海藻铁钉菜抑制α-葡萄糖苷酶活性部位的筛选及其反应动力学研究
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摘要
目的:探讨海藻铁钉菜提取物对α-葡萄糖苷酶活性的影响及其反应动力学。方法:萃取法获得铁钉菜甲醇提取物的石油醚部位、二氯甲烷部位、乙酸乙酯部位、正丁醇部位、水部位等5个部位,采用PNPG筛选其中具有α-糖苷酶酶抑制的活性部位,通过酶促动力学方法与Lineweaver-Burk曲线,推断酶抑制类型。结果:包括甲醇提取物在内的6种铁钉菜提取物均具有一定的α-葡萄糖苷酶抑制活性,其中乙酸乙酯部位、石油醚部位、正丁醇部位、二氯甲烷部位的抑制活性均较强,IC50分别为3.4,4.4,5.1,7.8μg·mL~(-1),均远强于阳性对照阿卡波糖Acarbose(IC501 075.8μg·mL~(-1))。抑制动力学分析显示,乙酸乙酯部位、石油醚部位、正丁醇部位、二氯甲烷部位等对α-葡萄糖苷酶抑制作用均属于混合Ⅰ型抑制类型。结论:铁钉菜提取物具有作为新型α-葡萄糖苷酶抑制剂的药用开发价值,特别是乙酸乙酯部位。
OBJECTIVE To study the influence of seaweed Ishige okamurai onα-glucosidase activity and its reaction kinetics.METHODS Five solvent fractions,including petroleum ether fraction,dichloromethane fraction,ethylacetate fraction,n-butanol fraction and water fraction,were extracted from methanol extract of Ishige okamurai.Among them,α-glucosidase inhibiting extracts were screened by PNPG method,their inhibitory types were inferred by enzymatic dynamics and Lineweaver-Burk curve methods.RESULTS Six kinds of extracts,including methanol extract fromIshige okamurai,had certainα-glucosidase inhibiting activity,in which ethyl acetate fraction,petroleum ether fraction,dichloromethane fraction and n-butanol fraction had strong inhibitory activity with IC50 values of 3.4,4.4,5.1,7.8μg·mL~(-1),respectively,more potent than acarbose(IC501075.8μg·mL~(-1)),the positive control of this study.Inhibition dynamics analysis showed that the enzyme inhibitions of ethyl acetate fraction,petroleum ether fraction,dichloromethane fraction and n-butanol fraction were all close to mixed type Ⅰ inhibition.CONCLUSION The extracts of Ishige okamurai have medicinal development values as newα-glucosidase inhibitors,especially ethyl acetate fraction.
OBJECTIVE To study the influence of seaweed Ishige okamurai onα-glucosidase activity and its reaction kinetics.METHODS Five solvent fractions,including petroleum ether fraction,dichloromethane fraction,ethylacetate fraction,n-butanol fraction and water fraction,were extracted from methanol extract of Ishige okamurai.Among them,α-glucosidase inhibiting extracts were screened by PNPG method,their inhibitory types were inferred by enzymatic dynamics and Lineweaver-Burk curve methods.RESULTS Six kinds of extracts,including methanol extract fromIshige okamurai,had certainα-glucosidase inhibiting activity,in which ethyl acetate fraction,petroleum ether fraction,dichloromethane fraction and n-butanol fraction had strong inhibitory activity with IC50 values of 3.4,4.4,5.1,7.8μg·mL~(-1),respectively,more potent than acarbose(IC501075.8μg·mL~(-1)),the positive control of this study.Inhibition dynamics analysis showed that the enzyme inhibitions of ethyl acetate fraction,petroleum ether fraction,dichloromethane fraction and n-butanol fraction were all close to mixed type Ⅰ inhibition.CONCLUSION The extracts of Ishige okamurai have medicinal development values as newα-glucosidase inhibitors,especially ethyl acetate fraction.
引文
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[2]徐玉林,马东升,弓晓杰,等.5种稀有人参皂苷的制备及其对α-糖苷酶抑制活性的研究[J].河南农业大学学报,2012,46(3):317-321.
[3]Gao H,Huang YN,Gao B,et al.α-Glucosidase inhibitory effect by the flower buds of Tussilago farfara L.[J].Food Chem,2008,106(3):1195-1201.
[4]姚虹.α-葡萄糖苷酶抑制剂专利文献分析及其中药源新药开发进展[J].中国药业,2012,21(24):15-16.
[5]康文艺,张丽,宋艳丽.茜草抑制α-葡萄糖苷酶活性成分研究[J].中国中药杂志,2009,34(9):1104-1107.
[6]盛占武,王会,郑丽丽,等.香蕉花提取物中抑制α-葡萄糖苷酶活性组分研究[J].中国食品学报,2014,14(9):68-74.
[7]徐婷,褚江洪,丁萍英,等.葛根素对α-葡萄糖苷酶抑制作用机理的探讨[J].中国生化药物杂志,2015,35(1):43-45.
[8]李宪璀,范晓,韩丽君,等.海藻提取物中α-葡萄糖苷酶抑制剂的初步筛选[J].中国海洋药物,2002,21(2):8-11.
[9]汤海峰,易杨华,姚新生,等.褐藻铁钉菜化学成分的研究(Ⅱ)[J].中国海洋药物,2003,22(4):8-12.
[10]金鼎.海洋药物与效方[M].北京:中医古藉出版社,1998:90.
[11]汤海峰,易杨华,姚新生,等.褐藻铁钉菜中的甾醇成分[J].中国海洋药物,2002,21(1):1-4.
[12]龚丽芬,龚文贤.超声波协同热水提取铁钉菜多糖的研究[J].氨基酸和生物资源,2006,28(4):11-13.
[13]郑怡,余萍,刘燕如.3种海藻凝集素组分及糖抑制作用的初步研究[J].植物资源与环境学报,2002,11(4):19-21.
[14]汤海峰,易杨华,姚新生,等.铁钉菜化学成分的研究(3)[J].中国中药杂志,2002,27(4):269-273.
[15]Min KH,Kim HJ,Jeon YJ,et al.Ishige okamurae ameliorates hyperglycemia and insulin resistance in C57BL/KsJ-db/db mice[J].Diabetes Res Clin Pract,2011,93(1):70-76.
[16]张丽.滇丁香、茜草抑制α-葡萄糖苷酶活性成分研究[D].河南大学,2010.
[17]Li T,Zhang XD,Song YW,et al.A microplate-based screening method for alpha-glucosidase inhibitors[J].Chin J Clin Pharm Ther,2005,10(10):1129-1134.
[18]陈清西.酶学及其研究技术[M].厦门:厦门大学出版社,2010:120.
[19]张耀东,蔡亚楠,张琦,等.淫羊藿苷对乙酰胆碱酯酶的抑制作用[J].药学学报,2012,47(9):1141-1146.
[20]王翼,张旭.α-葡萄糖苷酶抑制剂的研究进展[J].海峡药学,2009,21(9):4-6.
[21]马燕燕,鲁晓翔.天然产物α-葡萄糖苷酶抑制剂筛选研究进展[J].粮食与油脂,2010,6:7-9.