小檗碱类成分对乙酰胆碱酯酶的抑制作用及血脑屏障透过性分析  

Analysis of Inhibitory Effect of Berberine Compounds on Acetylcholinesterase and Blood-brain Barrier Permeability

作　　者：潘福璐 刘洋[1] 刘慧宁 于爽 李雪岩 王新宇 祁东盈 柴晓钰 王倩倩 易紫容 潘艳丽[2] PAN Fulu;LIU Yang;LIU Huining;YU Shuang;LI Xueyan;WANG Xinyu;QI Dongying;CHAI Xiaoyu;WANG Qianqian;YI Zirong;PAN Yanli(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China;Institute of Information on Traditional Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)

机构地区：[1]北京中医药大学中药学院,北京102488 [2]中国中医科学院中医药信息研究所,北京100700

出　　处：《中国实验方剂学杂志》2024年第9期116-124,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973295);中国中医科学院科技创新工程项目(CI2021A00511)。

摘　　要：目的:从药靶结合亲和力和动力学角度研究非洲防己碱等小檗碱类成分对乙酰胆碱酯酶的抑制作用;并在不同多成分背景下研究这类成分的血脑屏障透过性。方法:采用Ellman改良的光度检测法测量非洲防己碱等小檗碱类成分对乙酰胆碱酯酶的半数抑制浓度(IC50);采用酶活恢复方法测定这类成分与乙酰胆碱酯酶结合的动力学参数(Koff);采用超高效液相色谱-高分辨率质谱法(UPLC-HRMS)对各成分在不同背景下(药材与复方)透过血脑屏障到达脑组织的定性分析。结果:非洲防己碱等小檗碱类成分对乙酰胆碱酯酶有较强的抑制活性,其IC_(50)可达到纳摩尔级,并且多个生物碱成分具有优于阳性药盐酸多奈哌齐的药靶结合动力学特征(更小的Koff,P<0.01),表明该类成分可在乙酰胆碱酯酶上持续更长时间的抑制;非洲防己碱等小檗碱类成分以单体形式、以及在黄连和黄柏药材提取物和复方黄连解毒汤的多成分背景下都可以透过血脑屏障到达脑组织。结论:非洲防己碱等小檗碱类成分对乙酰胆碱酯酶具有较强的抑制作用且在多种成分背景下均可入脑。这在药效物质基础层面上可以佐证临床上复方黄连解毒汤改善阿尔茨海默病的效果,为阐明复方黄连解毒汤的药效物质基础提供了数据支持。Objective:To examine the inhibitory effects of berberine compounds,including columbamine,on acetylcholinesterase from the perspectives of drug-target binding affinity and kinetics and explore the blood-brain barrier(BBB)permeability of these compounds in different multi-component backgrounds.M ethod:The median inhibitory concentration(IC50)of acetylcholinesterase by berberine compounds including columbamine was measured using the Ellman-modified spectrophotometric method.The binding kinetic parameters(Koff)of these compounds with acetylcholinesterase were determined using the enzyme activity recovery method.A qualitative analysis of the ability of these components to penetrate the BBB and arrive at the brain tissue in diverse multi-component backgrounds(including medicinal herbs and compound formulas)was conducted using ultra performance liquid chromatography-high resolution mass spectrometry(UPLC-HRMS).Result:Berberine compounds,including columbamine,exhibited strong inhibition of acetylcholinesterase,with IC_(50)values in the nanomolar range.Moreover,they displayed better drug-target binding kinetics characteristics(with smaller Koff values)than the positive control of donepezil hydrochloride(P<0.01),indicating a longer inhibition duration of acetylcholinesterase.Berberine components such as columbamine could penetrate the BBB to arrive at brain tissue in the form of a monomer,as well as in the multi-component backgrounds of Coptis and Phellodendri Chinensis Cortex medicinal extracts and the compound formula Huanglian Jiedutang.Conclusion:These berberine compounds such as columbamine exhibit a strong inhibitory effect on acetylcholinesterase and can arrive at brain tissue in multi-component backgrounds.In the level of pharmacological substance,this supports the clinical efficacy of compound Huanglian Jiedutang in improving Alzheimer's disease,providing data support for elucidating the pharmacological basis of compound Huanglian Jiedutang.

关 键 词：非洲防己碱 乙酰胆碱酯酶 药靶结合 亲和力 血脑屏障 

分 类 号：R284.2[医药卫生—中药学] R285[医药卫生—中医学] R289R287R22R2-031R33R24