铁皮石斛多糖对脂多糖诱导的巨噬细胞免疫调节作用

黄杰, 章莹, 严婧, 肖榕, 夏伯候, 吴萍, 林丽美, 廖端芳

中国药学杂志 ›› 2017, Vol. 52 ›› Issue (7) : 548-552.

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中国药学杂志 ›› 2017, Vol. 52 ›› Issue (7) : 548-552. DOI: 10.11669/cpj.2017.07.007
药食两用中药专栏

铁皮石斛多糖对脂多糖诱导的巨噬细胞免疫调节作用

  • 黄杰1,2 , 章莹1,2 , 严婧1,2, 肖榕1,2 , 夏伯候1,2*, 吴萍1,2*, 林丽美1,2 , 廖端芳1,2
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Immunomodulatory Effect of Dendrobium officinale Polysaccharide on Macrophage RAW264.7 Induced by LPS

  • HUANG Jie1,2, ZHANG Ying1,2, YAN Jing1,2, XIAO Rong1,2, XIA Bo-hou1,2* , WU Ping1,2*, LIN Li-mei1,2, LIAO Duan-fang1,2
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摘要

目的 研究经分段醇沉所得铁皮石斛多糖(DOP)对巨噬细胞(RAW264.7)的免疫调节作用。方法 采用分步醇沉法得到不同相对分子质量的铁皮石斛多糖,四唑盐比色法(MTT)检测DOP对脂多糖(LPS)诱导巨噬细胞的保护作用,中性红法检测巨噬细胞的吞噬活性,酶联免疫吸附实验法(Elisa)检测培养液中肿瘤坏死因子α(TNF-α)、白介素6(IL-6)、转化生长因子-β1(TGF-β1)的含量,格里斯试剂比色法(Griess)检测一氧化氮(NO)的释放水平,流式细胞仪检测细胞的极化分型。结果 醇浓度为75%时,醇沉的多糖得率最大;DOP能促进RAW264.7细胞增殖,增强巨噬细胞的吞噬活性,降低巨噬细胞TNF-α、IL-6、NO的释放水平,促进细胞分泌TGF-β1并向M2型极化,其效应具有显著性差异(P<0.01)。结论 铁皮石斛多糖能够对RAW264.7细胞起到免疫调节的作用,其机制可能与其促进细胞的吞噬活性、降低炎症因子释放水平和改变细胞极化分型有关。

Abstract

OBJECTIVE To study the immunoregulatory effect of Dendrobium officinale polysaccharide (DOP) obtained by step ethanol precipitation on macrophages and explore its action mechanisms. METHODS Step ethanol precipitation method was used to get DOPs of different molecular weights, MTT method was used to detect the protective effect of DOP on macrophages induced by LPS, neutral red method was used to detect the phagocytosis activity of macrophages, ELISA method was used to detect the contents of TNF-α, IL-6, and TGF-β1, and Griess method was used to detect the NO release quantity. RESULTS The yield of DOP was the largest when the ethanol concentration was 75%. DOP could promote the proliferation and phagocytosis activity of RAW264.7 cells, reduce the release of TNF-α, IL-6 and NO, and promote the secretion of TGF-β1 and the proportion of M2 significantly (P<0.01). CONCLUSION Dendrobium officinale polysaccharide has immune regulation effect on RAW264.7 cells, and the mechanism may be related to the promotion of cell phagocytosis activity, inhibition of inflammatory factor release, promotion of TGF-β1 secretion, and increase of the proportion of M2.

关键词

铁皮石斛多糖 / 分段醇沉 / 巨噬细胞 / 免疫调节

Key words

Dendrobium officinale polysaccharide / step ethanol precipitation / macrophage / immune regulation

引用本文

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黄杰, 章莹, 严婧, 肖榕, 夏伯候, 吴萍, 林丽美, 廖端芳. 铁皮石斛多糖对脂多糖诱导的巨噬细胞免疫调节作用[J]. 中国药学杂志, 2017, 52(7): 548-552 https://doi.org/10.11669/cpj.2017.07.007
HUANG Jie, ZHANG Ying, YAN Jing, XIAO Rong, XIA Bo-hou, WU Ping, LIN Li-mei, LIAO Duan-fang. Immunomodulatory Effect of Dendrobium officinale Polysaccharide on Macrophage RAW264.7 Induced by LPS[J]. Chinese Pharmaceutical Journal, 2017, 52(7): 548-552 https://doi.org/10.11669/cpj.2017.07.007
中图分类号: R282   

参考文献

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基金

湖南省教育厅重点项目(14A109)资助项目;湖南省发改委项目([2013]1199)资助项目;湖南省科技厅科研条件创新专项(2012TT2012)资助项目;湖南省研究生省级科研创新项目(CX2015B336)资助项目
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