艾根多糖的结构特征及免疫活性研究

Structural characteristics and immunomodulatory activity of polysaccharide from the root of Artemisia argyi

目的:艾根为种植艾叶(Artemisiae Argyi Folium)药材的副产品,富含黄酮和多糖,在民间常用作滋补食材,但其药用和保健价值未受重视。方法:该研究利用水提醇沉法提取分离得到艾根多糖(AARP),采用高效凝胶渗透色谱、PMP-HPLC和红外光谱初步解析其结构。CCK-8法检测AARP对脾淋巴细胞增殖的影响;中性红吞噬实验测定RAW264.7细胞吞噬能力,ELISA测定一氧化氮(NO)、肿瘤坏死因子(TNF-α)、白细胞介素(IL)-6及IL-1β的水平;采用环磷酰胺免疫抑制小鼠模型评价其体内免疫活性;最后通过DPPH自由基(DPPH·)、羟自由基(·OH)、超氧阴离子(O_(2)·^(-))自由基清除能力及ABTS、FRAP总抗氧化能力实验评价其抗氧化活性。结果:结果表明AARP主成分分子量为5.9 kDa,由葡萄糖、半乳糖、阿拉伯糖、甘露糖、鼠李糖、半乳糖醛酸、葡萄糖醛酸(摩尔比62.91∶17.17∶8.55∶5.75∶1.96∶1.72∶1.94)和微量的木糖组成。该多糖在10~1000μg·mL^(-1)浓度范围内能显著促进脾淋巴细胞的增殖,提高巨噬细胞的吞噬能力及NO、TNF-α、IL-6、IL-1β的释放;并可明显逆转环磷酰胺所致小鼠体重减轻,恢复器官指数和碳廓清能力;且对·OH、O_(2)·^(-)、DPPH和ABTS自由基有不同程度的清除能力。结论:该研究揭示了艾根多糖的基本结构并证实其具有优异的免疫调节及抗氧化活性,可为艾蒿植物资源利用和功能性产品的开发提供理论参考。

OBJECTIVE The root of Artemisia argyi is rich inflavonoids and polysaccharides,which is used as a tonic in the folk but it’s medicinal and health value has not been valued.METHODS In this study the polysaccharide(AARP)in it was extracted and separated by water extraction and alcohol precipitation.Its monosaccharide composition and molar ratio were detected by PMP-HPLC,and the structure was preliminarily resolved by FT-IR.The effects of AAPR on the proliferation of spleen lymphocytes cells were detected by CCK-8 method.The level of nitric oxide(NO),tumor necrosis factor(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)released by cells was measured by EILISA.Neutral red phagocytosis experiments and carbon clearance experiment were used to evaluate the phagocytic capacity.1-diphenyl-2-trinitrophenylhydrazine(DPPH)free radical,superoxide anion free radical(O_(2)·^(-)),hydroxyl free radical(·OH)scavenging ability and total antioxidant capacity(ABTS,FRAP)test were carried out to investigate its antioxidant activity.RESULTS The results showed AARP was composed of glucose,galactose,arabinose,mannose,rhamnose,galacturonic acid,glucuronic acid(molar ratio 62.91∶17.17∶8.55∶5.75∶1.96∶1.72∶1.94)with a molecular weight of 5.9 kDa,which could significantly promote the proliferation of spleen lymphocytes,the phagocytic ability of macrophages and the release of NO,TNF-α,IL-6 and IL-1β.AARP showed the·OH,DPPH and O_(2)·^(-)scavenging ability of by EC_(50) of 1.61,2.94 and 6.32 mg·mL-1.CONCLUSION This study revealed the structure of polysaccharide from A.argyi root with excellent immune regulationand antioxidant activities.