Polysaccharide of Atractylodes macrocephala Koidz Regulates Lymphocyte Immune Function in Chick Spleen by Toll-Like Receptor 4/ Nuclear Factor-Kappa B Signal Pathway

  • LI Wanyan ,
  • CAO Nan ,
  • TIAN Yunbo ,
  • XIANG Xuelian ,
  • LI Bingxin ,
  • HUANG Yunmao ,
  • XU Danning
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  • 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

Received date: 2019-04-15

  Online published: 2019-11-19

Abstract

The aim of this study was to investigate the effects of polysaccharide of Atractylodes macrocephala Koidz (PAMK) on lymphocyte immune function in chick spleen and possible regulatory mechanisms. The spleen lymphocytes of 21-day-old chicks were isolated and cultured in vitro, the lymphocytes were transfected by siRNA primers and the expression of Toll-like receptor 4 (TLR4) was interfered. On that basis, PAMK with the concentration of 12.5 mg/L was used to treat transfected and non-transfected lymphocytes. After 36 h, the mRNA expression levels of TLR4 and its signaling pathway downstream genes[such as myeloid differentiation factor 88 (MyD88), nuclear factor-kappa B (NF-κB) inhibitor kinase (IKK), NF-κB inhibitory protein (IκB)] and cytokines[such as interleukin-2 (IL-2), interleukin-4 (IL-4), interferon-γ (IFN-γ), lipopolysaccharide induced tumor necrosis factor-α (LITAF)] were detected. In addition, the content of NF-κB in lymphocyte nuclear protein was detected by electrophoretic mobility shift assay (EMSA) method. The results showed that PAMK could significantly increase the mRNA expression levels of TLR4 and its signaling pathway downstream genes IKK, IκB, cytokines IL-2 and LITAF in non-transfected lymphocytes (P<0.05), and significantly increase the mRNA expression levels of TLR4 in transfected lymphocytes (P<0.05). In addition, PAMK could also promote the expression of cytokines[IL-2 (P>0.05), IFN-γ (P<0.05), LITAF (P>0.05)] secreted by type 1 helper T cells (Th1) in transfected lymphocytes to different degrees, there was no significant promoting effect on the expression of cytokine (IL-4) secreted by type 2 helper T cells (Th2) (P>0.05). EMSA results showed that PAMK could promote NF-κB into the nuclear in transfected and non-transfected lymphocytes, and significantly increase the content of NF-κB in nuclear protein (P<0.05). In summary, PAMK activates TLR4/NF-κB signaling pathway to promote the expression of related genes, and finally promotes NF-κB into the nuclear to regulate the transcription level of cytokines, and then regulates lymphocyte immune function in chick spleen.

Cite this article

LI Wanyan , CAO Nan , TIAN Yunbo , XIANG Xuelian , LI Bingxin , HUANG Yunmao , XU Danning . Polysaccharide of Atractylodes macrocephala Koidz Regulates Lymphocyte Immune Function in Chick Spleen by Toll-Like Receptor 4/ Nuclear Factor-Kappa B Signal Pathway[J]. Chinese Journal of Animal Nutrition, 2019 , 31(11) : 5192 -5201 . DOI: 10.3969/j.issn.1006-267x.2019.11.035

