硒化大蒜多糖对环磷酰胺所致免疫抑制鸡的颉颃作用
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摘要
为探究硒化大蒜多糖对环磷酰胺所致免疫抑制的颉颃作用,将336只11日龄非免疫健康罗曼雏鸡随机均分为7组:空白对照组(BC)、免疫对照组(VC)、环磷酰胺对照组(Cy)、sGPS3、sGPS5、sGPS6和药物对照组(APS),每组6个重复,每个重复8只鸡。BC组和VC组注射0.5mL生理盐水,其余组均肌肉注射8mg/mL环磷酰胺0.5mL,每天1次,连续3d;14日龄时,除BC组外,其余组均用新城疫疫苗免疫,同时3个硒大蒜化多糖组分别注射0.5mL浓度为1mg/mL的sGPS3、sGPS5、sGPS6,APS组肌肉注射黄芪多糖注射液0.5mL,Cy组、VC组和BC组注射生理盐水0.5mL,每天1次,连续3d,28日龄二免。分别于首免后的第7、14、21、28天,每组随机抽取6只罗曼鸡翼静脉采血,分离血清,β-微量法检测血清ND-HI抗体效价、ELISA法检测血清γ干扰素(IFN-γ)和白细胞介素2(IL-2)含量。称量体重后采集胸腺、脾脏和法氏囊组织并称重,计算免疫器官指数。结果显示,各给药组的血清ND-HI抗体效价、血清IFN-γ和IL-2含量、法氏囊指数均显著高于Cy组(P<0.05);sGPS6组血清抗体效价、IFN-γ含量在28d,血清IL-2含量在21、28d均显著高于其余各组(P<0.05);在免疫后21、28d,4个给药组中sGPS6组体重、法氏囊指数最高,显著高于其余3个给药组(P<0.05);在免疫后28d,4个给药组中sGPS6组胸腺指数、脾脏指数最高,显著高于其余3个给药组(P<0.05)。结果表明,硒化大蒜多糖对环磷酰胺所致的免疫抑制具有颉颃作用,其中sGPS6组的活性最高,可作为新型免疫抑制颉颃剂的候选药。
In order to observe the antagonistic effect of selenizing garlic polysaccharide on immunosuppressed chicken induced by cyclophosphamide,336 11-day-old roman chickens were randomly divided into seven groups:Blank control group(BC),vaccine group(VC),cyclophosphamidegroup(Cy),sGPS3 group,sGPS5 group,sGPS6 group and drug control group(APS),with 6 duplications per group and 8 chickens per duplication.Each chicken except that in BC and VC groups were intramuscularly injected with 8 mg/mL cyclophosphamide 0.5 mL,while each chicken in BC and VC groups were injected with the same amount of saline,once a day for three successive days.At the 14 dof age,The chickens except BC group were vaccinated with ND vaccine,repeated vaccination at 28 days old.At the first vaccination,the chickens in three sGPSs groups were injected with 0.5 mL of 1 mg/mL sGPS3,sGPS5 and sGPS6,respectively.The chickens in APS group were intramuscularly injected with 0.5 mL Astragalus polysaccharide,while that in Cy,VC and BC groups were injected with the same amount of saline,once a day for three successive days.Six chickens were selected randomly from each group for collecting wing venous blood on 7,14,21 and 28 days after the first vaccination,and the serum ND-HI antibody titer were determined byβ-micro method,IFN-γand IL-2 contents were determined by ELISA.The body weighed,thymus,spleen,bursa of Fabricius were collected after culling,and the immune organ indexes were calculated.The result showed that the serum ND-HI antibody titer,IFN-γand IL-2 contents and bursa of Fabricius index of each administration groups were higher than that of the Cy group(P<0.05);the serum ND-HI antibody titer and IFN-γcontents at 21 d,IL-2 content at 21 and 28 din sGPS6 group were significantly higher than that of other groups(P<0.05);At 21 and 28 dafter first vaccination,the body weight and bursa of Fabricius index in sGPS6 group were significantly higher than that of other three administration groups(P<0.05).At 28 dafter first vaccination,the thymus and spleen indexes in sGPS6 group were significantly higher than that of other three administration groups(P<0.05).In conlusion,the selenizing garlic polysaccharide had the antagonistic effect on immunosuppressed chicken induced by cyclophosphamide,and sGPS6 had the better effect which could be used as candidate drug for novel immunosuppressive antagonists.
