镰刀菌毒素对断奶仔猪脾脏抗氧化能力和白细胞介素-1β、白细胞介素-6分布和表达的影响
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摘要
本试验旨在研究自然霉变饲粮中镰刀菌毒素对断奶仔猪脾脏抗氧化能力和白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)分布和表达的影响。选择35日龄平均体重为(8.45±0.94)kg的健康三元杂交(杜×长×大)断奶仔猪母猪40头,随机分为2组,每组20头。对照组饲喂基础饲粮,镰刀菌毒素组饲喂含镰刀菌毒素[玉米赤霉烯酮(ZEN)0.90 mg/kg;呕吐毒素(DON)1.43mg/kg,烟曲霉毒素(FUM)5.85 mg/kg]的试验饲粮。预试期7d,正试期35d。结果表明:1)与对照组相比,镰刀菌毒素显著降低了断奶仔猪血清和脾脏谷胱甘肽过氧化物酶(GSH-Px)和总超氧化物歧化酶(T-SOD)活性(P<0.05),显著升高了丙二醛(MDA)含量(P<0.05)。2)镰刀菌毒素使断奶仔猪脾脏白髓区明显变小,红髓区扩张且出现近圆形小空洞,动脉周围淋巴鞘中淋巴细胞数量较少。3)镰刀菌毒素导致断奶仔猪脾脏IL-1β和IL-6阳性细胞主要集中于白髓边缘,且靠近血窦的地方阳性点更多。4)与对照组相比,镰刀菌毒素显著升高了断奶仔猪脾脏IL-1β和IL-6mRNA相对表达量(P<0.05)。由此可见,饲粮中镰刀菌毒素显著影响断奶仔猪血清和脾脏抗氧化能力,并通过改变脾脏IL-1β和IL-6的分布和表达,降低脾脏的免疫功能。
The aims of the present study were to investigate the effects of Fusarium toxins in naturally contaminated diet on antioxidant capacity and distribution and expression of interleukin-1β(IL-1β)and interleukin-6(IL-6)in spleen of weaned piglets.Forty healthy female weaned piglets(Duroc×Landrace×Yorkshire)aged at 35 days with an average body weight of(8.45±0.94)kg were randomly allocated into 2groups with 20 piglets per group.Piglets in control group were fed a basal diet,and the others in Fusarium toxins group were fed Fusarium toxins[0.90 mg/kg zearalenone(ZEN);1.43 mg/kg deoxynivalenol(DON);5.85 mg/kg fumonisin(FUM)]contaminated experimental diet.The experiment lasted for 35 days after 7days adaptation.The results showed as follows:1)compared with the control group,Fusarium toxins significantly decreased the activities of glutathione peroxidase(GSH-Px)and total superoxide dismutase(T-SOD)in serum and spleen of weaned piglets(P<0.05),whereas significantly increased the content of malondialdehyde(MDA)in serum and spleen(P<0.05).2)Fusarium toxins significantly reduced the white pulp zone in spleen of weaned piglets,enlarged the red pulp zone and appeared round small cavity,while only a few lymphocytes were seen in the peripheral lymphatic sheath.3)The positive cells of IL-1βand IL-6in spleen of weaned piglets were mainly distributed in the edge of the white pulp,and concentrated area of positive cells were found near the blood sinus in spleen after exposure to Fusariumtoxins.4)Compared with the control group,Fusarium toxins significantly increased the relative expression of IL-1βand IL-6mRNA in spleen of weaned piglets(P<0.05).It is suggested that dietary Fusariumtoxins has significant effects on antioxidant capacity in serum and spleen,and decreases the immune function in spleen by changing distribution and expression of IL-1βand IL-6in spleen of weaned piglets.
The aims of the present study were to investigate the effects of Fusarium toxins in naturally contaminated diet on antioxidant capacity and distribution and expression of interleukin-1β(IL-1β)and interleukin-6(IL-6)in spleen of weaned piglets.Forty healthy female weaned piglets(Duroc×Landrace×Yorkshire)aged at 35 days with an average body weight of(8.45±0.94)kg were randomly allocated into 2groups with 20 piglets per group.Piglets in control group were fed a basal diet,and the others in Fusarium toxins group were fed Fusarium toxins[0.90 mg/kg zearalenone(ZEN);1.43 mg/kg deoxynivalenol(DON);5.85 mg/kg fumonisin(FUM)]contaminated experimental diet.The experiment lasted for 35 days after 7days adaptation.The results showed as follows:1)compared with the control group,Fusarium toxins significantly decreased the activities of glutathione peroxidase(GSH-Px)and total superoxide dismutase(T-SOD)in serum and spleen of weaned piglets(P<0.05),whereas significantly increased the content of malondialdehyde(MDA)in serum and spleen(P<0.05).2)Fusarium toxins significantly reduced the white pulp zone in spleen of weaned piglets,enlarged the red pulp zone and appeared round small cavity,while only a few lymphocytes were seen in the peripheral lymphatic sheath.3)The positive cells of IL-1βand IL-6in spleen of weaned piglets were mainly distributed in the edge of the white pulp,and concentrated area of positive cells were found near the blood sinus in spleen after exposure to Fusariumtoxins.4)Compared with the control group,Fusarium toxins significantly increased the relative expression of IL-1βand IL-6mRNA in spleen of weaned piglets(P<0.05).It is suggested that dietary Fusariumtoxins has significant effects on antioxidant capacity in serum and spleen,and decreases the immune function in spleen by changing distribution and expression of IL-1βand IL-6in spleen of weaned piglets.
