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基于 LC-QTOF-MS 的大籽猕猴桃果实代谢组学分析及α-葡萄糖苷酶抑制动力学评价。

LC-QTOF-MS-based metabolite profiling and evaluation of α-glucosidase inhibitory kinetics of Coccinia grandis fruit.

机构信息

School of Natural Product Studies, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India.

Institute of Bio-resources and Sustainable Development, A National Institute under Department of Biotechnology, Ministry of Science and Technology, Government of India, Imphal, India.

出版信息

Biomed Chromatogr. 2020 Dec;34(12):e4950. doi: 10.1002/bmc.4950. Epub 2020 Jul 29.

Abstract

Coccinia grandis is an important food crop of the Cucurbitaceae family, widely used for culinary purposes in India. It is reported to possess hypoglycemic, hypolipidemic and antioxidant activities. The current study was aimed to explore the inhibition kinetics as well as major constituents of the active fraction of C. grandis against α-glucosidase. The kinetic study was performed through spectrophotometric assay, with p-nitrophenyl-α-d-glucopyranoside as a substrate with varying concentrations. An in vitro antioxidant study was performed by DPPH assay. In addition, UPLC-QTOF-MS analysis was carried out for metabolite profiling of the bioactive fraction of C. grandis. The results showed that the difference between the α-glucosidase inhibitory activity of the ethyl acetate fraction of C. grandis (EFCG) (IC 2.43 ± 0.27 mg/ml), and standard inhibitor, acarbose (2.08 ± 0.19 mg/ml), was not statistically significant at a P-value of 0.05. The enzyme kinetics confirmed the inhibition mode in a mixed manner. The EFCG also showed the highest antioxidant activity (101.74 ± 1.95 μg/ml) among all of the fractions. A significant correlation between antioxidant and α-glucosidase inhibitory activity of EFCG was observed. The LC-QTOF-MS study of the EFCG putatively identified 35 metabolites, which may be responsible for its antioxidant and α-glucosidase inhibitory properties. Thus, C. grandis fruits can serve as a functional food to address diabetes-related disorders associated with α-glucosidase.

摘要

番荔枝是葫芦科的一种重要食用作物,在印度被广泛用于烹饪。据报道,它具有降血糖、降血脂和抗氧化活性。本研究旨在探索番荔枝活性部位对α-葡萄糖苷酶的抑制动力学及主要成分。动力学研究通过分光光度法进行,以 p-硝基苯-α-d-吡喃葡萄糖苷为底物,浓度不同。体外抗氧化研究采用 DPPH 法进行。此外,还进行了 UPLC-QTOF-MS 分析,以研究番荔枝生物活性部位的代谢产物谱。结果表明,番荔枝乙酸乙酯部位(EFCG)(IC 2.43±0.27 mg/ml)与标准抑制剂阿卡波糖(2.08±0.19 mg/ml)的α-葡萄糖苷酶抑制活性差异在 P 值为 0.05 时无统计学意义。酶动力学证实了抑制模式为混合模式。EFCG 还显示出所有馏分中最高的抗氧化活性(101.74±1.95 μg/ml)。EFCG 的抗氧化和α-葡萄糖苷酶抑制活性之间存在显著相关性。EFCG 的 LC-QTOF-MS 研究推测鉴定出 35 种代谢物,这些代谢物可能与其抗氧化和α-葡萄糖苷酶抑制特性有关。因此,番荔枝果实可作为一种功能性食品,用于治疗与α-葡萄糖苷酶相关的糖尿病相关疾病。

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