In order to observe the antagonistic effect of selenizing garlic polysaccharide on immunosuppressed chicken induced by cyclophosphamide,336 11-day-old roman chickens were randomly divided into seven groups:Blank control group(BC),vaccine group(VC),cyclophosphamidegroup(Cy),sGPS3 group,sGPS5 group,sGPS6 group and drug control group(APS),with 6 duplications per group and 8 chickens per duplication.Each chicken except that in BC and VC groups were intramuscularly injected with 8 mg/mL cyclophosphamide 0.5 mL,while each chicken in BC and VC groups were injected with the same amount of saline,once a day for three successive days.At the 14 dof age,The chickens except BC group were vaccinated with ND vaccine,repeated vaccination at 28 days old.At the first vaccination,the chickens in three sGPSs groups were injected with 0.5 mL of 1 mg/mL sGPS3,sGPS5 and sGPS6,respectively.The chickens in APS group were intramuscularly injected with 0.5 mL Astragalus polysaccharide,while that in Cy,VC and BC groups were injected with the same amount of saline,once a day for three successive days.Six chickens were selected randomly from each group for collecting wing venous blood on 7,14,21 and 28 days after the first vaccination,and the serum ND-HI antibody titer were determined byβ-micro method,IFN-γand IL-2 contents were determined by ELISA.The body weighed,thymus,spleen,bursa of Fabricius were collected after culling,and the immune organ indexes were calculated.The result showed that the serum ND-HI antibody titer,IFN-γand IL-2 contents and bursa of Fabricius index of each administration groups were higher than that of the Cy group(P<0.05);the serum ND-HI antibody titer and IFN-γcontents at 21 d,IL-2 content at 21 and 28 din sGPS6 group were significantly higher than that of other groups(P<0.05);At 21 and 28 dafter first vaccination,the body weight and bursa of Fabricius index in sGPS6 group were significantly higher than that of other three administration groups(P<0.05).At 28 dafter first vaccination,the thymus and spleen indexes in sGPS6 group were significantly higher than that of other three administration groups(P<0.05).In conlusion,the selenizing garlic polysaccharide had the antagonistic effect on immunosuppressed chicken induced by cyclophosphamide,and sGPS6 had the better effect which could be used as candidate drug for novel immunosuppressive antagonists.
引文
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[7]贾宁,王汉,郑晶.复方党参提取物对环磷酰胺处理小鼠免疫功能的调节作用[J].中国实验方剂学,2011,9(17):206-209.JIA N,WANG H,ZHENG J.Immunomodulative effect of compound codonopsis radix extract on immunosuppressive mice induced by cyclophosphamide[J].Chinese Journal of Experimental Traditional Medical Formulae,2011,9(17):206-209.(in Chinese)
[8]郑敏,潘世斌,姜有定,等.大蒜多糖对肝损伤小鼠血清和肝组织ALT、AST的影响程度及急性毒性试验[J].咸宁学院学报,2003,17(2):85-87.ZHENG M,PAN S B,JIANG Y D,et al.An acute toxicity testing of Allium sativum L.polysaccharide and its effects on the ALT and AST in injured mice serum or liver[J].Journal of Xianning College,2003,17(2):85-87.(in Chinese)
[9] MA X,GUO Z,WANG D,et al.Effects of sulfated polysaccha-rides and their prescriptions on immune response of ND vaccine in chickens[J].Carbohydrate Polymers,2010,82:9-13.
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[12]李晓坤,闫吉昌,崔晓莹,等.硒多糖的研究与应用进展[J].中草药,2006,37(7):7-8.LI X K,YAN J C,CUI X Y,et al.Research progress and application of selenium polysaccharides[J].Chinese Traditional and Herbal Medicine,2006,37(7):7-8.(in Chinese)
[13]杨晶晶,黄家锟,蒋开年,等.正交设计法优化红景天多糖硒制备工艺及其性质研究[J].中成药,2010,32(6):1065-1067.YANG J J,HUANG J K,JIANG K N,et al.The research of optimization process for selenizing Rhodiola polysaccharides according to orthogonal test and its character[J].Chinese Traditional Patent Medicine,2010,32(6):1065-1067.(in Chinese)
[14]邱树磊.硒化大蒜多糖和硒化枸杞多糖的增强免疫和抗氧化活性研究[D].南京:南京农业大学,2014.QIU S L.The immune-enhancing and antioxidative activities of selenizing garlic polysaccharide and selenizing lyxium polysaccharide[D].Nanjing:Nanjing Agricultural University,2014.(in Chinese)
[15]郭利伟.中药多糖复方免疫増强剂AEI及其作用机理研究[D].南京:南京农业大学,2012.GUO L W.Studies on compound Chinese herbal medicinal polysaccharide immunopotentiator AEI and its action mechanism[D].Nanjing:Nanjing Agricultural University,2012.(in Chinese)
[16] WANG H,ZHANG D,ZHANG L,et al.The research of preparation and immunology of selenizing Ganoderma lucidum polysaccharides[J].Journal of Food Science,2009,30(17):316-318.
