乌梅丸水提液中多糖理化性质分析及预防结肠炎的作用

目的:考察乌梅丸水提液中多糖含量,建立HPLC法分析乌梅丸多糖分子量及单糖组成方法,观察其对右旋葡聚糖硫酸钠(DSS)所致小鼠溃疡性结肠炎(UC)的预防作用,为乌梅丸临床用药及药理活性成分的研究提供依据。方法:采用水提醇沉法提取多糖;硫酸-苯酚法测定乌梅丸多糖含量;凝胶色谱法-龙智达分子量工作站测定多糖分子量;PMP柱前衍生化法HPLC分析单糖组成;DSS(3%)法建立UC小鼠模型。ICR小鼠50只随机分为5组:正常组,模型组,乌梅丸多糖2.5%,5%,10%剂量组,每组10只。造模前7天,在乌梅丸多糖治疗各组的小鼠饮水中添加乌梅丸多糖,一直给药维持至实验结束(第14天)。结果:葡萄糖在0~0.08 mg·m L-1浓度范围内呈良好的线性关系(r=0.998);右旋葡聚糖酐标准品分子量在2500~2000000 Da范围内呈良好线性关系(r=0.998);乌梅丸水提液中多糖含量为40.3%,纯化的乌梅丸多糖糖含量为91.6%;分子量范围在171343~525009 Da之间,分布系数D=1.18;其多糖的单糖组成主要为甘露糖、葡萄糖醛酸、葡萄糖、半乳糖和木糖组成,其相对含量比为1.8:1.0:19.3:32.8:4.2;乌梅丸多糖能降低结肠组织损伤程度。结论:多糖可能是乌梅丸汤剂中有效的活性成分,对小鼠UC有一定的防治作用,其理化性质分析方法简便快速,结果准确,重复性好,可作为乌梅丸多糖的质量控制方法。

The Physicochemical Properties of the Water Extract Polysaccharides ofWumei Pill and the Preventive Effect on DSS-induced Ulcerative Colitis

Objective:To determinate the polysaccharides content of water extract from Wu Mei pill and establish an HPLCanalysis method of evaluating the monosaccharide composition and molecular weight of polysaccharides from Wu Mei Pill. To observethe effect of the polysaccharides on mice with ulcerative colitis (UC) induced by dextran sulfate sodium (DSS).And to provide a basis forthe clinical use and the research on active ingredients of Wu Mei Pill.Methods:Water extraction and alcohol precipitation method wasused to extract polysaccharides. Sulfuric acid and phenol determination method was employed to detect the polysaccharides content ofWu Mei Pill. Gel chromatography and Long Zhida molecular weight work station were used to assess the molecular weight ofpolysaccharides. And PMP pre-column derivatization HPLC method was used to analyze the composition of monosaccharide. DSS (3 %)was used to make the UC mouse model. Fifty mice were randomly and equally divided into five groups, namely, normal group, modelgroup, and three doses of Wu Mei pill polysaccharides-treated groups (2.5 %, 5 %, and 10 %). The normal group was only given distilledwater, and the mice in other groups were given 3 %DSS for 7 days. Wu Mei pill polysaccharides were added in the water 7 days beforeDSS was given. All the mice were sacrificed in the 14th day.Results:Glucose in concentration range of 0 -0.08 mg·mL-1 presented agood linear relationship (r = 0.998); Dextran showed a good linear relationship in the molecular weight range of 2500 -2000000Da (r =0.9987). Polysaccharides content in water extract of Wu Mei Pill was 40.3 %. The sugar content of purified Wu Mei pill polysaccharideswas 91.6 %. The molecular weight ranged from 171343 to 525009 Da, with a coefficient of 1.18. The composition of monosaccharidewas mannose, glucuronic acid, glucose, galactose and xylose, with a relative ratio of 1.8:1.0:19.3:32.8:4.2. Wu Mei pill polysaccharidescould alleviate the inflammation in the colon.Conclusion:Polysaccharides, showing a moderate preventive and treatment effect on UC,may be one of the effective active ingredients of Wu Mei Pill decoction. The analytic method of chemical properties of Wu Mei pillpolysaccharides is simple, rapid, accurate, and reproducible and could be used as the quality control method.