金针菇子实体多糖FVPB2对小鼠T淋巴细胞和巨噬细胞的免疫调节作用
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摘要
从金针菇子实体中分离纯化得到均一多糖FVPB2,其分子量为15kDa,是由葡萄糖、半乳糖、岩藻糖和甘露糖组成的吡喃型杂多糖,利用C57BL/6小鼠脾淋巴细胞和骨髓巨噬细胞研究FVPB2对免疫功能的影响,体外免疫实验表明,FVPB2能促进T淋巴细胞激活并分泌肿瘤坏死因子和干扰素γ细胞因子,同时还能够促进巨噬细胞产生一氧化氮,分泌白介素-1β、白介素-6和肿瘤坏死因子-α细胞因子。本研究以首次从金针菇子实体中获得均一多糖FVPB2为研究对象,观察其对T细胞和巨噬细胞的免疫调节活性,研究结果表明其具有良好的免疫调节活性和潜在的生物学功能。
The homogeneous polysaccharides FVPB2 from the fruiting body of Flammulina filiformis were separated and purified for the first time.The purified polysaccharide was a heteropolysaccharide consisting of D-glucose,D-galactose,L-fucose,and D-mannose,with an average molecular weight of about 1.5×10~4Da.The immunoactivity of FVPB2 in vitro was evaluated on spleen lymphocyte and bone marrow macrophage in C57BL/6 mice.In vitro immunomodulatory studies showed FVPB2 induced T lymphocyte activation,and stimulated TNF and IFN-γsecretion of T lymphocyte.FVPB2 also could induce NO production and IL-1β,IL-6and TNF-αsecretion of bone marrow macrophage.It was proved that the homogenous polysaccharide FVPB2could enhance non-specific immune response and specific immune responses in vitro.
The homogeneous polysaccharides FVPB2 from the fruiting body of Flammulina filiformis were separated and purified for the first time.The purified polysaccharide was a heteropolysaccharide consisting of D-glucose,D-galactose,L-fucose,and D-mannose,with an average molecular weight of about 1.5×10~4Da.The immunoactivity of FVPB2 in vitro was evaluated on spleen lymphocyte and bone marrow macrophage in C57BL/6 mice.In vitro immunomodulatory studies showed FVPB2 induced T lymphocyte activation,and stimulated TNF and IFN-γsecretion of T lymphocyte.FVPB2 also could induce NO production and IL-1β,IL-6and TNF-αsecretion of bone marrow macrophage.It was proved that the homogenous polysaccharide FVPB2could enhance non-specific immune response and specific immune responses in vitro.
引文
Chang HL,Lei SL,Yu CL,Zhu ZG,Chen NN,Wu SG,2009.Effect of Flammulina velutipes polysaccharides on production of cytokines by murine immunocytes and serum levels of cytokines in tumor-bearing mice.Journal of Chinese Medicinal Materials,32(4):561-563
Chen ZY,Yan MX,Xiang BK,Zhu SD,Huang XM,2001.Effect of Flammulina velutipes polysaccharides on splenic lymphocyte transformation rate,natural killer cell and interleukin-2 production in mice.Chinese Journal of Integrated Traditional and Western Medicine,1:105-107(in Chinese)
Dai YC,Yang ZL,2018.Notes on the nomenclature of five important edible fungi in China.Mycosystema,37(12):1572-1577(in Chinese)
Feng T,2016.Stuies on isolation,purification,identifying structure and immune activity of polysaccharides from Flammulina velutipes fruiting body.Master Thesis,Jilin Agricultural University,Changchun.67-78(in Chinese)
Feng T,Jia W,Wang WH,Lin CH,Fan H,Zhang JS,Bao HY,2016.Structural characterization and immunological activities of a novel water-soluble polysaccharide from the fruiting bodies of culinary-medicinal winter mushroom,Flammulina velutipes(Agaricomycetes).International Journal of Medicinal Mushrooms,18(9):807-819