[17] QIN T,CHEN J,WANG D,et al.Optimization of selenylation conditions for Chinese angelica polysaccharide based on immune-enhancing activity[J].Carbohydrate Polymers,2013,92(1):645-650.
[18] GUO L,WANG D Y,HU Y L,et al.Adjuvanticity of compound polysaccharides on chickens against Newcastle disease and avian influenza vaccine[J].International Journal of Biological Macromolecules,2012,50(3):512-517.
[19] FAN Y,HU Y,WANG D,et al.Epimedium polysaccharide and propolis flavone can synergistically stimulate lymphocyte proliferation in vitro and enhance the immune responses to ND vaccine in chickens[J].International Journal of Biological Macromolecules,2010,47(2):87-92.
[20]周建强,潘琦,王涛,等.党参多糖对免疫雏鸡抗体效价和淋巴细胞增殖的影响[J].江苏农业科学,2010,3:263-264.ZHOU J Q,PAN Q,WANG T,et al.Effects of Codonopsis pilosula polysaccharides on antibody titer and lymphocyte proliferation in chickens[J].Jiangsu Agricultural Sciences,2010,3:263-264.(in Chinese)
[21]周泽伟,李楠,余峥嵘,等.黄芪灵芝口服液对免疫新城疫疫苗雏鸡免疫功能的影响[J].中国畜牧兽医,2015,42(7):1859-1864.ZHOU Z W,LI N,YU Z R,et al.Effect of the oral liquid of Astragalus and Lucidumpolysaccharides on immunological function in chichens immuned with ND vaccine[J].China Animal Husbandry&Veterinary Medicine,2015,42(7):1859-1864.(in Chinese)
[22] FAN Y,LU Y,WANG D,et al.Effect of epimedium polysaccharide-propolis flavone immunopotentiator on immunosuppression induced by cyclophosphamide in chickens[J].Cellular Immunology,2013,281(1):37-43.
[23] THAKUR A,PEDERSEN L E,JUNGERSEN G.Immune markers and correlates of protection for vaccine induced immune responses[J].Vaccine,2012,30(33):4907-4920.
[24]于晓东,王立辛.连翘苷拮抗环磷酰胺所致小鼠免疫抑制的实验研究[J].中国免疫学杂志,2017,33(8):1177-1180.YU X D,WANG L X.Experimental study on phillyrin resist mice immunosuppression induced by cyclophosphamide[J].Chinese Journal of Immunology,2017,33(8):1177-1180.(in Chinese)
[25]杨汉春.动物免疫学[M].北京:中国农业出版社,2002.YANG H C.Animal Immunology[M].Beijing:China Agriculture Press,2002.(in Chinese)
[26]陈兴颖.中药多糖免疫增强剂芪藿饮及作用机理研究[D].南京:南京农业大学,2014.CHEN X Y.Studies on Chinese herbal medicinal polysaccharide immunopotentiator AEO and its mechanism[D].Nanjing:Nanjing Agricultural University,2014.(in Chinese)
[27]宋玉龙,丘富安,吴秀钦,等.硒化修饰太子参多糖对免疫损伤小鼠的免疫保护作用[J].中国兽医学报,2017,11(37):2163-2180.SONG Y L,QIU F A,WU X Q,et al.Immunoprotection effect of selenium radix Pseudostellaria polysaccharide on immune injured mice[J].Chinese Journal of Veterinary Science,2017,11(37):2163-2180.(in Chinese)
[28]林丹丹,秦韬,任喆,等.硒化党参多糖对免疫抑制小鼠免疫功能的影响[J].中国畜牧兽医,2016,43(6):1544-1549.LIN D D,QIN T,REN Z,et al.Effects of selenium Codonopsis pilosula polysaccharide on immune function of immunosuppressed mice[J].China Animal Husbandry&Veterinary Medicine,2016,43(6):1544-1549.(in Chinese)
[2]高玉龙,秦立廷,王笑梅,等.禽病毒性免疫抑制病流行特点与防控对策[J].中国家禽,2012,34(15):5-11.GAO Y L,QIN L T,WANG X M,et al.The epidemic characteristics and prevention,control countermeasures of poultry viral immunosuppression diseases[J].China Poultry,2012,34(15):5-11.(in Chinese)
[3]武艳,孙莉.毛皮动物免疫抑制的危害及防控措施[J].中国畜牧兽医文摘,2013,29(3):119.WU Y,SUN L.The harm and control measures of immunosuppression in fur animals[J].Chinese Animal Husbandry and Veterinary Abstracts,2013,29(3):119.(in Chinese)
