目的 探索山楂Crataegi Fructus干燥果实的不同炮制品提取物抑制α-葡萄糖苷酶活性差异以及成分变化规律。方法 生山楂：将山楂切薄片置于烘箱中烘干至恒定质量（60 ℃、16 h）；炒山楂：将生山楂置于烤箱中加热至表皮呈黄色、果肉黄褐色（80 ℃、20 min；100 ℃、15 min）；焦山楂：将生山楂烤至表面焦褐色、内部黄褐色，并具有焦香气味（120 ℃、20 min；150 ℃、15 min）。将生、炒、焦山楂用粉碎机粉碎至粉末，过60目筛，−80 ℃储存备用。通过体外α-葡萄糖苷酶活性抑制模型以及动物模型评价山楂不同炮制品提取物对高血糖的抑制作用。基于UPLC-Q-TOF-MS/MS结合线上数据库（GNPS、MSDIAL、PubChem等）对山楂不同炮制品化学成分进行鉴定，明确炮制工艺对山楂中化学成分的影响。结果 体外酶活研究结果显示，山楂炒制后其提取物的α-葡萄糖苷酶的抑制活性显著增强（P＜0.05），生、炒、焦山楂提取物以及阿卡波糖半数抑制浓度（median inhibition concentration，IC₅₀）分别为（28.31±2.57）、（12.67±1.01）、（928.33±131.02）、（265.50±31.50）μg/mL；动物实验表明，炒山楂提取物能够显著改善餐后高血糖、葡萄糖耐受性、提高胰岛素敏感性。利用LC-MS/MS结合GNPS对山楂不同炮制品的成分进行分析，参考PubChem、Massbank等公共数据库及文献，共鉴定59种化合物，主要含有黄酮糖苷类、原花青素类、三萜类等3大类化合物。半定量结果表明，山楂在炒制过程中化学成分变化显著，其中大部分黄酮类、三萜类成分相对含量升高，原花青素类成分经炒制后含量相对降低。结论 山楂炒制后其α-葡萄糖苷酶抑制活性显著增强，动物实验表明炒山楂提取物具有良好的抑制高血糖的作用。山楂炒制过程化学成分发生显著变化，这可能是炒山楂提取物α-葡萄糖苷酶抑制活性增强的重要原因，为阐明炒山楂的降血糖机制以及炒山楂降糖产品开发提供了理论依据。

Objective To explore the differences in α-glucosidase inhibitory activity and the changing rules of components in extracts from different processed products of the dried fruits of Shanzha (Crataegi Fructus).. Methods Raw Crataegi Fructus: Crataegi Fructus slices are placed in the oven to dry to constant weight (60 ℃, 16 h). Stir-fried Crataegi Fructus: Place raw Crataegi Fructus in the oven and heat until the skin is yellow and the flesh is yellowish brown (80 ℃, 20 min and 100 ℃, 15 min). Burnt Crataegi Fructus: heat raw Crataegi Fructus until the surface is burnt brown, the interior is yellowish brown, and has a burnt odor (120 ℃, 20 min and 150 ℃, 15 min). The raw, fried and scorched Crataegi Fructus were crushed to powder with a grinder, passed through a 60-mesh sieve, and stored at −80 ℃ for later use. The inhibitory effects of extracts from different Crataegi Fructus products on hyperglycemia were evaluated by α-glucosidase activity inhibition model in vitro and animal model in vivo. Based on UPLC-Q-TOF-MS/MS combined with online databases (GNPS, MSDIAL, PubChem, etc.), the chemical components of different Crataegi Fructus products were identified, and the effects of processing technology on the chemical components of Crataegi Fructus were clarified. Results The results of in vitro enzyme activity study showed that the α-glucosidase inhibitory activity of Crataegi Fructus extract was significantly enhanced after stir-fry (P < 0.05) (IC₅₀＝28.31±2.57 μg/mL in raw Crataegi Fructus, IC₅₀＝12.67±1.01 μg/mL in fried Crataegi Fructus, IC₅₀＝928.33±131.02 μg/mL in burnt Crataegi Fructus, IC₅₀＝265.50±31.50 μg/mL in acarose); Animal experiments showed that stir fried hawthorn extract could improve postprandial hyperglycemia, glucose tolerance, and insulin sensitivity. LC-MS combined with GNPS were used to analyze the components of different Crataegi Fructus products, and 59 compounds were identified with reference to PubChem, Massbank and other public databases and literature, mainly containing three categories of compounds, namely flavonoid glycosides, proanthocyanidins and triterpenoids. The semi-quantitative results showed that the chemical components of Crataegi Fructus changed significantly during the roasting process, and the relative contents of most flavonoids and triterpenoids increased, while the contents of proanthocyanidins decreased after roasting. Conclusion The inhibitory activity of α-glucosidase activity of Crataegi Fructus was significantly enhanced after stir-frying. Animal experiments showed that stir-fried Crataegi Fructus extract had a good inhibitory effect on hyperglycemia. The stir-frying process of Crataegi Fructus has a great influence on the chemical composition, which can be the basis for stir-frying Crataegi Fructus to inhibit and enhance the activity of α-glucosidase. This study provides a theoretical basis for the mechanism of lowering blood glucose in stir-fried Crataegi Fructus and the development of stir-fried Crataegi Fructus lowering sugar products.

R284.1；R285

浙江中医药大学自然科学青年探索项目（2024JKZKTS05）