白术多糖可能通过Toll样受体4信号通路缓解环磷酰胺诱导的雏鹅肝脏损伤
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摘要
本试验旨在探讨白术多糖(PAMK)对环磷酰胺(CTX)诱导的雏鹅肝脏损伤的影响及可能的作用机制。试验选用健康的1日龄马岗鹅雏鹅180只,采用单因素完全随机分组试验设计,随机分成3组(对照组、CTX组、PAMK+CTX组),每组3个重复,每个重复20只。PAMK+CTX组从4日龄开始在基础饲粮中添加400 mg/kg的PAM K,直至试验结束。19、20、21日龄,对照组雏鹅腿肌注射0.5 mL生理盐水,CTX组、PAMK+CTX组雏鹅分别腿肌注射40 mg/kg BW的CTX溶液。试验期28 d。结果表明:1)与对照组相比,CTX组肝脏指数显著升高(P<0.05),胆囊指数显著下降(P<0.05);PAMK+CTX组肝脏指数和胆囊指数与对照组相比无显著差异(P>0.05)。2) CTX组和PAMK+CTX组肝脏中的8-羟基脱氧鸟苷含量显著高于对照组(P<0.05)。3) CTX可导致肝脏损伤,出现组织纤维化、炎性小灶增多及胆汁淤积等病变,肝细胞DNA氧化损伤;而PAMK可减轻肝脏组织受损程度,减少坏死肝细胞数量。4)与对照组相比,CTX组肝脏中丙二醛(MDA)含量显著升高(P<0.05),肝脏中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、诱导型一氧化氮合酶(i NOS)活性显著降低(P<0.05)。PAMK+CTX组肝脏中MDA含量和GSH-Px、i NOS活性与对照组无显著差异(P>0.05)。5) CTX组肝脏Toll样受体4(TLR4)、肿瘤坏死因子受体相关因子6(TRAF6)、细胞外信号调节激酶激酶1(MEKK1)、丝裂原活化蛋白激酶激酶3(MKK3)、p38丝裂原活化蛋白激酶(p38MAPK)、核因子κB抑制蛋白α(IκBα)、IκB激酶α(IKKα) mRNA的相对表达量显著低于对照组(P<0.05)。PAMK+CTX组肝脏TLR4、髓样分化因子88(MyD88)、MEKK1、MKK3、丝裂原活化蛋白激酶激酶6(MKK6)、IκB激酶ε(IKKε)、IκBαmRNA的相对表达量显著高于CTX组(P<0.05),肝脏MyD88、MKK6 mRNA的相对表达量显著高于对照组(P<0.05),肝脏TRAF6、MEKK1、MKK3、p38MAPK mRNA的相对表达量显著低于对照组(P>0.05)。由此可见,PAM K对CTX诱导的雏鹅肝损伤的调节作用可能是通过TLR4信号通路来实现的。
This trial was conducted to evaluate the effects and possible mechanism of polysaccharide of Atractylodes macrocephala Koidz(PAMK) on liver injury induced by cyclophosphamide(CTX) in goslings.A total of 180 healthy one-day old Magang goslings were randomly divided into 3 groups(control group,CTX group and PAMK+CTX group) with 3 replicates per group and 20 geese per replicate in a single-factor completely randomized experimental design.Goslings in PAMK+CTX group were fed the basal diet supplemented with 400 mg/kg PAMK from 4 days of age to the end of the trial.On 19,20 and 21 days of age,goslings in the control group were intramuscular injected 0.5 mL normal saline,and goslings in CTX group and PAMK+CTX group were intramuscular injected 40 mg/kg BW CTX solution,respectively.The trail lasted for 28 days.The results showed as follows:1) compared with the control group,the liver index of control group was significantly increased(P<0.05),and the gallbladder index was significantly decreased(P<0.05).The liver index and gallbladder index of PAMK+CTX group had no significant difference compared with the control group(P>0.05).2) The liver 8-hydroxy-deoxyguanosine content of CTX group and PAMK+CTX group was significantly higher than that of the control group(P<0.05).3) The CTX could cause liver pathological damages,DNA oxidative damage of hepatic cells and some lesions in the liver,such as tissue fibrosis,inflammatory small focus,cholestasis et al.The PAMK could alleviate extent of damage of liver tissue,reduce the number of necrotic hepatocytes.4) Compared with the control group,the liver malondialdehyde(MDA) content of CTX group was significantly increased(P<0.05),and the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT),inducible nitric oxide synthase(iNOS) in liver were significantly decreased(P<0.05).The MDA content and GSH-Px,iNOS activities in liver of PAMK+CTX group had no significant difference compared with the control group(P>0.05).5) The mRNA relative expressions of Tolllike receptor 4(TLR4),tumor necrosis factor receptor-associated factor 6(TRAF6),mitogen activated protein kinase kinase 1(MEKK1),mitogen-activated protein kinase kinase 3(MKK3),p38 mitogen-activated protein kinase(p38 MAPK),inhibitor of nuclear factor-κB α(IκBα) and inhibitor of nuclear factor kappa-B kinase α(IKKα) in liver of CTX group were significantly lower than those of the control group(P<0.05).The mRNA relative expressions of TLR4,myeloid differentiation primary response(MyD88),MEKK1,MKK3,mitogen-activated protein kinase kinase 6(MKK6),inhibitor of nuclear factor kappa-B kinase ε(IKKε) and IκBαin liver of PAMK + CTX group were significantly higher than those of CTX group(P<0.05),the mRNA relative expressions of MyD88 and MKK6 in liver were significantly higher than those of the control group(P<0.05),and the mRNA relative expressions of TRAF6,MEKK1,MKK3 and p38 MAPK in liver were significantly lower than those of the control group(P<0.05).These results suggested that PAMK may regulate liver injury induced by CTX in goslings via TLR4 signaling pathway.
