铁皮石斛中性多糖分离纯化及其体内免疫调节作用研究  

Seperation and Purification of Neutral Polysaccharides from Dendrobium officinale and Study of Its Immunoregulatory Effects

作　　者：雷思敏[1,2,3] 肖榕 章莹[2,3] 黄杰 严婧 林丽美 夏伯候 吴萍[1,2,3] LEI Simin;XIAO Rong;ZHANG Ying;HUANG Jie;YAN Jing;LIN Limei;XIA Bohou;WU Ping(Key Laboratory of Pharmaceutical Sciences, School of Pharmacy, Hunan University of Chinese Medicine,Changsha 410208 Hunan,China;Hunan Research Center of Engineering Technology for Rapid Test and Removal of Toxic and Harmful Substances in Chinese Medicine,Changsha 410208 Hunan,China;Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province,Changsha 410208 Hunan,China)

机构地区：[1]湖南中医药大学药学院,湖南长沙410208 [2]湖南省中药不良成分快速检测及脱除工程技术研究中心/中药有毒物质防控技术湖南省工程实验室,湖南长沙410208 [3]湘产大宗药材品质评价湖南省重点实验室,湖南长沙410208

出　　处：《中药新药与临床药理》2018年第6期748-753,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：湖南省教育厅重点项目(14A109)

摘　　要：目的分离鉴定铁皮石斛中的中性多糖,探讨铁皮石斛中性多糖(DOP-1)对环磷酰胺所致免疫功能低下小鼠免疫功能的影响。方法水提醇沉法提取铁皮石斛粗多糖、酶-Sevage法脱蛋白;DEAE-纤维素52离子交换层析法分离纯化粗多糖;红外扫描和柱前衍生化HPLC法分析多糖组分及构型;选用BALB/c雄性小鼠,采用环磷酰胺建立免疫抑制小鼠动物模型,实验设正常组,模型组,左旋咪唑阳性组,DOP-1高、低剂量组,铁皮石斛全茎高、低剂量组。观察比较各小鼠脾脏和胸腺指数、血清中白细胞介素-6(IL-6)、白细胞介素-10(IL-10)和干扰素-γ(IFN-γ)含量及血清溶血素水平。结果铁皮石斛粗多糖经柱层析分离纯化后得到中性多糖(DOP-1),DOP-1主要由甘露糖、鼠李糖、葡萄糖、半乳糖、木糖和阿拉伯糖组成,其摩尔比分别为3.8∶0.25∶1.5∶3.8∶2.1∶1.3。动物实验表明:与模型组比较,DOP-1及全茎可显著增加胸腺指数,显著下调血清中IL-6(P<0.05)和IL-10(P<0.05)的含量,DOP-1调节作用优于全茎和阳性组;DOP-1高剂量组的IFN-γ水平也有显著性降低(P<0.01),并能增强免疫抑制小鼠血清溶血素水平(P<0.01)。结论铁皮石斛中性多糖可缓解环磷酰胺对小鼠的免疫抑制作用。Objective To isolate and identify the neutral polysaccharides from Dendrobium officinale and observe the effects of Dendrobium officinale polysaccharides-1(DOP-1) on immune functions in immunosuppressed mice induced by cyclophosphamide. Methods The crude Dendrobium officinale polysaccharides were preliminarily extracted with water,precipitated in ethyl alcohol,proteins were removed by enzyme-Savage method and then the polysaccharides were purified by DEAE-52 ion exchange chromatography. Monosaccharide compositions in neutral polysaccharides were separated and detected by infrared scanning and pre-column derivatization- HPLC. The male BALB/c mice that selected to develop immune-suppressed mouse models induced by cyclophos-phamide (CP) were assigned to eight groups:normal group,model group,Levamisole positive group,DOP-1 high-dose group,DOP-1 low-dose group, whole stem high- dose group, whole stem low-dose group. All the groups were given corresponding dosage of drugs with intragastric administration to observe the spleen index and thymus index,the contents of IL-6,IL-10 and IFN-γ in serum,the level of serum hemolysin of mice and then these indices were compared. Results DOP-1 were isolated and purified by column chromatography from crude polysaccharides. DOP-1 was mainly composed of mannose,rhamnose,glucose,galactose,xylose and arabinose,and their molar ratio is 3.8∶0.25∶1.5∶3.8∶2.1∶1.3. Trials on mice showed that compared with the model group,DOP-1 groups and the whole stem group can strongly increase the spleen and thymus indexes,decrease the concentrations of IL-6(P<0.05) and IL-10 (P<0.01) in serum. The regulation function of DOP-1 is superior to whole stem (high- dose) and positive control group. Moreover,the IFN-γ level of DOP-1 high-dose group is lower than model group (P<0.01),and the level of serum hemolysin of immunosuppressed mice is improved (P<0.01). Conclusion DOP-1 can alleviate CP-induced immunosuppressive effects in mice.

关 键 词：铁皮石斛 中性多糖 环磷酰胺 免疫调节 

分 类 号：R285.5[医药卫生—中药学]