汤阴北艾精油对鼠CYP450酶活性和表达的影响

doi:10.16431/j.cnki.1671-7236.2019.11.035

摘要/Abstract

摘要： 试验旨在研究汤阴北艾精油对CYP450酶各亚型活性和表达的影响。采用水蒸馏提取法提取北艾精油，通过GC-MS检测北艾精油成分。为研究北艾精油对CYP450酶活性的影响，将SD大鼠随机分为对照组（CON）和北艾精油组（EO），分别灌胃0.1%吐温-80 100 μL/g和0.1%吐温-80稀释的北艾精油（5%）100 μg/g，灌胃后给予20 μg/g各探针药物，采用鸡尾酒探针法分别对给药后5、10、20、30 min及1、2、3、6、12、24、36、48 h血浆中3种探针底物的浓度进行检测，考察北艾精油对CYP1A2、CYP2E1和CYP2D6酶活性的影响；为探究北艾精油对CYP450酶基因和蛋白表达的影响，将昆明小鼠随机分为对照组和北艾精油组，分别灌胃0.1%吐温-80 100 μL/g和0.1%吐温-80稀释的北艾精油（5%）100 μg/g，连续灌胃3日，每日2次，末次给药24 h后提取总mRNA及肝微粒体，采用实时荧光定量PCR和蛋白免疫印迹法（Western blotting）测定CYP1A2、CYP2E1和CYP2D6 mRNA和蛋白表达含量。结果表明：①北艾精油主要成分为Eucalyptol；3-Cyclohexen-1-ol，4-methyl-1-（1-methylethyl）-3；（+）-2-Carene，4-.alpha.-isopropenyl-2；m-Mentha-4，8-diene，（1S，3S）-（+）-；Benzoic acid，2，4，6-trimethyl-2，4，6-trimethylphenyl ester。②对血液中非那西汀（CYP1A2底物）、右美沙芬（CYP2D6底物）和氯唑沙宗（CYP2E1底物）进行药代动力学分析可知，北艾精油具有抑制CYP2D6和CYP2E1酶活性的作用，而对CYP1A2酶活性无显著作用。③实时荧光定量PCR和Western blotting测定结果表明，北艾精油可显著抑制CYP2E1和CYP2D6酶的表达。综上，北艾精油对CYP2E1和CYP2D6酶活性和表达均具有明显的抑制作用，提示艾草精油不能同时与以CYP2D6和CYP2E1为主要代谢酶的药物同食。该研究结果可为北艾精油的合理用药提供数据基础。

关键词: 艾草; 精油; 药代动力学; CYP450酶

Abstract: Effect of essential oil from Artemisia vulgaris on CYP450 isoforms activity and expression in mouse was detected in this study.Essential oil was extracted from Artemisia vulgaris by steam extraction and was detected by GC-MS.In order to study the effect of essential oil on CYP450 activity,SD rats were randomly divided into control group (CON) and essential oil group (EO).The rats were given 0.1% Tween-80 100 μL/g and 0.1% Tween-80 diluted essential oil (5%) 100 μg/g respectively by gavage.After gavage,20 μg/g probes were given.Cocktail probes were used to measure the concentrations of three probe at 5,10,20 and 30 min,and 1,2,3,6,12,24,36 and 48 h after administration.To study the essential oil on the expression of CYP450,the Kunming mice were randomly divided into two groups.The control group was administrated with 1% Tween-80 (100 μL/g) and the essential oil group with 5% essential oil (100 μg/g).The administration was given twice daily for 3 days.The mRNA and protein expression were tested by Real-time PCR and Western blotting.The results show that:①The major components of essential oil were Eucalyptol;3-Cyclohexen-1-ol,4-methyl-1-(1-methylethyl)-3;(+)-2-Carene,4-.alpha.-isopropenyl-2;m-Mentha-4,8-diene,(1S,3S)-(+)-;Benzoic acid,2,4,6-trimethyl-,2,4,6-trimethylphenyl ester.②Pharmacokinetics of Phenacetin(CYP1A2 substrate),dextromethorphan(CYP2D6 substrate) and chlorzoxazone(CYP2E1 substrate) were analyzed.The results suggested that the essential oil could inhibit the activities of CYP2D6 and CYP2E1,while,had no effect on the activity of CYP1A2.③Real-time PCR and Western blotting results showed that essential oil significantly decreased the CYP2E1 and CYP2D6 expression.In conlusion,essential oil from Artemisia vulgaris could inhibit the activities and expressions of CYP2E1 and CYP2D6.It suggested that the essential oil from Artemisia vulgaris could not be taken together with the drugs in which metabolic pathway CYP2D6 and CYP2E1 were the main factors.The results of this study could provide data basis for the rational use of essential oil from Artemisia vulgaris.

Key words: Artemisia vulgaris; essential oil; pharmacokinetics; CYP450

中图分类号:

S853.74

蒋志惠, 张静苗, 曲毅程, 张守泉, 常雪梅, 郭晓. 汤阴北艾精油对鼠CYP450酶活性和表达的影响[J]. 中国畜牧兽医, 2019, 46(11): 3440-3448.

JIANG Zhihui, ZHANG Jingmiao, QU Yicheng, ZHANG Shouquan, CHANG Xuemei, GUO Xiao. Effect of Essential Oil from Artemisia vulgaris on CYP450 Isoforms Activity and Expression[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3440-3448.

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