| 376 | 7 | 24 |
| 下载次数 | 被引频次 | 阅读次数 |
目的研究酵母和大鼠小肠黏膜来源的α-葡萄糖苷酶活性测定的体外体系,评价在该体系条件下表没食子儿茶素没食子酸酯(EGCG)的α-葡萄糖苷酶抑制活性(AGI)的活性,为建立AGI活性评价体系提供依据。方法分别应用酵母和大鼠小肠黏膜提取液作为α-葡萄糖苷酶,以麦芽糖为底物;用0.00、0.25、0.50、1.0和1.50mg/ml的酶与0.00、0.01、0.02、0.04、0.06、0.08、0.1和0.12mol/l的底物反应,采用葡萄糖氧化酶法测定葡萄糖生成量来确定反应条件;在该反应条件下比较两种酶源的活性差异,并以此分析EGCG的AGI作用和计算半数抑制浓度(IC50)。结果最佳反应条件为0.50mg/ml的酶与0.06mol/l的底物反应,研究发现在该体系下EGCG对酵母来源的AGI活性强于小肠黏膜来源的酶,其IC50值分别为:0.150和0.837mg/ml。结论建立体外AGI活性评价体系有利于准确筛选AGI活性物质,酵母来源的酶可用于AGI活性的初步筛选;离体组织测定法可用于进一步评价AGI活性物质;EGCG体外具有较强的AGI活性。
Abstract：Objective To investigate the effect of epigallocatechin-3-gallate ( EGCG) on the activity of α- glucosidase derived from bakers’yeast and small intestinal mucosa of Wistar rats,in order to establish an evaluation system for the activity of α-glucosidase inhibitor. Methods α-glucosidase was prepared from the bakers’yeast and the small intestinal mucosa of Wistar rats. The inhibitory activity of EGCG on α-glucosidase was based on the concentration of glucose in the reaction system. Results EGCG was shown to be an α-glucosidase inhibitor. IC50 was 0. 150mg /ml and 0. 837mg /ml for α-glucosidase derived from bakers ’ yeast and small intestinal mucosa respectively. Conclusion The establishment of an in vitro evaluation system for AGI activity is conductive for screening α-glucosidase inhibitory substance. The AGI activity of EGCG is strong in vitro.
1KIM J S,KWON C S,SON K H.Inhibition of alpha-glucosidase and amylase by luteolin,a flavonoid[J].Biosci Biotechnol Biochem,2000,64(11):24581.
2FLORIS A,PETER L,REINIER P,et al.α-glucosidase inhibitors for patients with type2diabetes[J].Diabetes Care,2005,28(1):154-163.
3LEVETTAN C.Oral antidiabetic agents in type2diabetes[J].Curr Med Res Opin,2007,23(4):945-952.
4YEH G Y,EISENBERG D M,KAPTCHUK T J,et al.Systematic review of herbs and dietary supplements for glycemic control in diabetes[J].Diabet Care,2003,26(4):1277-1294.
5王竹,杨月欣.植物化学物α-葡萄糖苷酶抑制剂样降血糖作用及其评价方法[J].国外医学卫生学分册,2008,35(6):355-358.
6PIERRE C,TREMBLAY R R,DUBE J Y.P-nitrophenol-α-glucopyranoside as substrate for measurement of maltase activity in human semen[J].J Clinic Chem,1978,24:208-211.
7全吉淑,尹学哲,张帅.大豆胚芽提取物体外对小肠黏膜蔗糖酶、麦芽糖酶及葡萄糖转运活性的影响[J].食品科学,2004,25(1):161-163.
8王镜岩,朱圣庚,徐长法.生物化学[M].北京:高等教育出版社,2002.
9RORAERO A P,PETER G J,SHA Hinian,et al.The yeast CWH41gene encodes glucosidase I[J].Dycobiology,1997,7(7):997-1004.
10徐雯,曹艳华,乔德水,等.α-葡萄糖苷酶三维结构的同源模建及其对接研究[J].药物生物技术,2009,16(3):255-259.
11周敏,王正祥.黑曲CICIM F0410中α-葡萄糖苷酶的酶学性质研究[J].生物技术通报,2009,1:130-135.
12HUNZIKER W,PIESS S M,SEMENZA G,et al.The sucrase-isomaltase complex:primary structure,membrane-orientation,and evolution of a stalked,intrinsic brush border protein[J].Cell,1986,46(2):227-234.
13ODAKA H,MIKI N,IKEDA H,et al.Effect of a disaccharidase inhibitor,AO2128,on post-prandial hyperglycemia in rats[J].J Jpn Soc Nutr Food Sci,1992,45(1):27-31.
14全吉淑,尹学哲,张帅.绿茶提取物降血糖机制研究[J].食品科学,2008,25(1):21-25.
15李英,沈忠明.α-葡萄糖苷酶抑制剂的分离纯化及其性质研究[J].天然产物研究与开发,2000,12(4):24-29.
16中华人民共和国农业部.NY/T1103.2-2006转基因植物及其产品食用安全检测抗营养素第2部分:胰蛋白酶抑制剂的测定[S].北京:中华人民共和国农业部,2006.
17陈海敏,严小军,林伟.α-葡萄糖苷酶抑制剂的构效关系[J].中国生物化学与分子生物学报,2003,19(6):780-784.
基本信息:
DOI：10.19813/j.cnki.weishengyanjiu.2010.02.012
中图分类号:Q55
引用信息:
[1]祝宇铭,丁红,王竹,等.表没食子儿茶素没食子酸酯体外α-葡萄糖苷酶活性的影响[J].卫生研究,2010,39(02):168-171+176.DOI:10.19813/j.cnki.weishengyanjiu.2010.02.012.
基金信息:
国家“十一五”科技支撑计划项目(No.2006BAD27B01)
2010-03-30
2010-03-30