摘要
目的:采用原代大鼠肝细胞(PRH)研究扶正化瘀方(FZHY)对大鼠细胞色素P450酶(CYP450)的影响。方法:采用两步灌流法分离大鼠原代肝细胞,形成肝板结构后加入FZHY共培养24 h,换成含有4种CYP450同工酶(CYP1A2,CYP2C9,CYP2D6,CYP3A4)对应的特异性探针药物(非那西丁、甲苯磺丁脲、氢溴酸右美沙芬、睾酮)的培养基,培养2 h后吸取上清液,利用UPLC-MS/MS对探针药物的代谢产物进行检测,考察FZHY对探针药物代谢产物生成量的影响,评价FZHY对PRH中CYP450代谢活力的影响。结果:PRH中CYP1A2,CYP2C9,CYP2D6与CYP3A4具有良好的代谢活力。4种探针药物在0-500μmol·L(-1)内对PRH没有毒性,在0.5-500μg·L(-1)时,FZHY对PRH没有毒性。FZHY在5μg·L(-1)时对PRH具有轻微的保护作用,并能使甲苯磺丁脲的代谢活化值——米氏常数(Km)降低20.8%;氢溴酸右美沙芬的Km降低39.2%;非那西丁的Km降低17.4%;并使睾酮的Km显著降低。结论:FZHY能抑制原代大鼠肝细胞中CYP1A2,CYP2C9,CYP2D6和CYP3A4的代谢活性。
Objective:Taking primary rat hepatocytes(PRH) model,to explore the effect of Fuzheng Huayu recipe(FZHY) on rat cytochrome P450 enzyme(CYP450).Method:PRH were isolated and cultured by two-step perfusion method.With the formation of the hepatic plate structure,FZHY was added to culture for 24 h,then replaced with culture medium containing specific probe substrates of CYP450(phenacetin-CYP1A2,tolbutamide-CYP2C9,dextromethorphan hydrobromide-CYP2D6,testosterone-CYP3A4),the supernatant was aspirated after 2 h,the corresponding products of the probe drugs were detected by UPLC-MS/MS.Result:CYP1A2,CYP2C9,CYP2D6 and CYP3A4 in PRH had good metabolic activity.The probe drugs had no toxicity on PRH in the range of 0-500 μmol·L^(-1),FZHY had no toxicity on PRH in the range of 0.5-500 μg·L(-1).FZHY could slightly protect PRH with concentration of 5 μg·L(-1).After PRH was cultured with 5 μg·L(-1) FZHY for 24 h,Michaelis constant(Km) of tolbutamide decreased by 20.8%,Kmof dextromethorphan hydrobromide decreased by39.2%,Kmof phenacetin decreased by 17.4%,Kmof testosterone decreased significantly.Conclusion:FZHY can inhibit the activity of CYP1A2,CYP2C9,CYP2D6 and CYP3A4 in PRH.
作者
张亚蕾
郑天慧
刘伟
陶艳艳
王长虹
杨涛
刘成海
ZHANG Ya-lei;ZHENG Tian-hui;LIU Wei;TAO Yan-yan;WANG Chang-hong;YANG Tao;LIU Cheng-hai(Institute of Liver Diseases, Institute of Cardiovascular Disease, Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shuguang Hospital Affiliated to Shanghai University of Traditmnal Chinese Medicine (TCM) , Shanghai 201203, China;School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China;Institute of Chinese Materia Medica, Shanghai University of TCM, Key Laboratory for Standardization of Chinese Medicines, Ministry of Education, Key Laboratory of Comprehensive Evaluation of New Resources and Quality Standards of TCM, State Adiministration of TCM, Shanghai 201203, China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2018年第9期22-28,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家国际科技合作专项(2014DFA31440)
上海市医学领军人才计划项目(LJ10005)
上海中医药事业发展三年行动计划项目(ZY3-CCCX-2-1003)
关键词
扶正化瘀方
细胞色素P450酶
原代肝细胞
中药复方
非那西丁
甲苯磺丁脲
氢溴酸右美沙芬
睾酮
Fuzheng Huayu recipe
cytochrome P450 enzyme
primary hepatocytes
compound Chinese medicine
phenacetin
tolbutamide
dextromethorphan hydrobromide
testosterone