高表达β_1肾上腺素受体细胞膜色谱法靶向筛选夏天无中活性成分  

Targeted Screening of Active Ingredients from Corydlis Decumbentis Rhizoma Based on High Expression β_1 Adrenergic Receptor / Cell Membrane Chromatography Method

作　　者：王新[1] 薛晖[1] 岳媛[2] 徐丹丹[1] 彭小丽[1] 

机构地区：[1]山西医科大学药学院,太原030001 [2]清华大学生命科学院,北京100083

出　　处：《中国实验方剂学杂志》2015年第18期65-68,共4页Chinese Journal of Experimental Traditional Medical Formulae

摘　　要：目的:从夏天无中筛选与高表达β1肾上腺素受体(β1-AR)作用的活性成分,并考察其对β1-AR的抑制作用。方法:建立β1-AR细胞膜色谱与UPLC/MS联用分析方法筛选鉴别夏天无中的活性成分;用美托洛尔(10μmol·L-1),药根碱(1,5μmol·L-1)处理β1-AR/CHO-S 24 h,换异丙肾上腺素(50μmol·L-1)损伤细胞10 h后,ELISA法测定活性成分对β1-AR下游因子c AMP,PKA水平的影响,评价其对β1-AR的抑制作用。结果:药根碱是夏天无中作用于β1-AR的活性化合物;细胞实验结果表明,与ISO组相比,药根碱和美托洛尔对c AMP,PKA水平均有下调作用(P<0.01)。结论:药根碱是夏天无中具有β1-AR抑制作用的活性化合物;建立的β1-AR/CMC-offline-HPLC/MS分析方法可快速有效筛选出作用靶点明确的活性成分,且筛选结果与生物效应相一致。Objective： To screen out the active components from Corydlis Decumbentis Rhizoma （CDR） and investigate their inhibitory effect on β1-AR. Method： A high expression β1 adrenergic receptor/cell membrane chromatography-offline-uhra-performance liquid chromatography/mass spectrometry method （β1-AR/ CMC-Offline-UPLC/MS method） was used to scan and identify the effective components from extracts of CDR. β1- AR/CHO-S cells were cultured in metoprolol （10 μmol·L^-1） and active components of jatrorrhizine （JAT） （1, 5 μmol ·L^-1） for 24 h. Then, the cells were injured in DMEM containing ISO （50 μmol·L^-1） for 10 h. the effect of the active components on β1-AR downstream factors cAMP and PKA was determined by ELISA kits to evaluate their inhibitory effect on β1-AR. Result ： JAT was the active component of CDR and the experiment in vivo showed that in JAT and MT groups, concentrations of cAMP and PKA were lower than that in the ISO group （P 〈 0.01 ）. Conclusion： JAT was successfully isolated and identified by the β1-AR/CMC-offline-UPLC/MS system from CDR and had an inhibitory effect on β1-AR. The β1-AR/CMC-Offline-UPLC/MS method can be used to screen out active components with clear action targets in a quick and effective manner. The screening results were found to be significantly consistent with its pharmacological effect.

关 键 词：Β1肾上腺素受体 细胞膜色谱法 活性成分 药根碱 

分 类 号：R284.1[医药卫生—中药学]