References

[1] 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.
[2] LI X J,LIU F,LI Z,et al.Atractylodes macrocephala polysaccharides induces mitochondrial-mediated apoptosis in glioma C6 cells[J].International Journal of Biological Macromolecules,2014,66:108-112.
[3] SHU Y T,KAO K T,WENG C S.In vitro antibacterial and cytotoxic activities of plasma-modified polyethylene terephthalate nonwoven dressing with aqueous extract of rhizome Atractylodes macrocephala[J].Materials Science and Engineering:C,2017,77:606-612.
[4] WANG J C,CHEN J,PAN K W.Effect of exogenous abscisic acid on the level of antioxidants in Atractylodes macrocephala Koidz under lead stress[J].Environmental Science and Pollution Research,2013,20(3):1441-1449.  
[5] XU D N,TIAN Y B.Selenium and polysaccharides of Atractylodes macrocephala Koidz play different roles in improving the immune response induced by heat stress in chickens[J].Biological Trace Element Research,2015,168(1):235-241.  
[6] GUO L,SUN Y L,WANG A H,et al.Effect of polysaccharides extract of Rhizoma Atractylodis Macrocephalae on thymus,spleen and cardiac indexes,caspase-3 activity ratio,Smac/DIABLO and HtrA2/Omi protein and mRNA expression levels in aged rats[J].Molecular Biology Reports,2012,39(10):9285-9290.  
[7] 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):932-947.  
[8] LIU J,CHEN X,YUE C J,et al.Effect of selenylation modification on immune-enhancing activity of Atractylodes macrocephala polysaccharide[J].International Journal of Biological Macromolecules,2015,72:1435-1440.
[9] SUN W J,MENG K,QI C H,et al.Immune-enhancing activity of polysaccharides isolated from Atractylodis macrocephalae Koidz[J].Carbohydrate Polymers,2015,126:91-96.
[10] SON Y O,KOOK S H,LEE J C.Glycoproteins and polysaccharides are the main class of active constituents required for lymphocyte stimulation and antigen-specific immune response induction by traditional medicinal herbal plants[J].Journal of Medicinal Food,2017,20(10):1011-1021.  
[11] ZHOU L J,LIU Z J,WANG Z X,et al.Astragalus polysaccharides exerts immunomodulatory effects via TLR4-mediated MyD88-dependent signaling pathway in vitro and in vivo[J].Science Report,2017,7:44822.
[12] 刘瑞.黄芪多糖影响雏鸡免疫器官TLR4信号转导通路的研究[D].硕士学位论文.哈尔滨:东北农业大学,2015.
[13] ZHAO T,FENG Y,LI J,et al.Schisandra polysaccharide evokes immunomodulatory activity through TLR 4-mediated activation of macrophages[J].International Journal of Biological Macromolecules,2014,65:33-40.
[14] LIU L,YUAN S S,LONG Y,et al.Immunomodulation of Rheum tanguticum polysaccharide (RTP) on the immunosuppressive effects of dexamethasone (DEX) on the treatment of colitis in rats induced by 2,4,6-trinitrobenzene sulfonic acid[J].International Immunopharmacology,2009,9(13/14):1568-1577.
[15] GONG A G,ZHANG L M,LAM C T,et al.Polysaccharide of Danggui Buxue Tang,an ancient Chinese herbal decoction,induces expression of pro-inflammatory cytokines possibly via activation of NFκB signaling in cultured RAW 264.7 cells[J].Phytotherapy Research,2017,31(2):274-283.  
[16] JIN B,SUN T,YU X H,et al.The effects of TLR activation on T-cell development and differentiation[J].Clinical and Developmental Immunology,2012,2012:836485.
[17] PÓCIENNIKOWSKA A,HROMADA-JUDYCKA A,BORZECKA K,et al.Co-operation of TLR4 and raft proteins in LPS-induced pro-inflammatory signaling[J].Cellular and Molecular Life Science,2015,72(3):557-581.  
[18] 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.  
[19] HUANG X Q,XIE Z J,LIU F F,et al.Dihydroartemisinin inhibits activation of the Toll-like receptor 4 signaling pathway and production of type Ⅰ interferon in spleen cells from lupus-prone MRL/lpr mice[J].International Immunopharmacology,2014,22(1):266-272.  