[4]周泽伟,刘浩,王凯,等.黄芪灵芝口服液对雏鸡免疫功能的影响[J].中国兽医杂志,2016,8(52):59-61.ZHOU Z W,LIU H,WANG K,et al.The effect of Astragalus ganoderma oral liquid on immune function of chicken[J].Chinese Journal of Veterinary Medicine,2016,8(52):59-61.(in Chinese)
[5]蒋蕊,考书娟,徐小娟,等.槲皮素对免疫低下小鼠体液免疫功能的影响[J].青岛农业大学学报(自然科学版),2011,28(2):118-120.JIANG R,KAO S J,XU X J,et al.Effect of quercetin on humoral immunity function in immunosuppressive mice[J].Journal of Qingdao Agricultural University(Natural Science),2011,28(2):118-120.(in Chinese)
[6]赵雅.全真一气汤复方多糖对免疫抑制小鼠免疫功能影响初探[D].广州:广州医科大学,2014.ZHAO Y.Preliminary research on Quanzhen Yiqi decoction polysaccharide for the Immune function of immunosuppressed mice[D].Guangzhou:Guangzhou Medical University,2014.(in Chinese)
[7]贾宁,王汉,郑晶.复方党参提取物对环磷酰胺处理小鼠免疫功能的调节作用[J].中国实验方剂学,2011,9(17):206-209.JIA N,WANG H,ZHENG J.Immunomodulative effect of compound codonopsis radix extract on immunosuppressive mice induced by cyclophosphamide[J].Chinese Journal of Experimental Traditional Medical Formulae,2011,9(17):206-209.(in Chinese)
[8]郑敏,潘世斌,姜有定,等.大蒜多糖对肝损伤小鼠血清和肝组织ALT、AST的影响程度及急性毒性试验[J].咸宁学院学报,2003,17(2):85-87.ZHENG M,PAN S B,JIANG Y D,et al.An acute toxicity testing of Allium sativum L.polysaccharide and its effects on the ALT and AST in injured mice serum or liver[J].Journal of Xianning College,2003,17(2):85-87.(in Chinese)
[9] MA X,GUO Z,WANG D,et al.Effects of sulfated polysaccha-rides and their prescriptions on immune response of ND vaccine in chickens[J].Carbohydrate Polymers,2010,82:9-13.
[10]胡群宝,郭宝江.螺旋藻硒多糖对小鼠免疫功能的影响[J].中国海洋药物,2001,5:18-20.HU Q B,GUO B J.Analysis of spirulina-selenium polysaccharide improvement on mice immunological competence[J].Journal of Chinese Marine Drugs,2001,5:18-20.(in Chinese)
[11] STAAF M,YANG Z N,HUTTUNEN E,et al.Structural elucidation of the viscous exopolysaccharide produced by Lactobacillus helveticus Lb161s[J].Carbohydrate Research,2000,326(2):113-119.
[12]李晓坤,闫吉昌,崔晓莹,等.硒多糖的研究与应用进展[J].中草药,2006,37(7):7-8.LI X K,YAN J C,CUI X Y,et al.Research progress and application of selenium polysaccharides[J].Chinese Traditional and Herbal Medicine,2006,37(7):7-8.(in Chinese)
[13]杨晶晶,黄家锟,蒋开年,等.正交设计法优化红景天多糖硒制备工艺及其性质研究[J].中成药,2010,32(6):1065-1067.YANG J J,HUANG J K,JIANG K N,et al.The research of optimization process for selenizing Rhodiola polysaccharides according to orthogonal test and its character[J].Chinese Traditional Patent Medicine,2010,32(6):1065-1067.(in Chinese)
[14]邱树磊.硒化大蒜多糖和硒化枸杞多糖的增强免疫和抗氧化活性研究[D].南京:南京农业大学,2014.QIU S L.The immune-enhancing and antioxidative activities of selenizing garlic polysaccharide and selenizing lyxium polysaccharide[D].Nanjing:Nanjing Agricultural University,2014.(in Chinese)
[15]郭利伟.中药多糖复方免疫増强剂AEI及其作用机理研究[D].南京:南京农业大学,2012.GUO L W.Studies on compound Chinese herbal medicinal polysaccharide immunopotentiator AEI and its action mechanism[D].Nanjing:Nanjing Agricultural University,2012.(in Chinese)
[16] WANG H,ZHANG D,ZHANG L,et al.The research of preparation and immunology of selenizing Ganoderma lucidum polysaccharides[J].Journal of Food Science,2009,30(17):316-318.
