| 摘要: |
| 【目的】从灵芝孢子粉中提取纯化多糖，研究灵芝多糖对小鼠细胞免疫功能的影响，为灵芝开发利用提供试验依据。【方法】利用水煮醇沉法从灵芝孢子粉中提取粗多糖，经反复冻融、Sevag法脱蛋白和Sepharose CL-6B柱层析，纯化得灵芝多糖(GLP)。通过HPLC鉴定GLP的纯度，G.C.分析其单糖组成。采用MTT法、中性红试验、Griess法、比色法和ELISA法，分别检测GLP对小鼠脾淋巴细胞增殖、腹腔巨噬细胞吞噬功能、NO产量和iNOS活性以及TNF-α产量的影响。【结果】GLP为白色粉末，分子质量为51 ku，HPLC分析GLP为单一狭窄对称峰，其单糖组成为葡萄糖(Glc)和半乳糖(Gal)，二者物质的量之比为16.8∶1。GLP能够刺激小鼠T淋巴细胞增殖，并呈现剂量依赖关系；当GLP质量浓度达到100 μg/mL时，能显著增强腹腔巨噬细胞的吞噬能力，提高其分泌iNOS的活力和NO的生产量；同时，当GLP质量浓度≥200 μg/mL时，能显著促进腹腔巨噬细胞分泌TNF-α的能力，具有剂量依赖性。【结论】GLP能够增强小鼠免疫细胞活性，有望成为重要的生物免疫调节剂。 |
| 关键词: 灵芝多糖 脾淋巴细胞 腹腔巨噬细胞 免疫调节 |
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| 基金项目:黑龙江省自然科学基金项目(C201227)；大庆师范学院博士启动基金项目(11NR16) |
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| Extraction and purification of polysaccharide from Ganoderma lucidum and its immunological activities |
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ZHANG Li-xia,ZHANG Ya-jun,ZHANG Li-ping
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| Abstract: |
| 【Objective】This study focused on extraction and purification of polysaccharide from Ganoderma lucidum spore powder and its effects on cell immunity of mice,to improve the development and utilization of Ganoderma lucidum.【Method】Water boiling and ethanol method was used to extract polysaccharide from Ganoderma lucidum spore powder,and Ganoderma lucidum polysaccharide (GLP) was obtained after repeated freezing and thawing,Sevag protein removal,Sepharose CL-6B column chromatography,and purification.The purity was identified by HPLC,and its monosaccharide composition was analyzed by GC.Effects of Ganoderma lucidum polysaccharides on spleen lymphocyte proliferation,phagocytosis of peritoneal macrophages,NO production,iNOS activity and TNF-α yield were detected by MTT,neutral red test,Griess method,colorimetric method,and ELISA method,respectively.【Result】Obtained GLP was white powder with molecular weight of 51 ku.HPLC analysis showed it had a single narrow symmetrical peak.GLP consisted of glucose (Glc) and galactose (Gal) with a molar ratio of 16.8∶1.GLP stimulated the proliferation of T lymphocyte in mice with dose dependent relationship.When the concentration of GLP reached 100 μg/mL,the phagocytic activity of peritoneal macrophage was significantly enhanced,and iNOS production activity of peritoneal macrophage and NO production were improved.When GLP mass concentration was larger than 200 μg/mL,TNF-α secretion ability of macrophage was significantly promoted with dose dependent relationship.【Conclusion】Ganoderma lucidum polysaccharide could enhance the activity of immune cells in mice,and act as an important biological immune regulator. |
| Key words: Ganoderma lucidum polysaccharide lymphocyte macrophage immune-regulation |
