树蝴蝶多糖的免疫调节活性研究（英文）
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摘要
本文采用超声波辅助水提、乙醇沉淀、DEAE-52纤维素阴离子交换层析法分离纯化得到从树蝴蝶原料中提取的中性多糖LKY-I。以不同浓度的多糖为原料,研究了LKY-I的体外免疫调节活性。结果表明,LKY-I在125~500μg/mL浓度范围内对RAW 264.7细胞无明显毒性。LKY-I可显著增强巨噬细胞吞噬中性红的能力,当多糖浓度为500μg/mL时,吸光度值可达到1.39,显著高于阳性对照组LPS。LKY-I可促进巨噬细胞RAW 264.7分泌NO,最大分泌量可达55.42±1.79μM;与正常对照组相比,LKY-I促进巨噬细胞分泌IL-6、TNF-α细胞因子,分泌量分别增加了102.3%和289.5%。在125~500μg/mL浓度范围内,LKY-I可显著提高i NOS、IL-6及TNF-α基因的表达。LKY-I协同LPS和Con A能显著促进小鼠脾淋巴细胞的增殖。研究结果表明,LKY-I多糖具有良好的体外免疫调节活性,可以作为一种有益健康的成分应用到食品工业中。
The aim of this study was to determine the immunomodulatory activities of the neutral polysaccharide LKY-I extracted fromLobaria kurokawae Yoshim.The polysaccharide LKY-I was isolated sequentially using ultrasonic-assisted water extraction,ethanol precipitation,diethylaminoethyl cellulose(DEAE-52)ion-exchange chromatography,and gel permeation chromatography.We investigated the immunomodulatory activities using different concentrations of LKY-I.The polysaccharide LKY-I at a concentration range of 31.25–500μg/mL had no obvious in vitro toxic effects in macrophages(RAW 264.7).The phagocytic ability of macrophages treated with 500μg/mL LKY-I was higher than that of the macrophages in the positive control group treated with lipopolysaccharide(LPS);absorbance,1.39.In addition,LKY-I induced the production of nitric oxide(NO)up to 55.42±1.79μM.Compared with the control group,the LKY-I group showed an increase in the secretion of tumor necrosis factorα(TNF-α)and interleukin 6(IL-6)by 102.3%and 289.5%,respectively.Furthermore,LKY-I at a dose of125–500μg/mL showed a marked increase in the mR NA expression levels of inducible nitric oxide synthase(iN OS),TNF-α,and interleukin 6(IL-6).LKY-I in combination with LPS or concanavalin A(ConA)significantly promoted the proliferation of lymphocytes.Thus,our findings suggest that LKY-I has immunomodulatory effects and can be used as a health-promoting food supplement.
The aim of this study was to determine the immunomodulatory activities of the neutral polysaccharide LKY-I extracted fromLobaria kurokawae Yoshim.The polysaccharide LKY-I was isolated sequentially using ultrasonic-assisted water extraction,ethanol precipitation,diethylaminoethyl cellulose(DEAE-52)ion-exchange chromatography,and gel permeation chromatography.We investigated the immunomodulatory activities using different concentrations of LKY-I.The polysaccharide LKY-I at a concentration range of 31.25–500μg/mL had no obvious in vitro toxic effects in macrophages(RAW 264.7).The phagocytic ability of macrophages treated with 500μg/mL LKY-I was higher than that of the macrophages in the positive control group treated with lipopolysaccharide(LPS);absorbance,1.39.In addition,LKY-I induced the production of nitric oxide(NO)up to 55.42±1.79μM.Compared with the control group,the LKY-I group showed an increase in the secretion of tumor necrosis factorα(TNF-α)and interleukin 6(IL-6)by 102.3%and 289.5%,respectively.Furthermore,LKY-I at a dose of125–500μg/mL showed a marked increase in the mR NA expression levels of inducible nitric oxide synthase(iN OS),TNF-α,and interleukin 6(IL-6).LKY-I in combination with LPS or concanavalin A(ConA)significantly promoted the proliferation of lymphocytes.Thus,our findings suggest that LKY-I has immunomodulatory effects and can be used as a health-promoting food supplement.