Feng T,Wang WH,Zhang JS,Fan H,Jia W,Bao HY,2016.Effects of Flammulina velutipes different extracts on tumor cells.Mycosystema,35(10):1234-1243(in Chinese)
Fu YX,2012.Research on extraction,purification and cell immune activity of polysaccharides from the base of Flammulina velutipes stipe.Master Thesis,Nanjing Agricultural University,Nanjing.56-73(in Chinese)
He YL,Ying Y,Xu YL,Su JF,Luo H,Wang HF,2005.Effects of Lycium barbarum polysaccharide on tumor microenvironment T-lymphocyte subsets and dendritic cells in H22-bearing mice.Journal of Chinese Integrative Medicine,3(5):374-377
Jia W,Feng J,Zhang JS,Lin CC,Wang WH,Zhang JS,Chen HG,2017.Structural characteristics of the novel polysaccharide FVPA1 from winter culinary-medicinal mushroom Flammulina velutipes(Agaricomycetes),capable of enhancing natural killer cell activity against K562 tumor cells.International Journal of Medicinal Mushrooms,19(6):535
Jose ML,Castro R,Arranz JA,Jesus L,2007.Immunomodulating activities of acidic sulphated polysaccharides obtained from the seaweed Ulva rigida C.Agardh.International Immunopharmacology,7(7):879-888
Leung MY,Fung KP,Choy YM,1997.The isolation and characterization of an immunomodulatory and anti-tumor polysaccharide preparation from Flammulina velutipes.Immunopharmacology,35(3):255-263
Li B,Bao YQ,2017.Progress of CD19-specific modified chimeric antigen receptor T cells in the treatment of Bcell lymphoma and leukemia.Journal of Clinical Hematology,5:82-85(in Chinese)
Li SM,Ren Q,Li M,Tian JH,2009.Extraction of polysaccharide in Flammulina velutipes and evaluation of its function in cosmetics.Acta Edulis Fungi,31:72-73(in Chinese)
Liu CX,2016.Precision pharmacy:investigating new drug research and development from translational medicine to precision medicine.Drug Evaluation Research,39(1):1-18(in Chinese)
Ma DD,Wang WH,Jia W,Zhang JS,Ge XY,2017.Application of edible fungus polysaccharide as additives in farming practices.Acta Agriculture Shanghai,33(3):166-170(in Chinese)
Methven A,Hughes KW,Petersen RH,2000.Flammulina RFLP patterns identify species and show biogeographical patterns within species.Mycologia,92:1064
Ming W,Xia L,Xu XY,2014.Antioxidant and immunomodulatory activities of a polysaccharide from Flammulina velutipes.Journal of Traditional Chinese Medicine,34(6):733-740
Nauta AJ,Fibbe WE,2007.Immunomodulatory properties of mesenchymal stromal cells.Blood,110(10):3499-3506
Otagiri K,Ohkuma T,Ikekawa T,Tanka S,1983.Intensification of antitumor-immunity by protein-bound polysaccharide,EA6,derived from Flammulina velutipes(Curt.ex Fr.)Sing.combined with murine leukemia L1210 vaccine in animal experiments.Journal of Pharmacobio-dynamics,6(2):96
Tang C,Hoo PC,Tan LT,Pusparajah P,Khan TM,Lee LH,Goh BH,Chan KG,2016.Golden needle mushroom:a culinary medicine with evidenced-based biological activities and health promoting properties.Frontiers in Pharmacology,7:474