[20] JI G Q,CHEN R Q,ZHENG J X.Macrophage activation by polysaccharides from Atractylodes macrocephala Koidz through the nuclear factor-κB pathway[J].Pharmaceutical Biology,2015,53(4):512-517.  
[21] PI J,WANG Y Y,ZHU H Y,et al.Immunomodulatory effects of polysaccharide compounds in macrophages revealed by high resolution AFM[J].Scanning,2016,38(6):792-801.  
[22] TANAKA K,INA A.Structure elucidation of acylsucrose derivatives from Atractylodes lanceae rhizome and Atractylodes rhizome[J].Natural Product Communication,2009,4(8):1095-1098.
[23] 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.  
[24] 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.  
[25] 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.
[26] CEN X H,LIU S W,CHENG K.The role of Toll-like receptor in inflammation and tumor immunity[J].Frontiers in Pharmacology,2018,9:878.
[27] AHMED-HASSAN H,ABDUL-CADER M,SABRY M,et al.Toll-like receptor (TLR) 4 signalling induces myeloid differentiation primary response gene (MYD) 88 independent pathway in avian species leading to type Ⅰ interferon production and antiviral response[J].Virus Research,2018,256:107-116.
[28] HACKAM D J,SODHI C P.Toll-like receptor-mediated intestinal inflammatory imbalance in the pathogenesis of necrotizing enterocolitis[J].Cellular and Molecular Gastroenterology and Hepatology,2018,6(2):229-238.  
[29] KARNATI H K,PASUPULETI S R,KANDI R,et al.TLR-4 signalling pathway:MyD88 independent pathway up-regulation in chicken breeds upon LPS treatment[J].Veterinary Research Communications,2015,39(1):73-78.  
[30] ZHOU L J,LIU Z J,WANG Z X,et al.Astragalus polysaccharides exerts immunomodulatory effects via TLR4-mediated MyD88-dependent signaling pathway in vitro and in vivo[J].Science Report,2017,7:44822.
[31] HAN S B,YOON Y D,AHN H J,et al.Toll-like receptor-mediated activation of B cells and macrophages by polysaccharide isolated from cell culture of Acanthopanax senticosus[J].International Immunopharmacology,2003,3(9):1301-1312.  
[32] MENG J J,HU X F,SHAN F P,et al.Analysis of maturation of murine dendritic cells (DCs) induced by purified Ganoderma lucidum polysaccharides (GLPs)[J].International Journal of Biological Macromolecules,2011,49(4):693-699.  
[33] WEINLICH R,BORTOLUCI K R,CHEHAB C F,et al.TLR4/MYD88-dependent,LPS-induced synthesis of PGE2 by macrophages or dendritic cells prevents anti-CD3-mediated CD95L upregulation in T cells[J].Cell Death and Differentiation,2008,15(12):1901-1909.  
[34] MA L,ZHANG L W,ZHUANG Y,et al.Serum levels of IL-33 and correlation with IL-4,IL-17A,and hypergammaglobulinemia in patients with autoimmune hepatitis[J].Mediators of Inflammation,2018,2018:7964654.
[35] TRENTIN-SONODA M,DA SILVA R,KMIT F,et al.Knockout of Toll-like receptors 2 and 4 prevents renal ischemia-reperfusion-induced cardiac hypertrophy in mice[J].PLoS One,2015,10(10):e139350.
[36] HOU Y C,WU J M,WANG M Y,et al.Modulatory effects of Astragalus polysaccharides on T-cell polarization in mice with polymicrobial sepsis[J].Mediators of Inflammation,2015,2015:826319.
[37] LIU H,ZHANG W,DONG S C,et al.Protective effects of sea buckthorn polysaccharide extracts against LPS/d-GalN-induced acute liver failure in mice via suppressing TLR4-NF-κB signaling[J].Journal of Ethnopharmacology,2015,176:69-78.
[38] HAN J,LIU L X,YU N,et al.Polysaccharides from Acanthopanax senticosus enhances intestinal integrity through inhibiting TLR4/NF-κB signaling pathways in lipopolysaccharide-challenged mice[J].Animal Science Journal,2016,87(8):1011-1018.  
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