油橄榄果提取物体外降脂活性成分筛选及作用机制探究

Screening of in vitro lipid-lowering active ingredients from Olea europaea L.fruit extract and preliminary study on the mechanism

目的 探究油橄榄果提取物体外降脂活性物质基础,并初步探讨活性成分在体内的降脂作用机制。方法 建立17个油橄榄果提取物样品高效液相色谱(high performance liquid chromatography, HPLC)叠加图谱,以牛磺胆酸钠结合能力评价各样品的降脂活性,通过建立样品共有峰与降脂活性的谱效关系,筛选潜在的降脂活性成分;并应用计算机虚拟筛选技术预测靶蛋白,结合分子对接预测其作用靶点,初步研究其降脂机制。结果 17个油橄榄果提取物样品均具有不同程度降脂能力,峰P6、P10、P13、P14、P15、P17、P19和P20为潜在的降脂活性成分,其中P6和P13鉴定为羟基酪醇和断氧化马钱子苷,二者分别与多不饱和脂肪酸脂氧合酶ALOX15和钠/葡萄糖共转运蛋白1的残基存在氢键、疏水相互作用及静电相互作用,进而发挥结合抑制作用。结论 油橄榄果提取物中活性成分不仅能通过与牛磺胆酸钠结合,而且还能与靶蛋白结合发挥降脂作用,具有多成分、多靶点、多途径的特点,本研究结果可为后续研究提供理论依据。

Objective To explore the active substance basis of in vitro lipid-lowering activity of Olea europaea L.fruit extract,and preliminarily investigate the mechanism of lipid-lowering action of active ingredient.Methods High performance liquid chromatography(HPLC)fingerprints of 17 Olea europaea L.fruit extract samples were established.The lipid-lowering activity of each sample according to the binding ability of sodium taurocholic acid was evaluated.The spectral relationship between the common peaks and lipid-lowering activity was established to screen the potential lipid-lowering active ingredients.Computer virtual screening technology was used to predict the target protein,and molecular docking was used to predict the target of action,and the mechanism of lipid-lowering was preliminarily studied.Results The 17 samples all had different lipid-lowering abilities.The P6,P10,P13,P14,P15,P17,P19 and P20 were potential lipid-lowering active components,and P6 and P13 were identified as hydroxytyrosol and secoxyloganin.They had hydrogen bonding,hydrophobic interaction and electrostatic interaction with residues of polyunsaturated fatty acid lipoxygenase ALOX15 and sodium/glucose cotransporter 1,respectively,and thus exerted binding inhibition.Conclusion The active ingredient in Olea europaea L.fruit extract can not only bind with sodium taurocholate,but also bind with target protein to play a lipid-lowering effect,which has the characteristics of multi-component,multi-target and multi-pathway,the results of this study can provide theoretical basis for subsequent research.