细虫草胞外多糖对小鼠腹腔巨噬细胞免疫功能研究
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摘要
本实验在体外条件下,以人工发酵培养的细虫草胞外多糖OgE、OgE-F1和OgE-F2作用于小鼠腹腔巨噬细胞RAW264.7,通过测定其对巨噬细胞的增殖率、代谢MTT活力、NO分泌和吞噬能力的影响,评价细虫草胞外多糖的免疫调节活性。结果表明,细虫草多糖对巨噬细胞无细胞毒性,且能促进巨噬细胞代谢MTT活力;在0.2 mg/mL～1.0 mg/mL浓度范围内,多糖呈剂量依赖性的促进巨噬细胞分泌NO水平和吞噬能力。本研究表明,细虫草多糖能有效地增强小鼠巨噬细胞的活性,潜在地可改善小鼠的先天性免疫调节。
In order to investigate effects of exopolysaccharides from Ophiocordceps gracilis on the mouse peritoneal macrophage in vitro, different concentrations of OgE, OgE-F1 and OgE-F2 were cocultured with the mouse peritoneal macrophage. The metabolic activity and the proliferative activity of macrophages were measured by cell counting kit-8(CCK-8) and MTT assay, respectively. The nitric oxide was detected by Griess reagent. The phagocytic activity of peritoneal macrophages was measured by neutral red method. The exopolysaccharide from O. gracilis had no cytotoxic activity on peritoneal macrophage and could enhance the metabolic activity of peritoneal macrophage. The production of NO was increased and the phagocytic activity of peritoneal macrophage was improved in a dose-depended manner when macrophages were exposed to exopolysaccharides of O. gracilis ranging from 0.2 mg/mL to 1.0 mg/mL. The experiments demonstrated that the exopolysaccharides from O. gracilis could effectively enhance the immunity of mouse peritoneal macrophages and improve the innate immune regulation of mice.
In order to investigate effects of exopolysaccharides from Ophiocordceps gracilis on the mouse peritoneal macrophage in vitro, different concentrations of OgE, OgE-F1 and OgE-F2 were cocultured with the mouse peritoneal macrophage. The metabolic activity and the proliferative activity of macrophages were measured by cell counting kit-8(CCK-8) and MTT assay, respectively. The nitric oxide was detected by Griess reagent. The phagocytic activity of peritoneal macrophages was measured by neutral red method. The exopolysaccharide from O. gracilis had no cytotoxic activity on peritoneal macrophage and could enhance the metabolic activity of peritoneal macrophage. The production of NO was increased and the phagocytic activity of peritoneal macrophage was improved in a dose-depended manner when macrophages were exposed to exopolysaccharides of O. gracilis ranging from 0.2 mg/mL to 1.0 mg/mL. The experiments demonstrated that the exopolysaccharides from O. gracilis could effectively enhance the immunity of mouse peritoneal macrophages and improve the innate immune regulation of mice.
引文
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[14] 田维毅,王文佳,李海峰,等.中性红法检测巨噬细胞吞噬功能的实验条件的优化[J].贵阳中医学院学报,2009,31(2):23-26.
[15] 苏进娟.九州虫草多糖的提取纯化、理化性质及免疫调节、抗氧化活性研究[D].山东:山东大学,2014:1-70.
[16] LEE J S,KWON D S,LEE K R,et al.Mechanism of macrophage activation induced by polysaccharide from Cordyceps militaris culture broth[J].Carbohyd Polym,2015,120:29-37.
[17] BRüNE B.Nitric oxide:NO apoptosis or turning it ON?[J].Cell Death Differ,2003,10 (8):864-869.
[18] 王永敏,祝文兴,安利国,等.蛹虫草胞外多糖的提取纯化及免疫活性研究[J].食品与药品,2009,11(3):8-11.
[19] 肖建辉,梁宗琦,胡锡阶,等.古尼虫草多糖及其解聚物的免疫活性[J].免疫学杂志,2005,21(1):51-53.
[2] 索菲娅,苏俊,姜彦成,等.新疆虫草与冬虫夏草中甘露醇、多糖和氨基酸的含量比较研究[J].新疆农业科学,2008,45(3):517-521.
[3] 索菲娅,苏俊,姜彦成,等.新疆虫草与冬虫夏草中腺苷和虫草素的HPLC法测定比较[J].中国民族医药杂志,2008,3(3):65-67.
[4] 陈露,安利国.虫草多糖的免疫调节作用及其抗肿瘤活性的研究[J].山东师范大学学报(自然科学版),2009,24(4):109-112.
[5] LEE J S,KWON J S,WON D P,et al.Study on macrophage activation and structural characteristics of purified polysaccharide from the liquid culture broth of Cordyceps militaris[J].Carbohyd Polym,2010,82 (3):982-988.
[6] WANG L Y,CHEONG K L,WU D T,et al.Fermentation optimization for the production of bioactive polysaccharides from Cordyceps sinensis fungus UM01[J].Int J Biol Macromol,2015,79:180-185.
[7] 许周善,周晓燕.冬虫夏草多糖的研究进展[J].工业微生物,2000,30(1):56-57.
[8] LERIO J M,CASTRO R,ARRANZ J A,et al.Immunomodulating activities of acidic sulphated polysaccharides obtained from the seaweed Ulva rigida C.Agardh[J].Int Immunopharmacol,2007,7 (7):879-888.
[9] LEE K Y,JEON Y J.Macrophage activation by polysaccharide isolated from Astragalus membranaceus[J].Int Immunopharmacol,2005,5(7-8):1225-1233.
[10] 杨静,丁重阳,顾正华,等.以豆浆为培养基的三种药食用真菌发酵过程多糖变化的研究[J].工业微生物,2015,45(5):17-22.
[11] 童微,余强,李虎,等.铁皮石斛多糖化学修饰及其对免疫活性的影响[J].食品科学,2017,38(7):155-160.
[12] LIU Z G,XING J,HUANG Y,et al.Activation effect of Ganoderma lucidum polysaccharides liposomes on murine peritoneal macrophages[J].In J Biol Macromol,2015,82:973-978.
[13] 杭宜岭,于婧,林祥娜,等.具有改善去卵巢小鼠钙磷吸收作用的益生菌筛选与鉴定[J].工业微生物,2018,48(2):12-17.
[14] 田维毅,王文佳,李海峰,等.中性红法检测巨噬细胞吞噬功能的实验条件的优化[J].贵阳中医学院学报,2009,31(2):23-26.
[15] 苏进娟.九州虫草多糖的提取纯化、理化性质及免疫调节、抗氧化活性研究[D].山东:山东大学,2014:1-70.
[16] LEE J S,KWON D S,LEE K R,et al.Mechanism of macrophage activation induced by polysaccharide from Cordyceps militaris culture broth[J].Carbohyd Polym,2015,120:29-37.
[17] BRüNE B.Nitric oxide:NO apoptosis or turning it ON?[J].Cell Death Differ,2003,10 (8):864-869.
[18] 王永敏,祝文兴,安利国,等.蛹虫草胞外多糖的提取纯化及免疫活性研究[J].食品与药品,2009,11(3):8-11.
[19] 肖建辉,梁宗琦,胡锡阶,等.古尼虫草多糖及其解聚物的免疫活性[J].免疫学杂志,2005,21(1):51-53.