食用海藻中α-葡萄糖苷酶抑制剂的筛选及抑制动力学
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摘要
为探讨广西食用海藻不同极性部位提取物对α-葡萄糖苷酶活性的影响及抑制作用动力学。采用萃取法制得食用海藻(海带、裙带菜、紫菜)的甲醇部位、石油醚部位、乙酸乙酯部位、正丁醇部位及水部位,采用体外筛选(PNPG)法筛选抑制α-葡萄糖苷酶活性的部位。通过酶促反应动力学研究高抑制活性部位的反应动力学,绘制Lineweaver-Burk曲线,判断抑制剂与酶作用的抑制类型。结果表明,广西北部湾食用海藻(海带、裙带菜、紫菜)的不同极性部位均有一定的α-葡萄糖苷酶抑制活性,其中紫菜的乙酸乙酯部位抑制活性相对较高,其半数抑制浓度(IC_(50))为7.89 mg/mL,其抑制活性高于阳性对照阿卡波糖(IC_(50)=11.45 mg/mL)。食用海藻(海带、裙带菜、紫菜)的乙酸乙酯部位的抑制动力学研究结果表明对α-葡萄糖苷酶抑制作用属于混合型抑制类型。广西食用海藻的不同极性部位均具有一定的α-葡萄糖苷酶抑制活性,有望开发为新型α-葡萄糖苷酶抑制剂。
The influence of edible algal of Guangxi on α-glucosidase activity and its inhibition kinetic behaviors were studied.Five solvent fractions were prepared by extraction from edible algal, including methanol fraction, petroleum ether fraction,ethyl acetate fraction, n-butyl alcohol fraction and water fraction. The α-glucosidase inhibiting extracts were screened by vitro screening(PNPG) method. The kinetics of high inhibitory activity sites was studied by enzyme-catalyzed reaction, and the lineweaver-burk curve was drawn to deduce their inhibitory types. Results showed that five kinds of extracts from edible algal had certain α-glucosidase inhibiting activity, in which ethyl acetate fraction of nori had strong inhibitory activity with IC_(50) value of 7.89 mg/mL. Its inhibitory activity was higher than acarbose(IC_(50)=11.45 mg/mL) in the positive control of this study. Inhibition dynamics analysis showed that the enzyme inhibitions of ethyl acetate fraction of edible algal were all close to mixed type Ⅰ inhibiton. The different extracts of edible algal in guangxi had certain inhibitory activity and the medical development values as a new α-glucosidase inhibitor, especially ethyl acetate fraction of nori.
The influence of edible algal of Guangxi on α-glucosidase activity and its inhibition kinetic behaviors were studied.Five solvent fractions were prepared by extraction from edible algal, including methanol fraction, petroleum ether fraction,ethyl acetate fraction, n-butyl alcohol fraction and water fraction. The α-glucosidase inhibiting extracts were screened by vitro screening(PNPG) method. The kinetics of high inhibitory activity sites was studied by enzyme-catalyzed reaction, and the lineweaver-burk curve was drawn to deduce their inhibitory types. Results showed that five kinds of extracts from edible algal had certain α-glucosidase inhibiting activity, in which ethyl acetate fraction of nori had strong inhibitory activity with IC_(50) value of 7.89 mg/mL. Its inhibitory activity was higher than acarbose(IC_(50)=11.45 mg/mL) in the positive control of this study. Inhibition dynamics analysis showed that the enzyme inhibitions of ethyl acetate fraction of edible algal were all close to mixed type Ⅰ inhibiton. The different extracts of edible algal in guangxi had certain inhibitory activity and the medical development values as a new α-glucosidase inhibitor, especially ethyl acetate fraction of nori.
引文
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[19] LI Y Q, ZHOU F C, GAO F. Comparative evaluation of quercetin, isoquercetin and rutin as inhibitors ofα-glucosidase[J].Journal of Agricultural and Food Chemistry, 2009, 57(24):11463-11467.
[2]余颖,樊金玲,程源斌,等.甘草酸提取废液α-葡萄糖苷酶抑制剂的筛选与鉴定[J].食品科学, 2018, 39(13):22-28.
[3]张红霞.口服降糖药物不良反应研究进展[J].天津药学,2017, 29(2):75-78.
[4]延海莹,刘盟梦,乔乐克,等.扇贝裙边活性肽的制备及其降血糖活性研究[J].食品工业, 2018, 39(3):117-121.
[5]陈哲,张进,钟振声.金线莲α-葡萄糖苷酶抑制剂提取工艺优化研究[J].中国酿造, 2017, 36(7):135-13.
[6]谭哲谞,许利嘉,陈四保.植物来源α-葡萄糖苷酶抑制剂的研究新进展[J].中南药学, 2018(7):1-5.
[7]熊能,韦圣,胡忠策.α-葡萄糖苷酶抑制剂的研究进展和趋势[J].发酵科技通讯, 2018, 47(1):58-64.
[8]王凯.α-葡萄糖苷酶抑制剂的来源、筛选模型及临床研究进展[J].中国药房, 2014, 25(41):3919-3922.
[9]季芳,肖国春,董莉,等.药用植物来源的α-葡萄糖苷酶抑制剂研究进展[J].中国中药杂志, 2010, 35(12):1633-1640.
[10]周锋,刘冬冬,周楠迪,等.条斑紫菜蛋白酶解产物中α-葡萄糖苷酶抑制剂纯化及特性[J].天然产物研究与开发,2014, 26(9):1463-1468.
[11]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(2):8-11.
[12]黄晓冬,蔡琼瑶,王远花,等.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[J].中国医院药学杂志, 2016, 36(1):16-20.
[13]周锋,刘冬冬,周楠迪,等.条斑紫菜蛋白酶解产物中α-葡萄糖苷酶抑制剂纯化及特性[J].天然产物研究与开发,2014, 26(9):1463-1468.
[14]LIUQ,HUHJ,LIOF,etal.Diterpenoidsand phenylethanoid glycosides from the roots of Clerodendrum bungei and their inhibitory effects Against angiotensin converting enzyme andα-glucosidase[J]. Phytochemistry,2014, 103:196-202.
[15]李晨悦,李冬利,孙玉坤,等.簕菜提取物及其化学成分对α-葡萄糖苷酶的抑制作用[J].食品工业, 2016, 37(4):164-167.
[16]朱娟娟,尹忠平,陈继光,等.微量α-葡萄糖苷酶抑制剂筛选模型及抑制类型的判断方法[J].现代食品科技, 2016,32(12):164-170, 157.
[17]杨荣武.生物化学原理[M].北京:高等教育出版社, 2012:132-149.
[18]杨文康,李超,喻柯柯,等.罗汉果皂甙粗提物的α-葡萄糖苷酶体外抑制活性研究[J].食品工业科技, 2016, 37(24):111-115.
[19] LI Y Q, ZHOU F C, GAO F. Comparative evaluation of quercetin, isoquercetin and rutin as inhibitors ofα-glucosidase[J].Journal of Agricultural and Food Chemistry, 2009, 57(24):11463-11467.