引文
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[6]WAN L Y M,WOO C S J,TURNER P C,et al.Individual and combined effects of Fusarium toxins on the mRNA expression of pro-inflammatory cytokines in swine jejunal epithelial cells[J].Toxicology Letters,2013,220(3):238-246.
[7]National Research Council.Nutrient requirements of swine[M].11th ed.Washington D.C.:National Academy Press,2012.
[8]张丽英.饲料分析及饲料质量检测技术[M].2版.北京:中国农业大学出版社,2003.
[9]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J].Methods,2001,25(4):402-408.
[10]陈祥兴,杨维仁,张崇玉,等.镰刀菌毒素对断奶仔猪生长性能、小肠二糖酶活性和抗氧化能力的影响[J].动物营养学报,2015,27(6):1875-1882.
[11]中华人民共和国国家质量监督检验检疫总局.GB13078.2—2006饲料卫生标准饲料中赭曲霉毒素A和玉米赤霉烯酮的允许量[S].北京:中国标准出版社,2006.
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[13]European Commission.Commission recommendation of 17August 2006on the presence of deoxynivalenol,zearalenone,ochratoxin A,T-2and HT-2and fumonisins in products intended for animal feeding[S].Official Journal of European Union,2006,229:7-9.
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[15]JIANG S Z,YANG Z B,YANG W R,et al.Effects of purified zearalenone on growth performance,organ size,serum metabolites,and oxidative stress in postweaning gilts[J].Journal of Animal Science,2011,89(10):3008-3015.
[16]蒋竹英,李丽立,唐利华,等.DON污染饲粮添加竹炭和竹醋液对断奶仔猪抗氧化性能及小肠黏膜形态的影响[J].西北农林科技大学学报:自然科学版,2013,41(12):37-42.
[17]闫俊书,单安山,王环宇.五味子提取物对AA肉仔鸡抗氧化功能的影响[J].中国畜牧杂志,2008,44(17):33-37.
[18]CHEUNG C C C,ZHENG G J,LI A M Y,et al.Relationships between tissue concentrations of polycyclic aromatic hydrocarbons and antioxidative responses of marine mussels,Perna viridis[J].Aquatic Toxicology,2001,52(3/4):189-203.
[19]DRGE W.Free radicals in the physiological control of cell function[J].Physiological Reviews,2002,82(1):47-95.
[20]KOUADIO J H,MOBIO T A,BAUDRIMONT I,et al.Comparative study of cytotoxicity and oxidative stress induced by deoxynivalenol,zearalenone or fumonisin B1in human intestinal cell line Caco-2[J].Toxicology,2005,213(1/2):56-65.
[21]苏军.镰刀菌毒素对猪的抗营养效应及其机制研究[D].博士论学位文.雅安:四川农业大学,2008.
[22]MCLEAN M.The phytotoxicity of Fusarium metabolites:an update since 1989[J].Mycopathologia,1996,133(3):163-179.
[23]JIANG S Z,LI Z,WANG G Y,et al.Effects of Fusarium mycotoxins with yeast cell wall absorbent on hematology,serum biochemistry,and oxidative stress in broiler chickens[J].The Journal of Applied Poultry Research,2014,23(2):165-173.
[24]ABBS S,SALAH-ABBS J B,OUANES Z,et al.Preventive role of phyllosilicate clay on the immunological and biochemical toxicity of zearalenone in Balb/c mice[J].International Immunopharmacology,2006,6(8):1251-1258.
[25]FORSELL J H,WITT M F,TAI J H,et al.Effects of8-week exposure of the B6C3F1mouse to dietary deoxynivalenol(vomitoxin)and zearalenone[J].Food and Chemical Toxicology,1986,24(3):213-219.
[26]DONG M,HE X J,TULAYAKUL P,et al.The toxic effects and fate of intravenously administered zearalenone in goats[J].Toxicon,2010,55(2/3):523-530.