Tang QJ,Zhang JS,Pan YJ,Martin ZK,Werner R,Fan H,2005.Activation of normal and tumor-bearing mouse macrophages by active polysaccharides from Ganoderma lucidum.Current Immunology,25(1):49-52(in Chinese)
Wang MZ,Chen ZZ,2011.Progress on studies and applications of modern combined techniques in biomacromolecule drugs.Food and Drug,13(3):209-212(in Chinese)
Wang PM,Liu XB,Dai YC,Horak E,Steffen K,Yang ZL,2018.Phylogeny and species delimitation of Flammulina:taxonomic status of winter mushroom in East Asia and a new European species identified using an integrated approach.Mycological Progress,17:1013-1030
Wang WH,Jia AQ,Zhang JS,Yang Y,Zhang H,Sun WB,Jia W,2017.Anti-newcastle disease virus activity of polysaccharide fractions from Flammulina velutipes fruiting bodies and their effect on chicken immune organ indices.Acta Edulis Fungi,4:78-84(in Chinese)
Wang WH,Zhang JS,Feng T,Deng J,Lin CC,Fan H,Yu WJ,Bao HY,Jia W,2018.Structural elucidation of a polysaccharide from Flammulina velutipes and its immunomodulation activities on mouse B lymphocytes.Scientific Reports,8:3120
Wang YF,Wang X,Yin HP,2008.Purification,characterization and immune activity of polysaccharides from Flammulina velutipes mycelium.Chinese Journal of Natural Medicines,6(4):312-315(in Chinese)
Wen MJ,Yu MH,Yu X,2018.Discovery of cancer therapy by inhibition of negative immune regulation-the 2018Nobel Prize in Physiology or Medicine.Journal of Capital Medical University,39:765-769(in Chinese)
Yan YS,Yao WB,Zhang J,Zhou S,Zhang X,Gao XD,2011.Structural investigation and activation of macrophages in vitro of a novel polysaccharide from Flammulina velutipes mycelium.Journal of China Pharmaceutical University,42(2):164-168(in Chinese)
Yang Y,Liu CC,Jia W,Liu YF,Yang Y,Zhang JS,Zhou CY,2008.Study on ultrafiltration separation and immunocompetence of polysaccharides from Grifola frondosa.Food Science,29(9):277-280(in Chinese)
Yang YP,2018.The current situation of China’s edible fungus industrial production from the perspective of enterprises in 2017.Edible and Medicinal Mushrooms,26(1):13-14(in Chinese)
Yoshioka Y,Sano T,Ikekawa T,1973.Studies on antitumor polysaccharides of Flammulina velutipes(Curt.ex Fr.)Sing.I.Chemical&Pharmaceutical Bulletin,21:1772-1776
Yu Q,2014.Immunomodulatary effect and underlying mechanism of Ganoderma atrum polysaccharide.PhDDissertation,Nanchang University,Nanchang.70-78(in Chinese)
Zhang JS,Tang QJ,Zhou C,Jia W,Da SL,Nguyen LD,Reutter W,Fan H,2010.GLIS,a bioactive proteoglycan fraction from Ganoderma lucidum,displays anti-tumour activity by increasing both humoral and cellular immune response.Life Sciences,87(19):628-637
Zhu YY,Wang R,Wei W,Zhu XY,Wang YQ,Wang Q,2015.Anti-tumor and immunomodulatory effects of exopolysaccharides from Flammulina rossica on H22tumor-bearing mice.Mycosystema,34(4):772-778(in Chinese)
Zipfel PF,2009.Complement and immune defense:from innate immunity to human diseases.Immunology Letters,126:1-7
陈芝芸,严茂祥,项柏康,朱曙东,黄小民,2001.金针菇多糖对小鼠体内脾淋巴细胞转化、自然杀伤细胞活性及白细胞介素-2产生的影响.中国中西医结合杂志,1:105-107
