山莓叶中有效成分的分离鉴定及其生物活性研究

Isolation and Identification of Active Ingredients in Rubus corchorifolia L.f. Leaves and Their Bioactivities

摘要:

本文以山莓叶乙醇提取物为研究对象，通过溶剂萃取、硅胶柱层析、结晶等方法进行分离纯化。采用核磁共振（NMR），质谱（MS）、红外光谱法（IR）及与文献对比的方法对其分离的单体进行结构鉴定。利用刃天青96孔板微量稀释法、MTT法研究单体化合物的抑菌及抗肿瘤活性。研究结果表明：从山莓叶中分离纯化物经结构鉴定分别为：三萜化合物类2α、3β、23α-三羟基-12-烯-28-乌苏酸（1）和黄酮类化合物山奈酚-3-O-β-D-（6’’-对羟基桂皮酰基）-葡萄糖苷（2）；抑菌实验表明化合物1对供试的大肠杆菌、痢疾志贺氏菌、金黄色葡萄球菌、沙门氏菌最低抑菌浓度分别为：15.6 μg/mL、15.6 μg/mL、31.25 μg/mL、15.6 μg/mL，化合物2的最低抑菌浓度分别为：31.25 μg/mL、62.5 μg/mL、62.5 μg/mL、62.5 μg/mL。化合物1对HepG2、MCF-7、OVCAR3的半数抑制率（IC50）分别为：34.2 μM、60.43 μM、62.3 μM。化合物1具有很好的抑菌和较好的抗肿瘤活性。而化合物2对HepG2、MCF-7、OVCAR3细胞的抑制作用不明显。

Abstract:

The ethanol extract of Rubus corchorifolia leaves was used in this study, and its active compounds were isolated and purified using solvent extraction, silica gel column chromatography, and crystallization methods. The structures of the isolated monomeric compounds were identified using nuclear magnetic resonance (NMR), mass spectrometry (MS), infrared (IR) spectroscopy, and comparisons with literature data. The antibacterial and antitumor activities of the monomeric compounds were studied by resazurin microdilution (96-well plate) assay and MTT assay. The two compounds isolated and purified from Rubus corchorifolia leaves were identified as 2α,3β,23α-trihydroxy-12-en-28-ursolic acid (1) and flavonoid compound kaempferol-3-O-β-D-6′′-p-hydroxycoumaroyl-glucoside (2). The minimum inhibitory concentrations (MICs) of compound 1/compound 2 on Escherichia coli, Shigella dysenteriae, Staphylococcus aureus, and Salmonella were 15.6 μg/mL/31.25 μg/mL, 15.6 μg/mL/62.5 μg/mL, 31.25 μg/mL/62.5 μg/mL, and 15.6 μg/mL/62.5 μg/mL, respectively. The half inhibitory concentration (IC50) of compound 1 on HepG2, MCF-7, and OVCAR3 cells was 34.2 μM, 60.43 μM, and 62.3 μM, respectively. Compound 1 showed good antibacterial and antitumor activities, while the inhibitory effects of compound 2 on HepG2, MCF-7, and OVCAR3 cell lines were not significant.