[27]王凯.IL-6在炎症性肠病大鼠脑、结肠组织中表达及信号转导机制研究[D].长春:吉林大学,2010.
[28]SABA A A,GODZIACHVILI V,MAVANI A K,et al.Serum levels of interleukin 1and tumor necrosis factor alpha correlate with peritoneal adhesion grades in humans after major abdominal surgery[J].The American Surgeon,1998,64(8):734-737.
[29]余传霖.细菌内毒素的研究现状[J].国外医学:微生物学分册,1997(5):29-31.
[30]俞晓峰.细菌内毒素作用机理的研究进展[J].国外医学:微生物学分册,1993(5):217-219,216.
[31]PAPANICOLAOU D A,WILDER R L,MANOLAGAS S C,et al.The pathophysiologic roles of interleukin-6in human disease[J].Annals of Internal Medicine,1998,128(2):127-137.
[32]VLAHAKOS D,ARKADOPOULOS N,KOSTOPANAGIOTOU G,et al.Deferoxamine attenuates lipid peroxidation,blocks interleukin-6production,ameliorates sepsis inflammatory response syndrome,and confers renoprotection after acute hepatic ischemia in pigs[J].Artificial Organs,2012,36(4):400-408.
[33]雷明彦.饲料中常见霉菌毒素对小鼠的联合毒性作用及机制的研究[D].博士学位论文.武汉:华中农业大学,2013.
[34]GRENIER B,LOUREIRO-BRACARENSE A P,LUCIOLI J,et al.Individual and combined effects of subclinical doses of deoxynivalenol and fumonisins in piglets[J].Molecular Nutrition&Food Research,2011,55(5):761-771.
[35]LI Z,YANG Z B,YANG W R,et al.Effects of feedborne Fusarium mycotoxins with or without yeast cell wall adsorbent on organ weight,serum biochemistry,and immunological parameters of broiler chickens[J].Poultry Science,2012,91(10):2487-2495.
[36]GIRISH C,SMITH T.Impact of feed-borne mycotoxins on avian cell-mediated and humoral immune responses[J].World Mycotoxin Journal,2008,1(2):105-121.
[2]牛群升,杨维仁,黄丽波,等.镰刀菌毒素对断奶小母猪阴户、生殖器官指数、子宫雌激素受体分布和表达的影响[J].动物营养学报,2016,28(5):1525-1533.
[3]HUSSEIN H S,BRASEL J M.Toxicity,metabolism,and impact of mycotoxins on humans and animals[J].Toxicology,2001,167(2):101-134.
[4]GEREZ J R,PINTON P,CALLU P,et al.Deoxynivalenol alone or in combination with nivalenol and zearalenone induce systemic histological changes in pigs[J].Experimental and Toxicologic Pathology,2015,67(2):89-98.
[5]WAN L Y M,TURNER P C,EL-NEZAMI H.Individual and combined cytotoxic effects of Fusarium toxins(deoxynivalenol,nivalenol,zearalenone and fumonisins B1)on swine jejunal epithelial cells[J].Food and Chemical Toxicology,2013,57:276-283.
[6]WAN L Y M,WOO C S J,TURNER P C,et al.Individual and combined effects of Fusarium toxins on the mRNA expression of pro-inflammatory cytokines in swine jejunal epithelial cells[J].Toxicology Letters,2013,220(3):238-246.
[7]National Research Council.Nutrient requirements of swine[M].11th ed.Washington D.C.:National Academy Press,2012.
[8]张丽英.饲料分析及饲料质量检测技术[M].2版.北京:中国农业大学出版社,2003.
[9]LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method[J].Methods,2001,25(4):402-408.
[10]陈祥兴,杨维仁,张崇玉,等.镰刀菌毒素对断奶仔猪生长性能、小肠二糖酶活性和抗氧化能力的影响[J].动物营养学报,2015,27(6):1875-1882.
[11]中华人民共和国国家质量监督检验检疫总局.GB13078.2—2006饲料卫生标准饲料中赭曲霉毒素A和玉米赤霉烯酮的允许量[S].北京:中国标准出版社,2006.
[12]中华人民共和国国家质量监督检验检疫总局.GB13078.3—2007配合饲料中脱氧雪腐镰刀菌烯醇的允许量[S].北京:中国标准出版社,2007.
[13]European Commission.Commission recommendation of 17August 2006on the presence of deoxynivalenol,zearalenone,ochratoxin A,T-2and HT-2and fumonisins in products intended for animal feeding[S].Official Journal of European Union,2006,229:7-9.
[14]SALAH-ABBS J B,ABBS S,OUANES Z,et al.Tunisian radish extract(Raphanus sativus)enhances the antioxidant status and protects against oxidative stress induced by zearalenone in Balb/c mice[J].Journal of Applied Toxicology,2008,28(1):6-14.