戴玉成,杨祝良,2018.中国五种重要食用菌学名新注.菌物学报,37(12):1572-1577
冯婷,2016.金针菇子实体多糖的分离纯化结构鉴定及免疫活性研究.吉林农业大学硕士论文,长春.67-78
冯婷,汪文翰,张劲松,樊华,贾薇,包海鹰,2016.金针菇不同萃取物对肿瘤细胞的影响.菌物学报,35(10):1234-1243
傅亚欣,2012.金针菇菌柄基部多糖的提取、纯化及细胞免疫活性研究.南京农业大学硕士论文,南京.56-73
李斌,鲍扬漪,2017.CD19-CART技术在B细胞淋巴瘤及白血病中的应用进展.临床血液学杂志,5:82-85
李守勉,任清,李明,田景花,2009.金针菇多糖的提取及其美容功效评价.食用菌学报,31(5):72-73
刘昌孝,2016.精准药学:从转化医学到精准医学探讨新药发展.药物评价研究,39(1):1-18
马丹丹,汪雯翰,贾薇,张劲松,葛欣芸,2017.上海农学报,33(3):166-170
唐庆九,张劲松,潘迎捷,Martin Zimmerman-Kordmann,Werner Reutter,樊华,2005.灵芝活性多糖GLIS对正常和荷瘤小鼠骨髓巨噬细胞的激活作用.现代免疫学,25(1):49-52
王铭章,陈执中,2011.近代联用技术在生物大分子药物研究中应用进展.食品与药品,13(3):209-212
汪雯翰,贾爱卿,张劲松,杨焱,张赫,孙文博,贾薇,2017.金针菇子实体多糖抗新城疫病毒活性及对鸡免疫器官的影响.食用菌学报,4:78-84
王玉峰,王旻,尹鸿萍,2008.金针菇菌丝体中多糖的分离、结构鉴定及免疫学活性.中国天然药物,6(4):312-315
温铭杰,于明航,王玺,2018.癌症免疫治疗的利器-2018年诺贝尔生理学或医学奖简介.首都医科大学学报,39:765-769
闫昳姝,姚文兵,张健,邹珊,张先,高向东,2011.富锗金针菇多糖FVP1结构的初步鉴定及其对巨噬细胞功能调节作用.中国药科大学学报,42(2):164-168
杨阳,刘承初,贾薇,刘艳芳,杨炎,张劲松,周昌艳,2008.灰树花多糖的超滤分离及免疫活性研究.食品科学,29(9):277-280
杨勇萍,2018.从企业角度看2017年我国食用菌工厂化生产的现状.食药用菌,1:13-14
余强,2014.黑灵芝多糖的免疫调节活性及其作用机制.南昌大学博士论文,南昌.70-78
朱宴妍,王瑞,魏巍,朱相杨,王永强,王琦,2015.柳生金针菇胞外粗多糖对小鼠肝癌H22移植性肿瘤的抑制作用和免疫功能调节的研究.菌物学报,34(4):772-778
Chen ZY,Yan MX,Xiang BK,Zhu SD,Huang XM,2001.Effect of Flammulina velutipes polysaccharides on splenic lymphocyte transformation rate,natural killer cell and interleukin-2 production in mice.Chinese Journal of Integrated Traditional and Western Medicine,1:105-107(in Chinese)
Dai YC,Yang ZL,2018.Notes on the nomenclature of five important edible fungi in China.Mycosystema,37(12):1572-1577(in Chinese)
Feng T,2016.Stuies on isolation,purification,identifying structure and immune activity of polysaccharides from Flammulina velutipes fruiting body.Master Thesis,Jilin Agricultural University,Changchun.67-78(in Chinese)
Feng T,Jia W,Wang WH,Lin CH,Fan H,Zhang JS,Bao HY,2016.Structural characterization and immunological activities of a novel water-soluble polysaccharide from the fruiting bodies of culinary-medicinal winter mushroom,Flammulina velutipes(Agaricomycetes).International Journal of Medicinal Mushrooms,18(9):807-819
Feng T,Wang WH,Zhang JS,Fan H,Jia W,Bao HY,2016.Effects of Flammulina velutipes different extracts on tumor cells.Mycosystema,35(10):1234-1243(in Chinese)
Fu YX,2012.Research on extraction,purification and cell immune activity of polysaccharides from the base of Flammulina velutipes stipe.Master Thesis,Nanjing Agricultural University,Nanjing.56-73(in Chinese)
He YL,Ying Y,Xu YL,Su JF,Luo H,Wang HF,2005.Effects of Lycium barbarum polysaccharide on tumor microenvironment T-lymphocyte subsets and dendritic cells in H22-bearing mice.Journal of Chinese Integrative Medicine,3(5):374-377
Jia W,Feng J,Zhang JS,Lin CC,Wang WH,Zhang JS,Chen HG,2017.Structural characteristics of the novel polysaccharide FVPA1 from winter culinary-medicinal mushroom Flammulina velutipes(Agaricomycetes),capable of enhancing natural killer cell activity against K562 tumor cells.International Journal of Medicinal Mushrooms,19(6):535
