铁皮石斛中性多糖的分离纯化及抗炎作用

Isolation and Purification of Polysaccharide and Anti-inflammatory Effect of Dendrobium officinale Neutral Polysaccharide

目的:从铁皮石斛中分离纯化得到铁皮石斛多糖,对其进行结构表征,并探究其抗炎作用。方法:以铁皮石斛为原料,通过水提醇沉法得到铁皮石斛粗多糖,再经过DEAE-52纤维素离子交换树脂和葡聚糖凝胶Sephadex G-100从粗多糖中分离纯化得到多糖组分DOP-1。利用紫外扫描法、气相色谱法、高效凝胶色谱、红外光谱和核磁共振扫描等技术对多糖DOP-1进行结构分析,并测定DOP-1对小鼠腹腔巨噬细胞RAW 264.7细胞增殖抑制效果,及其对LPS诱导的RAW264.7细胞NO、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和白介素-1β(interleukin-1β,IL-1β)表达的影响。结果:分离纯化得到的多糖DOP-1为鼠李糖、甘露糖、葡萄糖和半乳糖组成的均一性多糖,单糖摩尔比为0.6:9.0:2.7:2.9,分子量为59.2 kDa。DOP-1对RAW 264.7细胞增殖无显著性抑制,与模型组相比,DOP-1能显著抑制NO、TNF-α和IL-1β的表达量。结论:本实验得到的铁皮石斛多糖结构明确,且具有较好的抗炎活性,提取工艺条件具有可行性。本研究可为铁皮石斛多糖在抗炎功效产品的开发利用提供理论依据。

Objective:To isolate and purify polysaccharides from Dendrobium officinale,characterize their structure and explore their anti-inflammatory effects.Methods:Polysaccharides from Dendrobium officinale were obtained by water extraction and alcohol precipitation.DOP-1 was purified by DEAE-52 cellulose ion exchange resin and Sephadex G-100.Ultraviolet scanning,GC,high-performance gel chromatography,infrared spectroscopy and nuclear magnetic resonance scanning were used to analyze the structure of DOP-1 polysaccharide,and the inhibitory effect of DOP-1 on the proliferation of RAW 264.7 cells was determined.And its effect on the expression of NO,TNF-αand IL-1βin RAW264.7 cells induced by LPS.Results:DOP-1 was a homogeneous polysaccharide composed of rhamnose,mannose,glucose and galactose with a monosaccharide molar ratio of 0.6:9.0:2.7:2.9 and a molecular weight of 59.2 kDa.DOP-1 did not significantly inhibit the proliferation of RAW 264.7 cells.Compared with the model group,DOP-1 significantly inhibited the expression of NO,TNF-αand IL-1β.Conclusions:The polysaccharides of Dendrobium officinaleobtained in this study had clear structure and good anti-inflammatory activity,and the extraction conditions were feasible.This study would provide theoretical basis for the development and utilization of polysaccharides of Dendrobium officinale with anti-inflammatory effect.