This trial was conducted to evaluate the effects and possible mechanism of polysaccharide of Atractylodes macrocephala Koidz(PAMK) on liver injury induced by cyclophosphamide(CTX) in goslings.A total of 180 healthy one-day old Magang goslings were randomly divided into 3 groups(control group,CTX group and PAMK+CTX group) with 3 replicates per group and 20 geese per replicate in a single-factor completely randomized experimental design.Goslings in PAMK+CTX group were fed the basal diet supplemented with 400 mg/kg PAMK from 4 days of age to the end of the trial.On 19,20 and 21 days of age,goslings in the control group were intramuscular injected 0.5 mL normal saline,and goslings in CTX group and PAMK+CTX group were intramuscular injected 40 mg/kg BW CTX solution,respectively.The trail lasted for 28 days.The results showed as follows:1) compared with the control group,the liver index of control group was significantly increased(P<0.05),and the gallbladder index was significantly decreased(P<0.05).The liver index and gallbladder index of PAMK+CTX group had no significant difference compared with the control group(P>0.05).2) The liver 8-hydroxy-deoxyguanosine content of CTX group and PAMK+CTX group was significantly higher than that of the control group(P<0.05).3) The CTX could cause liver pathological damages,DNA oxidative damage of hepatic cells and some lesions in the liver,such as tissue fibrosis,inflammatory small focus,cholestasis et al.The PAMK could alleviate extent of damage of liver tissue,reduce the number of necrotic hepatocytes.4) Compared with the control group,the liver malondialdehyde(MDA) content of CTX group was significantly increased(P<0.05),and the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT),inducible nitric oxide synthase(iNOS) in liver were significantly decreased(P<0.05).The MDA content and GSH-Px,iNOS activities in liver of PAMK+CTX group had no significant difference compared with the control group(P>0.05).5) The mRNA relative expressions of Tolllike receptor 4(TLR4),tumor necrosis factor receptor-associated factor 6(TRAF6),mitogen activated protein kinase kinase 1(MEKK1),mitogen-activated protein kinase kinase 3(MKK3),p38 mitogen-activated protein kinase(p38 MAPK),inhibitor of nuclear factor-κB α(IκBα) and inhibitor of nuclear factor kappa-B kinase α(IKKα) in liver of CTX group were significantly lower than those of the control group(P<0.05).The mRNA relative expressions of TLR4,myeloid differentiation primary response(MyD88),MEKK1,MKK3,mitogen-activated protein kinase kinase 6(MKK6),inhibitor of nuclear factor kappa-B kinase ε(IKKε) and IκBαin liver of PAMK + CTX group were significantly higher than those of CTX group(P<0.05),the mRNA relative expressions of MyD88 and MKK6 in liver were significantly higher than those of the control group(P<0.05),and the mRNA relative expressions of TRAF6,MEKK1,MKK3 and p38 MAPK in liver were significantly lower than those of the control group(P<0.05).These results suggested that PAMK may regulate liver injury induced by CTX in goslings via TLR4 signaling pathway.
引文
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[15]SZABO G,DOLGANIUC A,MANDREKAR P.Pattern recognition receptors:a contemporary view on liver diseases[J].Hepatology,2006,44(2):287-298.
[16]GUSTOT T,LEMMERS A,MORENO C,et al.Differential liver sensitization to toll-like receptor pathways in mice with alcoholic fatty liver[J].Hepatology,2006,43(5):989-1000.
[17]STRKEL P,DE SAEGER C,STRAIN A J,et al.NFκB,cytokines,TLR 3 and 7 expression in human end-stage HCV and alcoholic liver disease[J].European Journal of Clinical Investigation,2010,40(7):575-584.
[18]SCHWABE R F,SEKI E,BRENNER D A.Toll-like receptor signaling in the liver[J].Gastroenterology,2006,130(6):1886-1900.
[19]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.