[17] QIN T,CHEN J,WANG D,et al.Optimization of selenylation conditions for Chinese angelica polysaccharide based on immune-enhancing activity[J].Carbohydrate Polymers,2013,92(1):645-650.
[18] GUO L,WANG D Y,HU Y L,et al.Adjuvanticity of compound polysaccharides on chickens against Newcastle disease and avian influenza vaccine[J].International Journal of Biological Macromolecules,2012,50(3):512-517.
[19] FAN Y,HU Y,WANG D,et al.Epimedium polysaccharide and propolis flavone can synergistically stimulate lymphocyte proliferation in vitro and enhance the immune responses to ND vaccine in chickens[J].International Journal of Biological Macromolecules,2010,47(2):87-92.
[20]周建强,潘琦,王涛,等.党参多糖对免疫雏鸡抗体效价和淋巴细胞增殖的影响[J].江苏农业科学,2010,3:263-264.ZHOU J Q,PAN Q,WANG T,et al.Effects of Codonopsis pilosula polysaccharides on antibody titer and lymphocyte proliferation in chickens[J].Jiangsu Agricultural Sciences,2010,3:263-264.(in Chinese)
[21]周泽伟,李楠,余峥嵘,等.黄芪灵芝口服液对免疫新城疫疫苗雏鸡免疫功能的影响[J].中国畜牧兽医,2015,42(7):1859-1864.ZHOU Z W,LI N,YU Z R,et al.Effect of the oral liquid of Astragalus and Lucidumpolysaccharides on immunological function in chichens immuned with ND vaccine[J].China Animal Husbandry&Veterinary Medicine,2015,42(7):1859-1864.(in Chinese)
[22] FAN Y,LU Y,WANG D,et al.Effect of epimedium polysaccharide-propolis flavone immunopotentiator on immunosuppression induced by cyclophosphamide in chickens[J].Cellular Immunology,2013,281(1):37-43.
[23] THAKUR A,PEDERSEN L E,JUNGERSEN G.Immune markers and correlates of protection for vaccine induced immune responses[J].Vaccine,2012,30(33):4907-4920.
[24]于晓东,王立辛.连翘苷拮抗环磷酰胺所致小鼠免疫抑制的实验研究[J].中国免疫学杂志,2017,33(8):1177-1180.YU X D,WANG L X.Experimental study on phillyrin resist mice immunosuppression induced by cyclophosphamide[J].Chinese Journal of Immunology,2017,33(8):1177-1180.(in Chinese)
[25]杨汉春.动物免疫学[M].北京:中国农业出版社,2002.YANG H C.Animal Immunology[M].Beijing:China Agriculture Press,2002.(in Chinese)
[26]陈兴颖.中药多糖免疫增强剂芪藿饮及作用机理研究[D].南京:南京农业大学,2014.CHEN X Y.Studies on Chinese herbal medicinal polysaccharide immunopotentiator AEO and its mechanism[D].Nanjing:Nanjing Agricultural University,2014.(in Chinese)
[27]宋玉龙,丘富安,吴秀钦,等.硒化修饰太子参多糖对免疫损伤小鼠的免疫保护作用[J].中国兽医学报,2017,11(37):2163-2180.SONG Y L,QIU F A,WU X Q,et al.Immunoprotection effect of selenium radix Pseudostellaria polysaccharide on immune injured mice[J].Chinese Journal of Veterinary Science,2017,11(37):2163-2180.(in Chinese)
[28]林丹丹,秦韬,任喆,等.硒化党参多糖对免疫抑制小鼠免疫功能的影响[J].中国畜牧兽医,2016,43(6):1544-1549.LIN D D,QIN T,REN Z,et al.Effects of selenium Codonopsis pilosula polysaccharide on immune function of immunosuppressed mice[J].China Animal Husbandry&Veterinary Medicine,2016,43(6):1544-1549.(in Chinese)