枝瑚菌多糖的结构及其免疫调节活性
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摘要
本研究以枝瑚菌中分离纯化的枝瑚菌多糖(RF-S)为研究对象,运用红外光谱技术研究了其结构;运用细胞学技术,从体外水平,以巨噬细胞和淋巴细胞为靶细胞,研究了RF-S的免疫调节活性。结果表明:RF-S多糖结构明显,无其他结构,为多糖纯品,且其多糖中的单糖是以吡喃糖苷的形式存在。随着RF-S质量浓度的增加,极显著促进巨噬细胞、T淋巴细胞和B淋巴细胞的增殖(p <0.01),并呈现一定的剂量依赖关系。同时,当RF-S质量浓度为5μg/m L时,能极显著地促进巨噬细胞吞噬中性红和荧光微球(p <0.01),并能极显著地刺激巨噬细胞分泌IL-2、IL-10、TNFα和释放NO(p <0.01);同时对刺激巨噬细胞分泌IL-6有显著效果(p <0.05)。综上所述,RF-S具有良好的免疫调节活性。
In this study,polysaccharides(RF-S) was isolated from the Ramaria flaccida(Fr.) Quél,which was used as the research object of study.Its structure was studied by infra-red spectrum,and the immunomodulatory activity of RF-S was studied in vitro by using cytological techniques and using macrophages and lymphocytes as target cells.The results showed that the polysaccharide structure of RF-S was typical without any other structure,and it was a kind of pure polysaccharide.And the monosaccharide in its polysaccharides existed in the form of pyranose.In addition,with the increase of RF-S concentration,the proliferation of macrophages,T lymphocytes and B lymphocytes could be extremely significantly promoted(p < 0.01),and showed a dose-dependent relationship.Meanwhile,when the mass concentration of RF-S was 5 μg/m L,it could extremely significantly promote phagocytosis of neutral red and fluorescent microspheres in macrophage,and could extremely significantly stimulate macrophages to secrete IL-2,IL-10,TNF alpha and release NO(p < 0.01).It also had a significant effect on stimulating the secretion of IL-6 in macrophages(p < 0.05).In conclusion,polysaccharides from the Ramaria flaccida(Fr.)Quél had good immunomodulatory activity.
In this study,polysaccharides(RF-S) was isolated from the Ramaria flaccida(Fr.) Quél,which was used as the research object of study.Its structure was studied by infra-red spectrum,and the immunomodulatory activity of RF-S was studied in vitro by using cytological techniques and using macrophages and lymphocytes as target cells.The results showed that the polysaccharide structure of RF-S was typical without any other structure,and it was a kind of pure polysaccharide.And the monosaccharide in its polysaccharides existed in the form of pyranose.In addition,with the increase of RF-S concentration,the proliferation of macrophages,T lymphocytes and B lymphocytes could be extremely significantly promoted(p < 0.01),and showed a dose-dependent relationship.Meanwhile,when the mass concentration of RF-S was 5 μg/m L,it could extremely significantly promote phagocytosis of neutral red and fluorescent microspheres in macrophage,and could extremely significantly stimulate macrophages to secrete IL-2,IL-10,TNF alpha and release NO(p < 0.01).It also had a significant effect on stimulating the secretion of IL-6 in macrophages(p < 0.05).In conclusion,polysaccharides from the Ramaria flaccida(Fr.)Quél had good immunomodulatory activity.
引文
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[22]顾菲菲,李佳,杨晨东,等.碱提灵芝多糖的分离纯化、结构表征及免疫活性评价[J].中草药,2018,49(10):2359-2364.
[23]谭庆,罗炳德,张培,等.青蒿琥酯对高温内毒素复合因素下巨噬细胞吞噬功能的影响[J].中国工业医学杂志,2005,18(2):98-100.
[24]华慧,张波,李向阳,等.华支睾吸虫脂肪酸结合蛋白Cs FABP对巨噬细胞免疫功能的影响[J].中国病原生物学杂志,2017,12(11):1058-1061.
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[26]宋广磊,王奎武,张虹,等.毛轴蕨多糖的分离纯化、单糖组成及对巨噬细胞RAW264.7免疫活性的影响[J].中国药学杂志,2017,52(11):931-935.
[27]张晓音,张珊珊,吴旻,等.β-胡萝卜素对脂多糖刺激巨噬细胞RAW264.7炎症因子的影响及其机制[J].中国免疫学杂志,2017,33(6):838-843.
[28]姜雨薇,崔逢德,李红花,等.桦褐孔菌多糖对脂多糖刺激小鼠巨噬细胞促炎性细胞因子表达的影响[J].延边大学医学学报,2015,38(3):174-177.
[29]徐红霞,蒋媛媛,洪雅雯,等.野生枝瑚菌多糖的提取及单糖组成分析[J].食品工业科技,2016,37(21):255-259
[30]赵鹤鹏.玉米须多糖的制备及其生物活性研究[D].吉林:吉林化工学院,2017.
[31]张莘莘.黑灵芝多糖的抗肿瘤活性及其分子机制初探[D].南昌:南昌大学,2014.
[32]李华,窦晓兰,陆启玉.葡萄状枝瑚菌粗多糖的提取及其清除DPPH自由基活性[J].食用菌学报,2012,19(3):69-72.
[2]罗晓莉,李建英,张沙沙,等.云南三种特色野生食用菌营养成分分析与评价[J].食品工业,2017,38(5):277-280.