Jose ML,Castro R,Arranz JA,Jesus L,2007.Immunomodulating activities of acidic sulphated polysaccharides obtained from the seaweed Ulva rigida C.Agardh.International Immunopharmacology,7(7):879-888
Leung MY,Fung KP,Choy YM,1997.The isolation and characterization of an immunomodulatory and anti-tumor polysaccharide preparation from Flammulina velutipes.Immunopharmacology,35(3):255-263
Li B,Bao YQ,2017.Progress of CD19-specific modified chimeric antigen receptor T cells in the treatment of Bcell lymphoma and leukemia.Journal of Clinical Hematology,5:82-85(in Chinese)
Li SM,Ren Q,Li M,Tian JH,2009.Extraction of polysaccharide in Flammulina velutipes and evaluation of its function in cosmetics.Acta Edulis Fungi,31:72-73(in Chinese)
Liu CX,2016.Precision pharmacy:investigating new drug research and development from translational medicine to precision medicine.Drug Evaluation Research,39(1):1-18(in Chinese)
Ma DD,Wang WH,Jia W,Zhang JS,Ge XY,2017.Application of edible fungus polysaccharide as additives in farming practices.Acta Agriculture Shanghai,33(3):166-170(in Chinese)
Methven A,Hughes KW,Petersen RH,2000.Flammulina RFLP patterns identify species and show biogeographical patterns within species.Mycologia,92:1064
Ming W,Xia L,Xu XY,2014.Antioxidant and immunomodulatory activities of a polysaccharide from Flammulina velutipes.Journal of Traditional Chinese Medicine,34(6):733-740
Nauta AJ,Fibbe WE,2007.Immunomodulatory properties of mesenchymal stromal cells.Blood,110(10):3499-3506
Otagiri K,Ohkuma T,Ikekawa T,Tanka S,1983.Intensification of antitumor-immunity by protein-bound polysaccharide,EA6,derived from Flammulina velutipes(Curt.ex Fr.)Sing.combined with murine leukemia L1210 vaccine in animal experiments.Journal of Pharmacobio-dynamics,6(2):96
Tang C,Hoo PC,Tan LT,Pusparajah P,Khan TM,Lee LH,Goh BH,Chan KG,2016.Golden needle mushroom:a culinary medicine with evidenced-based biological activities and health promoting properties.Frontiers in Pharmacology,7:474
Tang QJ,Zhang JS,Pan YJ,Martin ZK,Werner R,Fan H,2005.Activation of normal and tumor-bearing mouse macrophages by active polysaccharides from Ganoderma lucidum.Current Immunology,25(1):49-52(in Chinese)
Wang MZ,Chen ZZ,2011.Progress on studies and applications of modern combined techniques in biomacromolecule drugs.Food and Drug,13(3):209-212(in Chinese)
Wang PM,Liu XB,Dai YC,Horak E,Steffen K,Yang ZL,2018.Phylogeny and species delimitation of Flammulina:taxonomic status of winter mushroom in East Asia and a new European species identified using an integrated approach.Mycological Progress,17:1013-1030
Wang WH,Jia AQ,Zhang JS,Yang Y,Zhang H,Sun WB,Jia W,2017.Anti-newcastle disease virus activity of polysaccharide fractions from Flammulina velutipes fruiting bodies and their effect on chicken immune organ indices.Acta Edulis Fungi,4:78-84(in Chinese)
Wang WH,Zhang JS,Feng T,Deng J,Lin CC,Fan H,Yu WJ,Bao HY,Jia W,2018.Structural elucidation of a polysaccharide from Flammulina velutipes and its immunomodulation activities on mouse B lymphocytes.Scientific Reports,8:3120
