Jiang Yiping, Qi Xiaohui, Gao Kai, Liu Wenjun, Li Na, Cheng Ningbo, Ding Gang, Huang Wenzhe, Wang Zhenzhong, Xiao Wei
State Key Laboratory of New-tech for Chinese Medicine Pharmaceutical Processes, Jiangsu Kanion Pharmaceutical Co. Ltd., 58 Haichangnan Road, Lianyungang, Jiangsu Province, People's Republic of China.
Jinan Health and Medical Technology Co., Ltd., Jinan high-tech zone picked Chinese road, 2766, Jinan, Shandong Province, People's Republic of China.
Glycoconj J. 2016 Oct;33(5):755-61. doi: 10.1007/s10719-016-9669-z. Epub 2016 Apr 30.
Four Astragalus polysaccharides (APS1-APS4) were isolated from the water extract of Radix Astragali and purified through ethanol precipitation with 20 %, 40 %, 60 % and 80 % ethanol, respectively. The total sugar content was measured by sulfuric acid-phenol method. Their molecular weight was determined using high performance gel permeation chromatography (HPGPC) and their monosaccharide composition was analyzed by reversed-phase high performance liquid chromatography (HPLC) after pre-column derivatization. Then the immunobiologic activity of APS was evaluated by the experiment of spleen lymphocytes proliferation in vitro. The data suggested that precipitation by different concentration of ethanol will obtain different molecular weight APS, the higher concentration of ethanol the smaller molecular weight for APS. The molecular weights of four APS were 257.7 kDa, 40.1 kDa, 15.3 kDa and 3.2 kDa. Monosaccharide composition analysis indicated that APS1 consisted of glucose only, and APS2 all consisted of arabinose. APS3 consisted of rhamnose, glucose, galactose and arabinose and APS4 consisted of galactose and arabinose, in a molar ratio of 1:10.76:6.55:12 and 3.02:1. The result of immunobiologic activity assay showed that both APS2 and APS3 can effectively stimulate normal spleen lymphocyte proliferation in vitro. Apart from this, the effect of APS2 also showed dose dependent tendency from 6.25 μg/mL to 800 μg/mL. The result of this research indicated that Astragalus polysaccharides, which consist of arabinose and their molecular weight between 15.2 kDa to 40.1 kDa, neither too high nor too low, had significant immune activity.
从黄芪水提取物中分离出四种黄芪多糖(APS1 - APS4),分别用20%、40%、60%和80%乙醇沉淀法进行纯化。采用硫酸 - 苯酚法测定总糖含量。使用高效凝胶渗透色谱法(HPGPC)测定其分子量,并通过柱前衍生化后的反相高效液相色谱法(HPLC)分析其单糖组成。然后通过体外脾淋巴细胞增殖实验评估APS的免疫生物学活性。数据表明,不同浓度乙醇沉淀可得到不同分子量的APS,乙醇浓度越高,APS分子量越小。四种APS的分子量分别为257.7 kDa、40.1 kDa、15.3 kDa和3.2 kDa。单糖组成分析表明,APS1仅由葡萄糖组成,APS2全部由阿拉伯糖组成。APS3由鼠李糖、葡萄糖、半乳糖和阿拉伯糖组成,APS4由半乳糖和阿拉伯糖组成,摩尔比分别为1:10.76:6.55:12和3.02:1。免疫生物学活性测定结果表明,APS2和APS3均可有效刺激体外正常脾淋巴细胞增殖。除此之外,APS2在6.25 μg/mL至800 μg/mL范围内其作用也呈剂量依赖性趋势。本研究结果表明,由阿拉伯糖组成且分子量在15.2 kDa至40.1 kDa之间、既不太高也不太低的黄芪多糖具有显著的免疫活性。