[3]何强,陈文强,解修超,等.陕西佛坪野生“刷把菌”营养成分的分析与评价[J].西北农林科技大学学报:自然科学版,2018(12):1-6.
[4]庞海月.泡囊侧耳细胞毒活性成分的分离鉴定及野生药用珊瑚菌液体优化培养[D].福州:福建师范大学,2015.
[5]苏凤贤,杨全文,李彩霞,等.密枝瑚菌多糖提取工艺参数优化[J].天然产物研究与开发,2011,23(4):751-754.
[6]鹿洪亮.灵芝多糖有效成分提取及纯化方法综述[J].科技信息,2009,27:429-430.
[7]魏华,张娣,陆玲,等.几种食用菌多糖的提取与抗氧化性研究[J].南京师大学报:自然科学版,2017,40(2):72-75.
[8]Faccin-Galhardi L C,Yamamoto K A,Ray S,et al.The in vitro antiviral property of Azadirachta indica polysaccharides for poliovirus[J].Ethnopharmacol,2012,142(1):86-90.
[9]Tang H L,Chen C,Wang S K,et al.Biochemical analysis and hypoglycemic activity of a polysaccharide isolated from the fruit of Lycium barbarum L[J].Int J Biol Macromol,2015,77:235-242.
[10]Li S P,Zhang G H,Zeng Q,et al.Hypoglycemic activity of polysaccharide,with antioxidation,isolated from cultured Cordyceps mycelia[J].Phytomedicine,2006,13(6):428-433.
[11]李志涛,赵娟娟,王敏,等.复合食用菌多糖的免疫活性研究[J].食品研究与开发,2017,38(5):39-42.
[12]徐红霞.野生枝瑚菌多糖的提取、分离纯化和抗氧化性研究[D].杭州:浙江大学,2017.
[13]李美华,孙玉侠,曾阳,等.粗柄羊肚菌菌丝体多糖及胞外多糖的提取工艺[J].菌物研究,2017,15(3):195-200.
[14]白岚.真菌多糖的研究进展---真菌多糖的结构、提纯和应用[J].河北林果研究,2009,24(4):445-448.
[15]丛大鹏.南极海茸(Durvillaea antarctica)多糖提取、衍生物制备及免疫活性研究[D].青岛:中国海洋大学,2015.
[16]盛家荣,曾令辉,翟春,等.多糖的提取、分离及结构分析[J].广西师范学院学报:自然科学版,1999,16(4):49-54.
[17]赵美峰,魏超妮,剡建华.硫酸-苯酚法测定黑果枸杞中多糖的含量[J].中医药信息,2015,32(2):58-59.
[18]孙波,鞠萍,杨科,等.枸杞鲜果冻干制品中枸杞多糖的含量及抗氧化活性研究[J].食品科技,2014,39(10):223-226.
[19]唐颖,安利佳.FTIR法分析铵盐胁迫对蓝莓组培苗的影响[J].北方园艺,2018(4):48-53.
[20]Ni W,Zhang X,Wang B,et al.Antitumor activities and immunomodulatory effects of ginseng neutral polysaccharides in combination with 5-fluorouracil[J].Journal of medicinal food,2010,13(2):270-277.
[21]Han H,Chen Y,Bi H,et al.In vivo antimalarial activity of ginseng extracts[J].Pharmaceutical Biology,2011,49(3):283-289.
[22]顾菲菲,李佳,杨晨东,等.碱提灵芝多糖的分离纯化、结构表征及免疫活性评价[J].中草药,2018,49(10):2359-2364.
[23]谭庆,罗炳德,张培,等.青蒿琥酯对高温内毒素复合因素下巨噬细胞吞噬功能的影响[J].中国工业医学杂志,2005,18(2):98-100.
[24]华慧,张波,李向阳,等.华支睾吸虫脂肪酸结合蛋白Cs FABP对巨噬细胞免疫功能的影响[J].中国病原生物学杂志,2017,12(11):1058-1061.
[25]Noula S,Nicole S H,Daniel J L,et al.Essential role for TAX1 BP1 in the termination of TNF-α,IL-1 and LPS mediated NF and JNK signaling[J].EMBO J,2007,26(17):3910-3922.
[26]宋广磊,王奎武,张虹,等.毛轴蕨多糖的分离纯化、单糖组成及对巨噬细胞RAW264.7免疫活性的影响[J].中国药学杂志,2017,52(11):931-935.
[27]张晓音,张珊珊,吴旻,等.β-胡萝卜素对脂多糖刺激巨噬细胞RAW264.7炎症因子的影响及其机制[J].中国免疫学杂志,2017,33(6):838-843.
[28]姜雨薇,崔逢德,李红花,等.桦褐孔菌多糖对脂多糖刺激小鼠巨噬细胞促炎性细胞因子表达的影响[J].延边大学医学学报,2015,38(3):174-177.
[29]徐红霞,蒋媛媛,洪雅雯,等.野生枝瑚菌多糖的提取及单糖组成分析[J].食品工业科技,2016,37(21):255-259
[30]赵鹤鹏.玉米须多糖的制备及其生物活性研究[D].吉林:吉林化工学院,2017.
[31]张莘莘.黑灵芝多糖的抗肿瘤活性及其分子机制初探[D].南昌:南昌大学,2014.
[32]李华,窦晓兰,陆启玉.葡萄状枝瑚菌粗多糖的提取及其清除DPPH自由基活性[J].食用菌学报,2012,19(3):69-72.