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Archive > Volume 16 Issue 1 > 2024,16(1):151-161. DOI:https://doi.org/10.1016/j.chmed.2022.11.009 Prev Next

文冠果花的降糖活性研究及基于自由基-UPLC-ESI-QTOF/MS離線聯(lián)用技術(shù)快速篩選文冠果花抗氧化活性成分

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Hypoglycemic activities of flowers of Xanthoceras sorbifolia and identification of anti-oxidant components by off-line UPLC-QTOF-MS/MS-free radical scavenging detection

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    摘要:
    目的：本研究的主要目的是分析文冠果花中活性部位的化學(xué)成分，并評價其抗氧化和抗高血糖的能力，確定主要活性成分。 方法：采用AlCl3比色法和普魯士藍(lán)法測定文冠果花提取物各萃取層中總黃酮和總酚酸含量，并通過體外抗氧化實驗篩選文冠果花中的生物活性部位。采用Waters ACQUITY UPLC BEH C18柱（2.0 mm×150 mm，5 μm），分析文冠果花中生物活性部位的化學(xué)成分，通過自由基-UPLC-ESI-QTOF/MS離線聯(lián)用技術(shù)快速篩選其抗氧化活性成分，評估其在高脂鏈脲佐菌素誘導(dǎo)的2型糖尿病大鼠模型中的作用，并且采用分子對接技術(shù)驗證提取物中作用于NADPH氧化酶（PDB ID: 2CDU）的主要活性物質(zhì)。 結(jié)果：結(jié)合體外活性篩選的結(jié)果，確定文冠果花的抗氧化部位為乙酸乙酯層。采用液相色譜-質(zhì)譜法在文冠果花的乙酸乙酯層中共鑒定出24種化學(xué)成分，并通過自由基-UPLC-ESI-QTOF/MS離線聯(lián)用技術(shù)初步篩選出13種主要抗氧化活性成分。根據(jù)體內(nèi)實驗結(jié)果發(fā)現(xiàn)文冠果花能顯著降低糖尿病小鼠血糖水平，減輕肝細(xì)胞損傷。根據(jù)分子對接的分析結(jié)果發(fā)現(xiàn)槲皮素3-O-蕓香糖苷、山奈酚-3-O-鼠李糖苷、異鼠李素-3-O-葡萄糖苷、異槲皮素對NADPH氧化酶表現(xiàn)出較好的抑制作用。 結(jié)論；文冠果花的乙酸乙酯層含有豐富的總黃酮和總酚酸成分，具有明顯的抗氧化活性，預(yù)防高血糖的發(fā)生。本文的研究表明文冠果花是一種有發(fā)展前景的新型茶源，可開發(fā)安全性高的天然抗糖尿病的替代產(chǎn)品。

    Abstract:
    Objective: To identify phytochemical constituents present in the extract of flowers of Xanthoceras sorbifolia and evaluate their anti-oxidant and anti-hyperglycemic capacities. Methods: The AlCl3 colorimetric method and Prussian Blue assay were used to determine the contents of total flavonoids and total phenolic acids in extraction layers, and the bioactive layers was screened through anti-oxidative activity in vitro. The Waters ACQUITY UPLC system and a Waters ACQUITY UPLC BEH C18 column (2.0 mm × 150 mm, 5 μm) were used to identify the ingredients. And antioxidative ingredients were screened by off-line UPLC-QTOF-MS/MS-free radical scavenging. The ameliorative role of it was further evaluated in a high-fat, streptozotocin-induced type 2 diabetic rat model and the study was carried out on NADPH oxidase (PDB ID: 2CDU) by molecular docking. Results: Combined with the results of activity screening in vitro, the anti-oxidative part was identified as the ethyl acetate layer. A total of 24 chemical constituents were identified by liquid chromatographymass spectrometry in the ethyl acetate layer and 13 main anti-oxidative active constituents were preliminarily screened out through off-line UPLC-QTOF-MS/MS-free radical scavenging. In vivo experiments showed that flowers of X. sorbifolia could significantly reduce the blood glucose level of diabetic mice and alleviate liver cell damage. Based on the results of docking analysis related to the identified phytocompounds and oxidase which involved in type 2 diabetes, quercetin 3-O-rutinoside, kaempferol-3-Orhamnoside, isorhamnetin-3-O-glucoside, and isoquercitrin showed a better inhibitory profile. Conclusion: The ethyl acetate layer was rich in flavonoids and phenolic acids and had significant antioxidant activity, which could prevent hyperglycemia. This observed activity profile suggested X. sorbifolia flowers as a promising new source of tea to develop alternative natural anti-diabetic products with a high safety margin.

    關(guān)鍵詞:
    抗高血糖;抗氧化劑;文冠果花;NADPH氧化酶;自由基-UPLC-QTOF-MS/MS離線聯(lián)用技術(shù)

    Keywords:


    Project Supported:
    The financial support of this study was from the Fourth National Investigation of Chinese materia medica resources in Liaoning Province (LN2018017, LN2019019); Career Development Support Plan for Young and Middle-aged Teachers in Shenyang Pharmaceutical University (No. ZQN2021014); and Key Laboratory of Marine Biogenetic Resources, Ministry of Natural Resources (No. HY202105).

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