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目的研究海带根中alpha-葡萄糖苷酶抑制剂的分离纯化与酶抑制作用动力学。方法采用85%乙醇溶液浸提、石油醚萃取、乙酸乙酯萃取、硅胶柱层析、Sephadex LH-20柱层析等方法,以alpha-葡萄糖苷酶抑制活性为导向,分离纯化得到了alpha-葡萄糖苷酶抑制剂活性组分;运用糖尿病小鼠模型探讨了海带根提取物体内降糖机理;采用双倒数作图法表征了活性组分抑制alpha-葡萄糖苷酶的动力学特征。结果海带根乙醇提取物能够有效降低糖尿病小鼠的餐后血糖;可显著抑制alpha-葡萄糖苷酶活性,IC50为13.6μg·mL-1;动力学研究表明酶抑制类型为非竞争性抑制。结论海带根提取物能够有效降低模型小鼠的血糖含量,其活性组分可显著抑制alpha-葡萄糖苷酶活性,对alpha-葡萄糖苷酶的抑制作用类型为非竞争性抑制。
Objective To study the isolation and purification of alpha-glucosidase inhibitors in kelp root and the kinetics of enzyme inhibition. METHODS: The alpha-glucosidase inhibitory activity was used as a guide for the extraction of alpha-glucosidase by using 85% ethanol solution, petroleum ether extraction, ethyl acetate extraction, silica gel column chromatography and Sephadex LH-20 column chromatography. Glucosidase inhibitor active components; diabetic mouse model was used to investigate the hypoglycemic mechanism of the kelp root extract; kinetic characteristics of the active component inhibition alpha-glucosidase were characterized by double reciprocal mapping. Results The kelp root ethanol extract could effectively reduce the postprandial blood glucose in diabetic mice, and could inhibit the alpha-glucosidase activity with IC50 of 13.6μg · mL-1. Kinetic studies showed that the type of enzyme inhibition was noncompetitive. Conclusion kelp root extract can effectively reduce the blood glucose in model mice, the active ingredient can significantly inhibit alpha-glucosidase activity, inhibition of alpha-glucosidase type non-competitive inhibition.