摘要
目的从大黄中提取多糖,并测定提取物粗粉中大黄多糖的含量,研究大黄多糖对酶-抑制剂模型和Caco-2细胞模型上α-葡萄糖苷酶的抑制作用,初步探索大黄多糖在治疗糖尿病方面的潜在价值。方法用水提醇沉法提取大黄多糖,用苯酚-硫酸比色法测定多糖含量。采用氧化酶比色法测定大黄多糖对α-葡萄糖苷酶(EC3.2.1.20)以及Caco-2细胞模型表达α-葡萄糖苷酶的抑制活性。结果大黄多糖粗粉中大黄多糖平均含量为(23.08±0.89)%,RSD=3.87%(n=3),方法平均回收率为97.59%,RSD=3.15%(n=6)。以麦芽糖为底物时,在酶-抑制剂模型上,浓度为125,250,500,1000μg·mL-1的大黄多糖的抑制率分别为33.09%,24.43%,33.14%,31.73%,250μg·mL-1的阳性药阿卡波糖的抑制率为80.45%。在Caco-2细胞模型上,浓度为125,250,500,1000μg·mL-1的大黄多糖抑制率分别为23.43%,26.23%,28.24%,29.17%,250μg·mL-1的阿卡波糖对Caco-2细胞的α-葡萄糖苷酶抑制率达到84.08%。结论运用水提醇沉法提取大黄多糖,用酚-硫酸比色法测定多糖含量方法简便可行,大黄多糖具有一定的抑制α-葡萄糖苷酶活性的作用,但作用不显著。
Objective To extract polysaccharide from rhubarb,determine the content and study the α-glucosidase inhibition effect on the enzyme-inhibitor model and Caco-2 cells model for preliminary exploring the potential value of rhubarb polysaccharides on treating diabetes. Methods Rhubarb polysaccharides were extracted by water isolation and alcohol precipitation,and the content was determined with phenol-sulfuric acid colorimetry. The inhibitory activity of rhubarb polysaccharides on α-glucosidase （ EC 3. 2. 1. 20） was measured,as well as α-glucosidase expressed on Caco-2 cell model by using oxidase colorimetric method. Results The average concentration of rhubarb polysaccharides in rhubarb extracts was （ 23. 08 ± 0. 89） %,RSD = 3. 87% （ n = 3） ,and average recovery was 97. 59%,RSD = 3. 15% （ n = 6） . With maltose as the substrate in the enzyme-inhibitor model,the inhibition rates of 125,250,500,1 000 μg·mL-1 rhubarb polysaccharides were 33. 09% ,24. 43% ,33. 14% ,31. 73% ,respectively. The inhibition rate of 250 μg·mL-1 positive drug （ acarbose ） was 80. 45% . On the Caco-2 cell model,the inhibition rates of 125,250,500,1 000 μg·mL-1rhubarb polysaccharides were 23. 43% , 26. 23% ,28. 24% ,29. 17% ,and 250 μg·mL-1 positive drug （ acarbose） reached 84. 08% . Conclusion Water extraction and alcohol precipitation for Rhubarb polysaccharides isolating and phenol-sulfuric acid colorimetric assay for determining content of polysaccharide are simple and feasible. Rhubarb polysaccharides has a certain degree of α-glucosidase inhibitory activity.
出处
《医药导报》
CAS
2010年第8期985-989,共5页
Herald of Medicine
基金
陕西省自然科学基金资助项目(基金编号:2005K18-Z9)
关键词
大黄多糖
含量测定
α-葡萄糖苷酶抑制作用
Rhubarb Polysaccharide
Content determination
α-Glucosidase inhibition