[15]JIANG S Z,YANG Z B,YANG W R,et al.Effects of purified zearalenone on growth performance,organ size,serum metabolites,and oxidative stress in postweaning gilts[J].Journal of Animal Science,2011,89(10):3008-3015.
[16]蒋竹英,李丽立,唐利华,等.DON污染饲粮添加竹炭和竹醋液对断奶仔猪抗氧化性能及小肠黏膜形态的影响[J].西北农林科技大学学报:自然科学版,2013,41(12):37-42.
[17]闫俊书,单安山,王环宇.五味子提取物对AA肉仔鸡抗氧化功能的影响[J].中国畜牧杂志,2008,44(17):33-37.
[18]CHEUNG C C C,ZHENG G J,LI A M Y,et al.Relationships between tissue concentrations of polycyclic aromatic hydrocarbons and antioxidative responses of marine mussels,Perna viridis[J].Aquatic Toxicology,2001,52(3/4):189-203.
[19]DRGE W.Free radicals in the physiological control of cell function[J].Physiological Reviews,2002,82(1):47-95.
[20]KOUADIO J H,MOBIO T A,BAUDRIMONT I,et al.Comparative study of cytotoxicity and oxidative stress induced by deoxynivalenol,zearalenone or fumonisin B1in human intestinal cell line Caco-2[J].Toxicology,2005,213(1/2):56-65.
[21]苏军.镰刀菌毒素对猪的抗营养效应及其机制研究[D].博士论学位文.雅安:四川农业大学,2008.
[22]MCLEAN M.The phytotoxicity of Fusarium metabolites:an update since 1989[J].Mycopathologia,1996,133(3):163-179.
[23]JIANG S Z,LI Z,WANG G Y,et al.Effects of Fusarium mycotoxins with yeast cell wall absorbent on hematology,serum biochemistry,and oxidative stress in broiler chickens[J].The Journal of Applied Poultry Research,2014,23(2):165-173.
[24]ABBS S,SALAH-ABBS J B,OUANES Z,et al.Preventive role of phyllosilicate clay on the immunological and biochemical toxicity of zearalenone in Balb/c mice[J].International Immunopharmacology,2006,6(8):1251-1258.
[25]FORSELL J H,WITT M F,TAI J H,et al.Effects of8-week exposure of the B6C3F1mouse to dietary deoxynivalenol(vomitoxin)and zearalenone[J].Food and Chemical Toxicology,1986,24(3):213-219.
[26]DONG M,HE X J,TULAYAKUL P,et al.The toxic effects and fate of intravenously administered zearalenone in goats[J].Toxicon,2010,55(2/3):523-530.
[27]王凯.IL-6在炎症性肠病大鼠脑、结肠组织中表达及信号转导机制研究[D].长春:吉林大学,2010.
[28]SABA A A,GODZIACHVILI V,MAVANI A K,et al.Serum levels of interleukin 1and tumor necrosis factor alpha correlate with peritoneal adhesion grades in humans after major abdominal surgery[J].The American Surgeon,1998,64(8):734-737.
[29]余传霖.细菌内毒素的研究现状[J].国外医学:微生物学分册,1997(5):29-31.
[30]俞晓峰.细菌内毒素作用机理的研究进展[J].国外医学:微生物学分册,1993(5):217-219,216.
[31]PAPANICOLAOU D A,WILDER R L,MANOLAGAS S C,et al.The pathophysiologic roles of interleukin-6in human disease[J].Annals of Internal Medicine,1998,128(2):127-137.
[32]VLAHAKOS D,ARKADOPOULOS N,KOSTOPANAGIOTOU G,et al.Deferoxamine attenuates lipid peroxidation,blocks interleukin-6production,ameliorates sepsis inflammatory response syndrome,and confers renoprotection after acute hepatic ischemia in pigs[J].Artificial Organs,2012,36(4):400-408.
[33]雷明彦.饲料中常见霉菌毒素对小鼠的联合毒性作用及机制的研究[D].博士学位论文.武汉:华中农业大学,2013.
[34]GRENIER B,LOUREIRO-BRACARENSE A P,LUCIOLI J,et al.Individual and combined effects of subclinical doses of deoxynivalenol and fumonisins in piglets[J].Molecular Nutrition&Food Research,2011,55(5):761-771.
[35]LI Z,YANG Z B,YANG W R,et al.Effects of feedborne Fusarium mycotoxins with or without yeast cell wall adsorbent on organ weight,serum biochemistry,and immunological parameters of broiler chickens[J].Poultry Science,2012,91(10):2487-2495.
[36]GIRISH C,SMITH T.Impact of feed-borne mycotoxins on avian cell-mediated and humoral immune responses[J].World Mycotoxin Journal,2008,1(2):105-121.