摘要
目的采用亲和萃取法筛选延胡索中作用于β2-肾上腺素受体(β2-adrenoceptor,β2-AR)的活性成分。方法采用重氮盐法将β2-AR固载于大孔硅胶表面制备亲和萃取材料,工具药吸附等温线拟合法表征亲和材料生物活性,利用该材料对延胡索水提液中的活性成分进行筛选,离心法分离亲和材料和提取液中与受体未结合的成分,高效液相色谱-电喷雾-四极杆-飞行时间串联质谱法鉴定活性成分。结果延胡索水提液中与β2-AR结合的活性成分主要为生物碱类,分别为四氢非洲防己碱,紫堇碱,四氢小檗碱和延胡索乙素。结论与受体色谱法相比,β2-AR亲和萃取法在保持固定化受体活性方面具有一定优势,避免了高压装填色谱柱造成的受体活性损失,且在中药活性成分筛选方面具有快速、简便和高效的特点,为中药活性成分高通量筛选提供了参考。
Objective To screen the active compounds targeted to beta2-adrenoceptor(β2-AR) in the water extract from Corydalis yanhusuo. Methods β2-AR was immobilized onto macroporous silica gel by diazonium reaction to prepare the affinity extraction material. The material was characterized by plotting the adsorption isotherm of the specific drugs and the gel. Water extract of Corydalis yanhusuo was screened by the affinity extraction material. The bound and unbound components to β2-AR were separated by centrifugation and the bound component was considered to be the bioactive compounds in Corydalis yanhusuo that target β2-AR. The bioactive compounds were identified by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry(HPLC-ESIQ-TOF-MS/MS). Results The main bioactive compounds in Corydalis yanhusuo were alkaloids, including tetrahydrocolumbamine, corydaline, tetrahydroberberberine and tetrahydropalmatine. Conclusion Compared with receptor chromatography, the β2-AR affinity extraction method has the advantage of remaining the bioactivity of receptors. Because this method avoids the high pressure packing procedures in preparing the receptor chromatographic column. In addition, this method is fast, simple and has high efficiency in screening bioactive compounds in traditional Chinese medicine, thus provides a good reference for high throughput drug screening.
作者
骆晶
孟雪
胡卫涛
高秋洁
张选国
李警卓
Luo Jing;Meng Xue;Hu Weitao;Gao Qiujie;Zhang Xuanguo;Li Jingzhuo(Shaanxi Provincial Hospital of Traditional Chinese Medicine,Shaanxi 710003,China;Institute of Traditional Chinese Medicine of Shaanxi Province,Shaanxi 710003,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2020年第10期3768-3774,共7页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
陕西省科学技术厅重点研发计划一般项目(2019SF-280):远志汤防治阿尔兹海默病作用机制及体内效应物质辨识技术研究,负责人:孟雪。
关键词
亲和萃取
β2-肾上腺素受体
延胡索
活性成分
筛选
Affinity extraction
β2-adrenoceptor
Corydalis yanhusuo
Bioactive compounds screening
作者简介
通讯作者:李警卓,主治医师,主要研究方向:中医药防治疾病的理论和临床研究。
