胡芦巴生品、酒制品抑制α-葡萄糖苷酶和α-淀粉酶活性的有效部位筛选及其酶动力学分析

Screening of Effective Fraction from Raw and Rice-wine Products of Trigonellae Semen with Inhibiting Activity Against α-Glucosidase andα-Amylase and Analysis of Its Inhibition Kinetics

目的:比较胡芦巴生品和酒制品对α-葡萄糖苷酶,α-淀粉酶抑制活性作用的差别,筛选抑制活性的有效部位,并研究其酶反应动力学。方法:利用萃取法获得胡芦巴生品和酒制品80%甲醇提取物的石油醚层、乙酸乙酯层、正丁醇层、水层4个不同极性部位,采用体外α-葡萄糖苷酶抑制模型和α-淀粉酶抑制模型,测定生胡芦巴和酒胡芦巴4个不同极性部位对α-葡萄糖苷酶,α-淀粉酶活性的抑制作用。通过酶促动力学与Lineweaver-Burk曲线推断各有效部位对α-葡萄糖苷酶及α-淀粉酶抑制类型。结果:生胡芦巴和酒胡芦巴只有水层对α-葡萄糖苷酶有抑制作用,酒制品对α-葡萄糖苷酶的抑制作用优于生品,生胡芦巴和酒胡芦巴水层萃取物对α-葡萄糖苷酶作用为非竞争可逆。生胡芦巴和酒胡芦巴的乙酸乙酯层和正丁醇层对α-淀粉酶有抑制作用,酒制品的抑制作用优于生品,且均为非竞争可逆。结论:胡芦巴经酒制后可在一定程度上增加对α-葡萄糖苷酶及α-淀粉酶活性的抑制作用,酒胡芦巴水层萃取物有开发成α-葡萄糖苷酶抑制剂的价值,酒胡芦巴的乙酸乙酯和正丁醇提取物具有开发成α-淀粉酶抑制剂的价值。

Objective： To compare the inhibition effect of raw and rice-wine products of Trigonellae Semen on α-glucosidase and α-amylase for screening of effective fraction with inhibiting activity. Then try to analyze its reaction kinetics. Method： Four different polar fractions,including petroleum ether fraction,ethyl acetate fraction,n-butanol fraction and water fraction,were extracted from 80% methanol extract of raw and ricewine products of Trigonellae Semen. α-Glucosidase inhibitory model and α-amylase inhibition model in vitro were adopted to measure inhibition effect of different polar fractions on α-glucosidase and α-amylase. Then their inhibitory types were inferred by enzymatic dynamics and Lineweaver-Burk curve methods. Result： The water fraction from raw and rice-wine products of Trigonellae Semen had inhibition effect on α-glucosidase with noncompetitive and reversible type,the rice-wine product of Trigonellae Semen was better than raw product. The ethyl acetate and n-butanol fraction from raw and rice-wine products of Trigonellae Semen had inhibition effect onα-amylase with non-competitive and reversible type,and the rice-wine product of Trigonellae Semen was better than raw product. Conclusion： The inhibition effect of Trigonellae Semen on α-glucosidase and α-amylase can be increased after being processed by rice-wine. The water fraction from rice-wine product has medicinal development values as a new α-glucosidase inhibitor, the ethyl acetate and n-butanol fraction from rice-wine product has medicinal development values as new α-amylase inhibitors.