissue.

参考文献

[1] SAVINO W.The thymus is a common target organ in infectious diseases[J].PLoS Pathogens, 2006, 2(6):e62.
[2] RAVIOLA E, KARNOVSKY M J.Evidence for a blood-thymus barrier using electron-opaque tracers[J].Journal of Experimental Medicine, 1972, 136(3):466-498.  
[3] ZHAO H J, HE Y, LI S W, et al.Subchronic arsenism-induced oxidative stress and inflammation contribute to apoptosis through mitochondrial and death receptor dependent pathways in chicken immune organs[J].Oncotarget, 2017, 8(25):40327-40344.  
[4] FAN G W, JIANG X R, WU X Y, et al.Anti-inflammatory activity of tanshinone ⅡA in LPS-stimulated RAW264.7 macrophages via miRNAs and TLR4-NF-κB pathway[J].Inflammation, 2016, 39(1):375-384.  
[5] NIGHOT M, AL-SADI R, GUO S H, et al.Lipopolysaccharide-induced increase in intestinal epithelial tight permeability is mediated by Toll-like receptor 4/myeloid differentiation primary response 88 (MyD88) activation of myosin light chain kinase expression[J].The American Journal of Pathology, 2017, 187(12):2698-2710.  
[6] QIN X Y, JIANG X R, JIANG X, et al.Micheliolide inhibits LPS-induced inflammatory response and protects mice from LPS challenge[J].Scientific Reports, 2016, 6(1):23240.
[7] DENG X M, LUYENDYK J P, GANEY P E, et al.Inflammatory stress and idiosyncratic hepatotoxicity:hints from animal models[J].Pharmacological Reviews, 2009, 61(3):262-282.  
[8] SEO H Y, KIM M K, LEE S H, et al.Kahweol ameliorates the liver inflammation through the inhibition of NF-κB and STAT3 activation in primary Kupffer cells and primary hepatocytes[J].Nutrients, 2018, 10(7):863.
[9] 王华.细菌脂多糖诱导小鼠急性凋亡性肝损伤的分子机制[D].硕士学位论文.合肥:安徽医科大学, 2009. WANG H.Molecular mechanisms of lipopolysaccharide-induced acute apoptotic liver injury in mice[D].Master's Thesis.Hefei:Anhui Medical University, 2009.(in Chinese)
[10] ZHANG C, WANG F, ZHANG Y X, et al.Celecoxib prevents pressure overload-induced cardiac hypertrophy and dysfunction by inhibiting inflammation, apoptosis and oxidative stress[J].Journal of Cellular and Molecular Medicine, 2016, 20(1):116-127.  
[11] GALLUZZI L, KEPP O, TROJEL-HANSEN C, et al.Mitochondrial control of cellular life, stress, and death[J].Circulation Research, 2012, 111(9):1198-1207.  
[12] ELMORE S.Apoptosis:a review of programmed cell death[J].Toxicologic Pathology, 2007, 35(4):495-516.  
[13] GALLUZZI L, VITALE I, ABRAMS J M, et al.Molecular definitions of cell death subroutines:recommendations of the Nomenclature Committee on Cell Death 2012[J].Cell Death and Differentiation, 2012, 19(1):107-120.  
[14] ELROD H A, SUN S Y.Modulation of death receptors by cancer therapeutic agents[J].Cancer Biology & Therapy, 2008, 7(2):163-173.  
[15] XU D N, LI W Y, HUANG Y M, et al.The effect of selenium and polysaccharide of Atractylodes macrocephala Koidz. (PAMK) on immune response in chicken spleen under heat stress[J].Biological Trace Element Research, 2014, 160(2):232-237.  
[16] FAN W T, ZHANG S J, HAO P, et al.Structure characterization of three polysaccharides and a comparative study of their immunomodulatory activities on chicken macrophage[J].Carbohydrate Polymers, 2016, 153:631-640.
[17] WANG X T, LI L H, RAN X K, et al.What caused the changes in the usage of Atractylodis macrocephalae Rhizoma from ancient to current times?[J].Journal of Natural Medicines, 2016, 70(1):36-44.  
