Cocktail探针法评价牛樟芝醇提物对大鼠肝CYP450酶活性的作用  

Evaluation of the effect of ethanol extracts of antrodia cinnamomea on rat hepatic CYP450 enzyme activity by Cocktail Probe Method

作　　者：魏文增 王宫 Wei Wenzeng;Wang Gong(Fujian Academy of Chinese Medical Sciences,Fuzhou Fujian 350003)

机构地区：[1]福建省中医药科学院,福建福州350003

出　　处：《山西中医药大学学报》2024年第8期857-864,共8页Journal of Shanxi University of Chinese Medicine

基　　金：省属公益类科研院所基本科研专项(2021R1003005、2020R1003005);福建省中医药健康科技经济融合服务平台。

摘　　要：目的:研究牛樟芝醇提物对大鼠肝微粒体CYP450亚酶活性的影响。方法:建立大鼠肝微粒体体外代谢的HPLC法,采用Cocktail探针法评价牛樟芝醇提物对大鼠肝微粒体CYP450亚酶的作用。(1)分别精密称取美托洛尔、奥美拉唑、非那西汀、氯唑沙宗、甲苯磺丁脲、硝苯吡啶各适量,配制Cocktail混合探针底物溶液。(2)设活性组、无活性组和抑制剂组,活性组:大鼠肝微粒体活性未进行灭活处理;无活性组:大鼠肝微粒体被灭活,其他同活性组;抑制剂组(分别为奎宁丁、噻氯匹定、α-萘黄酮、二乙氨基二硫代甲酸钠、磺胺苯吡唑、酮康唑组):在活性组的基础上加抑制剂各2.5μl。检测各组CYP2D6、CYP2C19、CYP1A2、CYP2E1、CYP2C9、CYP3A4/5亚酶的相对活性。结果:与无活性组比较,活性组中CYP2D6、CYP2C19、CYP1A2、CYP2E1、CYP2C9、CYP3A4/5相对酶活性升高,差异均有统计学意义(P<0.05)。与活性组比较,牛樟芝高、中、低剂量组中CYP1A2和CYP3A4/5相对酶活性降低,差异有统计学意义(P<0.05);CYP2C9相对酶活性降低,但差异无统计学意义(P>0.05);高、中剂量组中CYP2D6相对酶活性降低,高剂量组中CYP2E1、低剂量组中CYP2C19相对酶活性降低,高剂量组中CYP2C19相对酶活性升高,差异均有统计学意义(P<0.05)。结论:牛樟芝醇提物对大鼠肝微粒体CYP450亚酶中CYP1A2、CYP3A4/5、CYP2C9、CYP2D6、CYP2E1的代谢活性在一定浓度范围内有抑制作用,而对CYP2C19亚酶的代谢活性在一定浓度范围内有促进作用。Objective:To investigate the effect of the ethanol extract of antrodia cinnamomea on the activity of rat hepatic microsomal CYP450 subenzymes.Methods:An in vitro metabolic HPLC method was established for rat hepatic microsomes,and the Cocktail probe method was used to evaluate the effect of the ethanol extract of Antrodia cinnamomea on rat hepatic microsomal CYP450 subenzymes.(1)Accurately weighed amounts of metoprolol,omeprazole,phenacetin,chlorzoxazone,tolbutamide,and nifedipine were used to prepare a Cocktail mixed probe substrate solution(.2)Active,inactive,and inhibitor groups were set up.In the active group,rat hepatic microsomes were not inactivated.In the inactive group,rat hepatic microsomes were inactivated,with other steps being the same as in the active group.In the inhibitor groups(quinidine,ticlopidine,α-naphthoflavone,disulfiram,sulfaphenazole,and ketoconazole groups,respectively),inhibitors(2.5μl each)were added to the active group.The relative activities of CYP2D6,CYP2C19,CYP1A2,CYP2E1,CYP2C9,and CYP3A4/5 subenzymes were detected.Results:Compared with the inactive group,the relative enzyme activities of CYP2D6,CYP2C19,CYP1A2,CYP2E1,CYP2C9,and CYP3A4/5 were elevated in the active group,with statistically significant differences(P<0.05).Compared with the active group,the relative enzyme activities of CYP1A2 and CYP3A4/5 were reduced in the high,medium,and low dose groups of antrodia cinnamomea,with statistically significant differences(P<0.05).The relative enzyme activity of CYP2C9 was reduced but without statistical significance(P>0.05).The relative enzyme activity of CYP2D6 was reduced in the high and medium dose groups,and the relative enzyme activity of CYP2E1 in the high dose group and that of CYP2C19 in the low dose group were reduced.The relative enzyme activity of CYP2C19 was elevated in the high dose group,all with statistically significant differences(P<0.05).Conclusion:The ethanol extract of antrodia cinnamomea has inhibitory effects on the metabolic activities of CYP1A2,CYP3A4/5,CYP2C

关 键 词：COCKTAIL 探针底物 HPLC 牛樟芝 大鼠肝微粒体 

分 类 号：R285.5[医药卫生—中药学]