基于多光谱法的3种黄酮类化合物与胰脂肪酶的结合作用研究

Study on the binding effects of 3 kinds of flavonoids with pancreatic lipase based on multi-spectroscopy

中文关键词:  黄芩素  槲皮素  漆黄素  多光谱法

英文关键词:baicalein  quercetin  fisetin  multi-spectroscopy

中文摘要:

      目的 研究黄芩素(baicalein, Bai)、槲皮素(quercetin, Que)和漆黄素(fisetin, Fis) 3种黄酮类化合物与胰脂肪酶(pancreatic lipase, PL)的结合作用。方法 采用紫外-可见光谱法、三维荧光光谱法和傅里叶变换红外光谱法(Fourier transform infrared spectroscopy, FTIR)研究3种黄酮类化合物与PL的相互作用机制, 采用差示热量扫描法(differential scanning calorimetry, DSC)考查复合物中PL的稳定性, 计算配体的保留率。结果 紫外-可见吸收光谱表明3种黄酮类化合物与PL通过静态猝灭形成复合物, 结合常数分别为5.06×103、4.86×103和3.83×103 L/mol。三维荧光光谱表明, 3种黄酮类化合物诱导了PL的微环境和构象变化。FTIR表明Bai、Que和Fis改变了PL多肽骨架网络结构和氢键相互作用。同时复合物的形成稳定了PL的结构并提高了热稳定性, PL变性温度Tm从77.91℃升高至79.50、78.29、79.06℃。热稳定性表明PL增加3种黄酮化合物的保留率, 减少活性成分的损失。结论 Bai、Que和Fis能改变PL的构象变化, 并且复合物的形成提高了PL的稳定性。

英文摘要:

      Objective To investigate the binding effects of baicalein (Bai), quercetin (Que) and fisetin (Fis) with pancreatic lipase (PL). Methods The interaction mechanisms of the 3 kinds of flavonoids with PL were studied by ultraviolet-visible absorption spectroscopy, three-dimensional fluorescence spectroscopy and Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) was used to investigate the stability of PL in the complexes and calculate the retention rates of the ligands. Results The ultraviolet-visible absorption spectroscopy showed that the 3 kinds of flavonoids formed compounds with PL through static quenching, and the binding constants were 5.06×103, 4.86×103 and 3.83×103 L/mol, respectively. Three-dimensional fluorescence spectra showed that the microenvironment and conformational changes of PL were induced by the 3 kinds of flavonoids. FTIR showed that Bai, Que and Fis changed the skeleton network structure and hydrogen bond interaction of PL polypeptide. At the same time, the formation of complex stabilized the structure of PL and improved the thermal stability. The denaturation temperatures of PL increased from 77.91℃ to 79.50, 78.29, 79.06℃. Thermal stability showed that PL increased the retention rates of 3 kinds of flavonoid compounds and reduced the loss of active ingredients. Conclusion Bai, Que and Fis can change the conformational change of PL, and the formation of complex can improve the stability of PL.