CAJ | 학술논문

目的:建立敏感的微孔板方法检测alpha-葡萄糖苷酶抑制活性,以进行抑制剂体外高通量筛选.方法:调整酶和底物适当配比,通过标准曲线、波长扫描、动力学分析以及最佳反应条件的研究,用阿卡波糖验证所建方法.结果:确定反应步骤,96孔板,160 μl体系含 2.5 mmol·L-1 PNPG和 0.2 U·ml-1 alpha-葡萄糖苷酶,37 ℃,pH 7.0 反应15 min,400 nm检测,此法正确、可靠.结论:本文所述用小体积样品筛选得到大量抑制剂的方法,利于从天然产物中开发改善餐后血糖的糖尿病及其并发症治疗药物.
목적:건립민감적미공판방법검측alpha-포도당감매억제활성,이진행억제제체외고통량사선.방법:조정매화저물괄당배비,통과표준곡선、파장소묘、동역학분석이급최가반응조건적연구,용아잡파당험증소건방법.결과:학정반응보취,96공판,160 μl체계함 2.5 mmol·L-1 PNPG화 0.2 U·ml-1 alpha-포도당감매,37 ℃,pH 7.0 반응15 min,400 nm검측,차법정학、가고.결론:본문소술용소체적양품사선득도대량억제제적방법,리우종천연산물중개발개선찬후혈당적당뇨병급기병발증치료약물.
AIM: To establish a new sensitive microplate-based method to determine alpha-glucosidase inhibiting activity and provide a reliable high-throughput way for screening alpha-glucosidase inhibitors in vitro.METHODS: The fitting combination of enzyme and substrate in a certain reaction was tested.Acarbose,the most popular alpha-glucosidase inhibitor in clinical use was used to validate the established method.Calibration curve,wavelength fidelity and kinetic analysis,together with the effect of altered incubation time,temperature and pH were then studied.RESULTS:The details of assay procedure and evaluation of factors affecting the measurement are described.As little as 160 μl assay system was performed in a 96-well plate.The optimal action was finally achieved by incubated at 37 ℃,pH 7.0 for 15 min and measured at 400 nm.Results from the validation exercises by Acarbose strongly demonstrated the accuracy and reliability of the proposed approach.CONCLUSION: This method reported in the current paper makes it possible to rapidly examine large numbers of samples for the presence of alpha-glucosidase inhibitors in very small sample volumes.Such action may help to pace the development of potential oral medications from natural products protecting patients against postprandial hyperglycemic toxicity and therefore treating diabetes mellitus and the related complications.