基于毛细管电泳的苦参碱转运过程中的血清白蛋白行为成像

引用本文: 赵富荣, 郭明, 邵东伟, 夏琪涵. 基于毛细管电泳的苦参碱转运过程中的血清白蛋白行为成像[J]. 色谱, 2020, 38(8): 975-983. doi: 10.3724/SP.J.1123.2019.12034 shu

Citation: ZHAO Furong, GUO Ming, SHAO Dongwei, XIA Qihan. Behavioral imaging of serum albumin during matrine transport based on capillary electrophoresis[J]. Chinese Journal of Chromatography, 2020, 38(8): 975-983. doi: 10.3724/SP.J.1123.2019.12034 shu

基于毛细管电泳的苦参碱转运过程中的血清白蛋白行为成像

赵富荣 ^1, ,
郭明 ^{1,2,
,} ,
邵东伟 ^3,4, ,
夏琪涵 ^1,

1.
浙江农林大学林业与生物技术学院, 浙江杭州 311300;
2.
浙江农林大学理学院, 浙江杭州 311300;
3.
佳木斯大学药学院, 黑龙江佳木斯 154007;
4.
美国北德克萨斯大学机械与动力工程学院, 德克萨斯州丹顿 76201

通讯作者: 郭明, E-mail:guoming@zafu.edu.cn

基金项目:
the Basic Public Welfare Research Project of Zhejiang Province of China (No. LGN20B070001).

摘要: 利用毛细管电泳（CE）技术在体外条件下建立了苦参碱（MT）与血清白蛋白（SA）相互作用的分析方法。生理条件下通过淌度移动法和前沿分析法（FA）研究苦参碱与血清白蛋白的结合状况，构建配体（MT）-受体（SA）相互作用模型。其中，淌度移动法采用非线性拟合方法获得苦参碱与人血清白蛋白（HSA）结合参数；FA运用非线性回归方程、Scatchard方程、Klotz方程3种方程获得苦参碱与人血清白蛋白结合参数，分析其相互作用状况进而分析模型适用度。利用淌度移动法可知，人血清白蛋白与苦参碱表观结合常数K_B为8.072×10³ mol/L；利用FA法可知，采用非线性回归方程、Scatchard方程、Klotz方程3种方程获得苦参碱与人血清白蛋白表观结合常数K_B分别为1.434×10³、1.781×10³和2.133×10³ mol/L，且二者结合位点数在1.0左右，说明苦参碱与人血清白蛋白作用只有单一类型的结合位点。通过计算分析得到3个方程的相关系数（r），关系为r_Klotz > r_非线性 > r_Scatchard。结果表明：淌度移动法和FA法均适用于分析MT-HSA体系的结合状况；由于FA法可以计算出表观结合常数的同时又能计算出结合位点数，因而更适合MT-HSA体系的分析研究；分析比较得出3种方程之中Klotz方程为最适理论模型。结合参数表明，MT-HSA相互作用体系之间发生的结合反应为单一类型的结合位点且结合稳定。相关工作阐明了典型生物碱与血清白蛋白的相互作用机制，可为生物碱的血液输运机制的深入研究提供有益参考。

关键词:

English

Behavioral imaging of serum albumin during matrine transport based on capillary electrophoresis

1.
College of Forestry and Bio-technology, Zhejiang A & F University, Hangzhou 311300, China;
2.
College of Science, Zhejiang A & F University, Hangzhou 311300, China;
3.
School of Pharmacy, Jiamusi University, Jiamusi 154007, China;
4.
Department of Mechanical and Power Engineering, University of North Denton, Denton 76201, USA

Corresponding author: GUO Ming, guoming@zafu.edu.cn

Received Date: 02 January 2020
Fund Project:
the Basic Public Welfare Research Project of Zhejiang Province of China (No. LGN20B070001).

Abstract: Matrine (MT) is an alkaloid widely used in the treatment of tumor diseases. It is the main medicinal ingredient in the dried roots of kuh-seng (Sophora flavescens Ait). However, there have been few studies on its transport mechanism. Serum albumin (SA) is the most abundant protein in blood. SA combines easily with many substances, including MT. MT and human serum albumin (HSA) were analyzed by capillary electrophoresis (CE) under in vitro conditions. The capillary tubing was 50 μm. The total length of the capillary was 60 cm, the total effective length was 50 cm. The interaction models of ligand-receptor binding were constructed by the mobility and frontal analysis (FA) methods. The purpose of establishing the interaction model was to study the binding of MT and SA. The phosphate buffer solution (PBS, 0.02 mol/L) was prepared in double distilled water. All solutions were prepared in PBS (0.02 mol/L). All solutions were filtered twice through a 0.45 μm microporous membrane, degassed for 5 min at a time. In the mobility method, different gradient MT solutions were used as running buffers. Their concentrations were 1.0×10^-4-1.0×10^-3 mol/L, with the gradient of 1.0×10^-4 mol/L. And the HSA solution containing (0.5% (v/v)) acetone was used as test sample. Its concentration was 1.0×10^-5 mol/L. The nonlinear fitting method was used to obtain the binding parameters of MT and HSA. In the FA method, different gradient MT-HSA solutions were used as test samples. Their concentrations were 1.0×10^-4-1.0×10^-3 mol/L, with the gradient of 1.0×10^-4 mol/L. And the PBS solution (0.02 mol/L) was used as running buffer. Then three equations were used to obtain the binding parameters of MT and HSA. And the applicability of the models was analyzed using the binding parameters. These three equations were nonlinear regression equation, Scatchard linear equation, and Klotz linear equation. Using the mobility method, the apparent binding constant K_B was 8.072×10³ mol/L. According to the FA method, three apparent binding constants were obtained for MT and HSA. The apparent binding constant K_B of HSA and MT by nonlinear regression equation, Scatchard linear equation and Klotz linear equation were 1.434×10³, 1.781×10³ and 2.133×10³ mol/L. The comparison was as follows, K_{B(nonlinear regression equation)} < K_{B(Scatchard linear equation)} < K_{B(Klotz linear equation)}. The number of binding sites was about 1.0. It was indicating that MT had only a single type of binding site with HSA. By analyzing the applicability of the model, the correlation coefficients (r) of the three equations were obtained. The comparison was as follows, r_{(Klotz linear equations)} > r_{(nonlinear regression equations)} > r_{(Scatchard linear equations)}. The results showed that both the methods were all suitable for analyzing the MT-SA system. The FA method could calculate the apparent binding constants and the numbers of binding sites. Therefore, it was more suitable for the analysis of MT and HSA. And the Klotz linear equation was the best fit for the theoretical model among the three equations. The combined parameters indicated that the interaction of MT with HSA had only one binding site. And the binding of MT with HSA was stable. This experimental method could be used to determine the binding status of MT and HSA. It is useful to further explore the binding mechanism of MT and HSA. This work provides valuable information on the interaction mechanism of typical alkaloids with SA. It will be useful in studies of the blood transport mechanisms of alkaloids.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

基于毛细管电泳的苦参碱转运过程中的血清白蛋白行为成像

通讯作者: 郭明, E-mail:guoming@zafu.edu.cn

English

Behavioral imaging of serum albumin during matrine transport based on capillary electrophoresis

Corresponding author: GUO Ming, guoming@zafu.edu.cn

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

基于毛细管电泳的苦参碱转运过程中的血清白蛋白行为成像

通讯作者: 郭明, E-mail:guoming@zafu.edu.cn

English

Behavioral imaging of serum albumin during matrine transport based on capillary electrophoresis

Corresponding author: GUO Ming, guoming@zafu.edu.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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