引文
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[10]Wang M,Yang X B,Zhao J W,et al.Structural characterization and macrophage immunomodulatory activity of a novel polysaccharide from Smilax glabra Roxb[J].Carbohydrate Polymers,2016,156:390-402
[11]Liu W,Wang H,Yu J,et al.Structure,chain conformation,and immunomodulatory activity of the polysaccharide purified from Bacillus Calmette Guerin formulation[J].Carbohydrate Polymers,2016,150:149-158
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[13]Jia Q,Sang M K.Characterization and immunomodulatory activities of polysaccharides extracted from green alga Chlorella ellipsoidea[J].International Journal of Biological Macromolecules,2017,95:106-114
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[15]Gordon S.Phagocytosis:An Immunobiologic Process[J].Immunity,2016,44(3):463
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[18]Gordon S.Phagocytosis:An Immunobiologic Process[J].Immunity,2016,44(3):463-475
[19]Schepetkin I A,Quinn M T.Botanical polysaccharides:Macrophage immunomodulation and therapeutic potential[J].International Immunopharmacology,2006,6(3):317-333
[20]Zhang X,Li Y,Tai G,et al.Effects of activin a on the activities of the mouse peritoneal macrophages[J].Chinese Journal of Immunology,2005,2(1):63-67
[21]Drozina G,Kohoutek J,Nishiya T,et al.Sequential modifications in class II transactivator isoform 1 induced by lipopolysaccharide stimulate major histocompatibility complex class II transcription in macrophages[J].Journal of Biological Chemistry,2006,281(52):39963-39970
[22]Chen X,Lu J,Zhang Y,et al.Studies of macrophage immuno-modulating activity of polysaccharides isolated from Paecilomyces tenuipes[J].International Journal of Biological Macromolecules,2008,43(3):252-256
[23]Bogdan C.Nitric oxide synthase in innate and adaptive immunity:an update[J].Trends in Immunology,2015,36(3):161-178
[24]Antonelli A,Ferri C,Ferrari S M,et al.Serum levels of proinflammatory cytokines interleukin-1beta,interleukin-6,and tumor necrosis factor alpha in mixed cryoglobulinemia[J].Arthritis&Rheumatology,2009,60(12):3841-3847
[25]Yoon S B,Lee Y J,Park S K,et al.Anti-inflammatory effects of Scutellaria baicalensis water extract on LPS-activated RAW264.7 macrophages[J].Journal of Ethnopharmacology,2009,125(2):286-290
[26]Schepetkin I A,Quinn M T.Botanical polysaccharides:Macrophage immunomodulation and therapeutic potential[J].International Immunopharmacology,2006,6(3):317-333
[27]Nakamura T,Suzuki H,Wada Y,et al.Fucoidan induces nitric oxide production via p38 mitogen-activated protein kinase and NF-kappa B-dependent signaling pathways through macrophage scavenger receptors[J].Biochemical&Biophysical Research Communications,2006,343(1):286-294
[28]Mebius R E,Kraal G.Structure and function of the spleen[J].Nature Reviews Immunology,2005,5(8):606-616
[29]Sarangi I,Ghosh D,Bhutia S K,et al.Anti-tumor and immunomodulating effects of Pleurotus ostreatus mycelia-derived proteoglycans[J].International Immunopharmacology,2006,6(8):1287-1297
[2]Pei J,Wang Z,Ma H,et al.Structural features and antitumor activity of a novel polysaccharide from alkaline extract of Phellinus linteus mycelia[J].Carbohydrate Polymers,2015,115:472-477
[3]Kiho T,Morimoto H,Kobayashi T,et al.Effect of a polysaccharide(TAP)from the fruiting bodies of Tremella aurantia on glucose metabolism in mouse liver[J].Bioscience Biotechnology&Biochemistry,2000,64(2):417-419
[4]Liu Y,Zhang B,Ibrahim S A,et al.Purification,characterization and antioxidant activity of polysaccharides from Flammulina velutipes residue[J].Carbohydrate Polymers,2016,145:71-77
[5]Zhang M,Cui S W,Cheung P C K,et al.Antitumor polysaccharides from mushrooms:a review on their isolation process,structural characteristics and antitumor activity[J].Trends in Food Science&Technology,2007,18(1):4-19
[6]Zong A,Cao H,Wang F.Anticancer polysaccharides from natural resources:A review of recent research[J].Carbohydrate Polymers,2012,90(4):1395-1410