[20]YOON Y D,HAN S B,KANG J S,et al.Toll-like receptor4-dependent activation of macrophages by polysaccharide isolated from the radix of Platycodon grandiflorum[J].International Immunopharmacology,2003,3(13/14):1873-1882.
[21]KIM S J,PARK H J,SHIN H J,et al.Suppression of TRIF-dependent signaling pathway of toll-like receptors by allyl isothiocyanate in RAW 264.7 macrophages[J].International Immunopharmacology,2012,13(4):403-407.
[22]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].Scientific Reports,2017,7:44822.
[23]BORAZJANI N J,TABARSA M,YOU S G,et al.Purification,molecular properties,structural characterization,a nd immunomodulatory activities of water soluble polysaccharides from Sargassum angustifolium[J].International Journal of Biological Macromolecules,2018,109:793-802.
[2]XI Q Y,JIANG Y,ZHAO S,et al.Effect of ginseng polysaccharides on the immunity and growth of piglets by dietary supplementation during late pregnancy and lactating sows[J].Animal Science Journal,2017,88(6):863-872.
[3]ZHU N,LV X C,WANG Y Y,et al.Comparison of immunoregulatory effects of polysaccharides from three natural herbs and cellular uptake in dendritic cells[J].International Journal of Biological Macromolecules,2016,93:940-951.
[4]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.
[5]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.
[6]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.
[7]XU D N,LI W Y,LI B X,et al.The effect of selenium and polysaccharide of Atractylodes macrocephala Koidz.(PAMK)on endoplasmic reticulum stress and apoptosis in chicken spleen induced by heat stress[J].RSC Advances,2017,7(13):7519-7525.
[8]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.
[9]MOIGNET A,HASANALI Z,ZAMBELLO R,et al.Cyclophosphamide as a first-line therapy in LGL leukemia[J].Leukemia,2014,28(5):1134-1136.
[10]HABIBI E,SHOKRZADEH M,CHABRA A,et al.Protective effects of Origanum vulgare ethanol extract against cyclophosphamide-induced liver toxicity in mice[J].Pharmaceutical Biology,2015,53(1):10-15.
[11]MANSOUR D F,SALEH D O,MOSTAFA R E.Genistein ameliorates cyclophosphamide-induced hepatotoxicity by modulation of oxidative stress and inflammatory mediators[J].Open Access Macedonian Journal of Medical Sciences,2017,5(7):836-843.
[12]ZHOU Y L,CHEN X Y,YI R K,et al.Immunomodulatory effect of tremella polysaccharides against cyclophosphamideinduced immunosuppression in mice[J].Molecules,2018,23(2):E239.
[13]LI Q,CHEN G Y,CHEN H,et al.Se-enriched G.frondosa polysaccharide protects against immunosuppression in cyclophosphamide-induced mice via MAPKs signal transduction pathway[J].Carbohydrate Polymers,2018,196:445-456.
[14]LIN S T,HAO G X,LONG M,et al.Oyster(Ostrea plicatula Gmelin)polysaccharides intervention ameliorates cyclophosphamide-induced genotoxicity and hepatotoxicity in mice via the Nrf2-ARE pathway[J].Biomedicine&Pharmacotherapy,2017,95:1067-1071.
[15]SZABO G,DOLGANIUC A,MANDREKAR P.Pattern recognition receptors:a contemporary view on liver diseases[J].Hepatology,2006,44(2):287-298.
[16]GUSTOT T,LEMMERS A,MORENO C,et al.Differential liver sensitization to toll-like receptor pathways in mice with alcoholic fatty liver[J].Hepatology,2006,43(5):989-1000.
[17]STRKEL P,DE SAEGER C,STRAIN A J,et al.NFκB,cytokines,TLR 3 and 7 expression in human end-stage HCV and alcoholic liver disease[J].European Journal of Clinical Investigation,2010,40(7):575-584.
[18]SCHWABE R F,SEKI E,BRENNER D A.Toll-like receptor signaling in the liver[J].Gastroenterology,2006,130(6):1886-1900.
[19]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.
[20]YOON Y D,HAN S B,KANG J S,et al.Toll-like receptor4-dependent activation of macrophages by polysaccharide isolated from the radix of Platycodon grandiflorum[J].International Immunopharmacology,2003,3(13/14):1873-1882.
[21]KIM S J,PARK H J,SHIN H J,et al.Suppression of TRIF-dependent signaling pathway of toll-like receptors by allyl isothiocyanate in RAW 264.7 macrophages[J].International Immunopharmacology,2012,13(4):403-407.
[22]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].Scientific Reports,2017,7:44822.
[23]BORAZJANI N J,TABARSA M,YOU S G,et al.Purification,molecular properties,structural characterization,a nd immunomodulatory activities of water soluble polysaccharides from Sargassum angustifolium[J].International Journal of Biological Macromolecules,2018,109:793-802.