Wang YF,Wang X,Yin HP,2008.Purification,characterization and immune activity of polysaccharides from Flammulina velutipes mycelium.Chinese Journal of Natural Medicines,6(4):312-315(in Chinese)
Wen MJ,Yu MH,Yu X,2018.Discovery of cancer therapy by inhibition of negative immune regulation-the 2018Nobel Prize in Physiology or Medicine.Journal of Capital Medical University,39:765-769(in Chinese)
Yan YS,Yao WB,Zhang J,Zhou S,Zhang X,Gao XD,2011.Structural investigation and activation of macrophages in vitro of a novel polysaccharide from Flammulina velutipes mycelium.Journal of China Pharmaceutical University,42(2):164-168(in Chinese)
Yang Y,Liu CC,Jia W,Liu YF,Yang Y,Zhang JS,Zhou CY,2008.Study on ultrafiltration separation and immunocompetence of polysaccharides from Grifola frondosa.Food Science,29(9):277-280(in Chinese)
Yang YP,2018.The current situation of China’s edible fungus industrial production from the perspective of enterprises in 2017.Edible and Medicinal Mushrooms,26(1):13-14(in Chinese)
Yoshioka Y,Sano T,Ikekawa T,1973.Studies on antitumor polysaccharides of Flammulina velutipes(Curt.ex Fr.)Sing.I.Chemical&Pharmaceutical Bulletin,21:1772-1776
Yu Q,2014.Immunomodulatary effect and underlying mechanism of Ganoderma atrum polysaccharide.PhDDissertation,Nanchang University,Nanchang.70-78(in Chinese)
Zhang JS,Tang QJ,Zhou C,Jia W,Da SL,Nguyen LD,Reutter W,Fan H,2010.GLIS,a bioactive proteoglycan fraction from Ganoderma lucidum,displays anti-tumour activity by increasing both humoral and cellular immune response.Life Sciences,87(19):628-637
Zhu YY,Wang R,Wei W,Zhu XY,Wang YQ,Wang Q,2015.Anti-tumor and immunomodulatory effects of exopolysaccharides from Flammulina rossica on H22tumor-bearing mice.Mycosystema,34(4):772-778(in Chinese)
Zipfel PF,2009.Complement and immune defense:from innate immunity to human diseases.Immunology Letters,126:1-7
陈芝芸,严茂祥,项柏康,朱曙东,黄小民,2001.金针菇多糖对小鼠体内脾淋巴细胞转化、自然杀伤细胞活性及白细胞介素-2产生的影响.中国中西医结合杂志,1:105-107
戴玉成,杨祝良,2018.中国五种重要食用菌学名新注.菌物学报,37(12):1572-1577
冯婷,2016.金针菇子实体多糖的分离纯化结构鉴定及免疫活性研究.吉林农业大学硕士论文,长春.67-78
冯婷,汪文翰,张劲松,樊华,贾薇,包海鹰,2016.金针菇不同萃取物对肿瘤细胞的影响.菌物学报,35(10):1234-1243
傅亚欣,2012.金针菇菌柄基部多糖的提取、纯化及细胞免疫活性研究.南京农业大学硕士论文,南京.56-73
李斌,鲍扬漪,2017.CD19-CART技术在B细胞淋巴瘤及白血病中的应用进展.临床血液学杂志,5:82-85
李守勉,任清,李明,田景花,2009.金针菇多糖的提取及其美容功效评价.食用菌学报,31(5):72-73
刘昌孝,2016.精准药学:从转化医学到精准医学探讨新药发展.药物评价研究,39(1):1-18
马丹丹,汪雯翰,贾薇,张劲松,葛欣芸,2017.上海农学报,33(3):166-170
唐庆九,张劲松,潘迎捷,Martin Zimmerman-Kordmann,Werner Reutter,樊华,2005.灵芝活性多糖GLIS对正常和荷瘤小鼠骨髓巨噬细胞的激活作用.现代免疫学,25(1):49-52
王铭章,陈执中,2011.近代联用技术在生物大分子药物研究中应用进展.食品与药品,13(3):209-212
汪雯翰,贾爱卿,张劲松,杨焱,张赫,孙文博,贾薇,2017.金针菇子实体多糖抗新城疫病毒活性及对鸡免疫器官的影响.食用菌学报,4:78-84
王玉峰,王旻,尹鸿萍,2008.金针菇菌丝体中多糖的分离、结构鉴定及免疫学活性.中国天然药物,6(4):312-315
温铭杰,于明航,王玺,2018.癌症免疫治疗的利器-2018年诺贝尔生理学或医学奖简介.首都医科大学学报,39:765-769
闫昳姝,姚文兵,张健,邹珊,张先,高向东,2011.富锗金针菇多糖FVP1结构的初步鉴定及其对巨噬细胞功能调节作用.中国药科大学学报,42(2):164-168
杨阳,刘承初,贾薇,刘艳芳,杨炎,张劲松,周昌艳,2008.灰树花多糖的超滤分离及免疫活性研究.食品科学,29(9):277-280
杨勇萍,2018.从企业角度看2017年我国食用菌工厂化生产的现状.食药用菌,1:13-14
余强,2014.黑灵芝多糖的免疫调节活性及其作用机制.南昌大学博士论文,南昌.70-78
朱宴妍,王瑞,魏巍,朱相杨,王永强,王琦,2015.柳生金针菇胞外粗多糖对小鼠肝癌H22移植性肿瘤的抑制作用和免疫功能调节的研究.菌物学报,34(4):772-778