[18] LI W Y, XIANG X L, LI B X, et al.PAMK relieves LPS-induced enteritis and improves intestinal flora disorder in goslings[J].Evidence-Based Complementary and Alternative Medicine, 2021, 2021:9721353.
[19] LI W Y, GUO S X, XU D N, et al.Polysaccharide of Atractylodes macrocephala Koidz (PAMK) relieves immunosuppression in cyclophosphamide-treated geese by maintaining a humoral and cellular immune balance[J].Molecules, 2018, 23(4):932.
[20] XU D N, LI B X, CAO N, et al.The protective effects of polysaccharide of Atractylodes macrocephala Koidz (PAMK) on the chicken spleen under heat stress via antagonizing apoptosis and restoring the immune function[J].Oncotarget, 2017, 8(41):70394-70405.  
[21] TANAKA T, NARAZAKI M, KISHIMOTO T.IL-6 in inflammation, immunity, and disease[J].Cold Spring Harbor Perspectives in Biology, 2014, 6(10):a016295.
[22] WHITE E.Life, death, and the pursuit of apoptosis[J].Genes & Development, 1996, 10(1):1-15.  
[23] ROULSTON A, MARCELLUS R C, BRANTON P E.Viruses and apoptosis[J].Annual Review of Microbiology, 1999, 53:577-628.
[24] 付晶, 赵丹, 林桐, 等.白术多糖对环磷酰胺致雏鸡肝脏细胞凋亡中线粒体通路的影响[J].东北农业大学学报, 2019, 50(4):63-70. FU J, ZHAO D, LIN T, et al.Effect of PAMK on apoptosis through mitochondrial pathway in chicken liver induced by CTX[J].Journal of Northeast Agricultural University, 2019, 50(4):63-70.(in Chinese)
[25] LINKERMANN A, STOCKWELL B R, KRAUTWALD S, et al.Regulated cell death and inflammation:an auto-amplification loop causes organ failure[J].Nature Reviews Immunology, 2014, 14(11):759-767.  
[26] KARIN M, LIN A N.NF-κB at the crossroads of life and death[J].Nature Immunology, 2002, 3(3):221-227.  
[27] LI L, WAN G W, HAN B, et al.Echinacoside alleviated LPS-induced cell apoptosis and inflammation in rat intestine epithelial cells by inhibiting the mTOR/STAT3 pathway[J].Biomedicine & Pharmacotherapy, 2018, 104:622-628.
[28] LIANG Z Y, REN C F.Retracted:emodin attenuates apoptosis and inflammation induced by LPS through up-regulating lncRNA TUG1 in murine chondrogenic ATDC5 cells[J].Biomedicine & Pharmacotherapy, 2018, 103:897-902.
[29] MA H L, SU L, HE X Y, et al.Loss of HMBOX1 promotes LPS-induced apoptosis and inhibits LPS-induced autophagy of vascular endothelial cells in mouse[J].Apoptosis, 2019, 24(11/12):946-957.
[30] SONG D Q, LIU J, WANG F, et al.Procyanidin B2 inhibits lipopolysaccharide-induced apoptosis by suppressing the Bcl-2/Bax and NF-κB signalling pathways in human umbilical vein endothelial cells[J].Molecular Medicine Reports, 2021, 23(4):267.
[31] LI J C, QIAO Z J, HU W Y, et al.Baicalin mitigated Mycoplasma gallisepticum-induced structural damage and attenuated oxidative stress and apoptosis in chicken thymus through the Nrf2/HO-1 defence pathway[J].Veterinary Research, 2019, 50(1):83.
[32] HIRSOVA P, GORES G J.Death receptor-mediated cell death and proinflammatory signaling in nonalcoholic steatohepatitis[J].Cellular and Molecular Gastroenterology and Hepatology, 2015, 1(1):17-27.  
[33] GRESSNER O, SCHILLING T, LORENZ K, et al.TAp63α induces apoptosis by activating signaling via death receptors and mitochondria[J].The EMBO Journal, 2005, 24(13):2458-2471.  
文章导航

/