[7]Xin Y F,You Z Q,Gao H Y,et al.Protective effect of Lycium barbarum polysaccharides against doxorubicin-induced testicular toxicity in rats[J].Phytotherapy Research,2012,26(5):716-721
[8]Shen C Q,Chen J.Structural analysis and antioxidant and antitumor activity of polysaccharide LKY-I from Lobaria koorkauae Yoshim[J].Food Science,2017,21:119-125
[9]Wu D T,Meng L Z,Wang L Y,et al.Chain conformation and immunomodulatory activity of a hyperbranched polysaccharide from Cordyceps sinensis[J].Carbohydrate Polymers,2014,110C(38):405-414
[10]Wang M,Yang X B,Zhao J W,et al.Structural characterization and macrophage immunomodulatory activity of a novel polysaccharide from Smilax glabra Roxb[J].Carbohydrate Polymers,2016,156:390-402
[11]Liu W,Wang H,Yu J,et al.Structure,chain conformation,and immunomodulatory activity of the polysaccharide purified from Bacillus Calmette Guerin formulation[J].Carbohydrate Polymers,2016,150:149-158
[12]Xu D,Wang H,Wei Z,et al.Charaterization and immunomodulatory activities of polysaccharide isolated from Pleurotus eryngii[J].International Journal of Biological Macromolecules,2016,92:30-36
[13]Jia Q,Sang M K.Characterization and immunomodulatory activities of polysaccharides extracted from green alga Chlorella ellipsoidea[J].International Journal of Biological Macromolecules,2017,95:106-114
[14]Mosser D M,Edwards J P.Mosser DM,Edwards JPExploring the full spectrum of macrophage activation.Nat Rev Immunol[J].Nature Reviews Immunology,2009,8(12):958-969
[15]Gordon S.Phagocytosis:An Immunobiologic Process[J].Immunity,2016,44(3):463
[16]Hao N B,LüM H,Fan Y H,et al.Macrophages in tumor microenvironments and the progression of tumors[J].Clinical&Developmental Immunology,2012,2012(11):948098
[17]Mosser D M,Edwards J P.Exploring the full spectrum of macrophage activation[J].Nature Reviews Immunology,2008,8(12):958-969
[18]Gordon S.Phagocytosis:An Immunobiologic Process[J].Immunity,2016,44(3):463-475
[19]Schepetkin I A,Quinn M T.Botanical polysaccharides:Macrophage immunomodulation and therapeutic potential[J].International Immunopharmacology,2006,6(3):317-333
[20]Zhang X,Li Y,Tai G,et al.Effects of activin a on the activities of the mouse peritoneal macrophages[J].Chinese Journal of Immunology,2005,2(1):63-67
[21]Drozina G,Kohoutek J,Nishiya T,et al.Sequential modifications in class II transactivator isoform 1 induced by lipopolysaccharide stimulate major histocompatibility complex class II transcription in macrophages[J].Journal of Biological Chemistry,2006,281(52):39963-39970
[22]Chen X,Lu J,Zhang Y,et al.Studies of macrophage immuno-modulating activity of polysaccharides isolated from Paecilomyces tenuipes[J].International Journal of Biological Macromolecules,2008,43(3):252-256
[23]Bogdan C.Nitric oxide synthase in innate and adaptive immunity:an update[J].Trends in Immunology,2015,36(3):161-178
[24]Antonelli A,Ferri C,Ferrari S M,et al.Serum levels of proinflammatory cytokines interleukin-1beta,interleukin-6,and tumor necrosis factor alpha in mixed cryoglobulinemia[J].Arthritis&Rheumatology,2009,60(12):3841-3847
[25]Yoon S B,Lee Y J,Park S K,et al.Anti-inflammatory effects of Scutellaria baicalensis water extract on LPS-activated RAW264.7 macrophages[J].Journal of Ethnopharmacology,2009,125(2):286-290
[26]Schepetkin I A,Quinn M T.Botanical polysaccharides:Macrophage immunomodulation and therapeutic potential[J].International Immunopharmacology,2006,6(3):317-333
[27]Nakamura T,Suzuki H,Wada Y,et al.Fucoidan induces nitric oxide production via p38 mitogen-activated protein kinase and NF-kappa B-dependent signaling pathways through macrophage scavenger receptors[J].Biochemical&Biophysical Research Communications,2006,343(1):286-294
[28]Mebius R E,Kraal G.Structure and function of the spleen[J].Nature Reviews Immunology,2005,5(8):606-616
[29]Sarangi I,Ghosh D,Bhutia S K,et al.Anti-tumor and immunomodulating effects of Pleurotus ostreatus mycelia-derived proteoglycans[J].International Immunopharmacology,2006,6(8):1287-1297