Study the interactions between multiple flavonoids and bovine serum albumin by the developed equilibrium dialysis

作者全名："Tian, Yan; Li, Chunhu; Zeng, Fanyu; Yu, Chao; Xia, Zhining; Huang, Yike"

作者地址："[Tian, Yan; Li, Chunhu; Zeng, Fanyu; Xia, Zhining] Chongqing Univ, Sch Pharmaceut Sci, Chongqing 401331, Peoples R China; [Yu, Chao; Huang, Yike] Chongqing Med Univ, Coll Pharm, Chongqing 400016, Peoples R China"

通信作者："Xia, ZN (通讯作者)，Chongqing Univ, Sch Pharmaceut Sci, Chongqing 401331, Peoples R China.; Huang, YK (通讯作者)，Chongqing Med Univ, Coll Pharm, Chongqing 400016, Peoples R China."

来源：JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES

ESI学科分类：CHEMISTRY

WOS号：WOS:000884838300002

JCR分区：Q2

影响因子：3

年份：2022

卷号：1212

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：Equilibrium dialysis; Bovine serum albumin; Flavonoids; Multi-components; Binding constant

摘要："The therapeutic function of traditional Chinese medicine (TCM) is based on the combination effect of multiple active ingredients. However, the current pharmacological studies mainly focus on the protein binding of the single component from TCM, which is difficult to explain the overall therapeutic mechanism. Thus in this work the equilibrium dialysis method combined with high performance liquid chromatography (HPLC) and ultra -performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed to study the interactions between multi-components and protein. Firstly, the binding constants of seven different structural types of flavonoids with bovine serum albumin (BSA) were determined. The results showed that the binding affinity of flavones and flavonols with BSA was stronger than that of dihydroflavonoids, and the substitution of glycosides would reduce the binding affinity with BSA. The results of competitive displacement experiment showed that there existed competitive interactions among the four flavonoids (rutin, luteolin, hesperetin and kaempferol). The binding constants of flavonoids to BSA were significantly changed under the condition of multi -components coexistence. Especially, the binding constant of hesperetin to BSA increased from 9.46 x 104 L/mol to 1.49 x 106 L/mol under the coexistence of rutin. The results of fluorescence spectroscopy showed that the reason for competitive binding was that the four flavonoids were mainly bound to the IIA region of BSA. Finally, the method was successfully applied to study the binding of multiple components in Radix Scutellariae (RS) extract with BSA. Five flavonoids in RS extract were identified by UPLC-MS/MS, they had different degrees of binding to BSA, among which oroxylin A had the strongest binding degree. In conclusion, the equilibrium dialysis was reliable and sufficiently accurate for study of the interaction between multi-components or TCM extract and protein, which can provided a theoretical basis for the scientific explanation of the overall treatment mechanism of TCM."

基金机构："National Natural Science Foundation of China (NSFC) [21974015, 21705012]"

基金资助正文："This work was supported by the National Natural Science Foundation of China (NSFC, 21974015 and 21705012) ."