徐焕华;汪美汐;谭洪玲;王宇光;汤响林;肖成荣;李桦;高月;马增春;
XU Huan-hua;WANG Mei-xi;TAN Hong-ling;WANG Yu-guang;TANG Xiang-lin;XIAO Cheng-rong;LI Hua;GAO Yue;MA Zeng-chun;Institute of Radiation Medicine,Academy of Military Medical Sciences;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences;Guangdong Pharmaceutical University;

• 1:军事医学科学院放射与辐射医学研究所
• 2:军事医学科学院毒物与药物研究所
• 3:广东药科大学

摘要(Abstract)：

探究临床用量的复方黄黛片及大剂量君药雄黄对大鼠肝脏主要药物代谢酶(CYP450)酶活性的影响及在mRNA水平的调控作用。以Wistar大鼠为研究对象,受试药物给予14 d后,采用Cocktail体外孵育法结合HPLC-MS/MS技术对大鼠肝中CYP1A2,CYP2B,CYP3A,CYP2C各亚酶的活性进行测定,并利用实时荧光定量PCR技术对上述亚型的mRNA水平表达进行检测。结果显示,复方黄黛片显著诱导CYP1A2,CYP2B酶活性,抑制CYP3A酶活性,结果与mRNA的表达具有一致性,但其单味药却呈现出弱于其甚至是相反的现象;不同剂量的雄黄组之间对酶活性及mRNA的表达结果亦呈现非常显著的不一致性。这些现象可能与复方黄黛片的配伍增效或减毒有关。CYP450酶的研究结果亦可以提示,复方黄黛片与其他药物合用时有可能产生药物相互作用。
To investigate the effect of clinical dose of Realgar-Indigo Naturais formula( RIF) and large-dose of Realgar on main drug-metabolizing enzymes CYP450 s of rat liver,as well as its regulatory effect on mRNA expression. Wistar rats were administrated orally with tested drugs for 14 days. A Cocktail method combined with HPLC-MS/MS was used in the determination of 4 cytochrome P450 isozymes( CYP1A2,CYP2 B,CYP3A and CYP2C) in liver of the rats,and the mRNA expression levels of the above subtypes were detected by real-time fluorescent quantitative PCR. The results showed that RIF can significantly induce CYP1A2 and CYP2 B enzyme activity,and inhibit CYP3 A enzyme activity. This result was consistent with the mRNA expression. However,its single compound showed weaker or even contrary phenomenon. Different doses of Realgar also showed significant inconsistencies on CYP450 enzymes activity and mRNA expression. These phenomena may be relevant with RIF compatibility synergies or toxicity reduction. The results can also prompt drug interactions when RIF is combined with other medicines in application.

关键词(KeyWords)： 复方黄黛片;雄黄;细胞色素P450;肝微粒体;酶活性
Realgar-Indigo Naturalis formula;realgar;cytochrome P450;liver microsome;enzyme activity

Abstract:

Keywords:

基金项目(Foundation): 国家“重大新药创制”科技重大专项(2015ZX09501004-003-003)

作者(Authors): 徐焕华;汪美汐;谭洪玲;王宇光;汤响林;肖成荣;李桦;高月;马增春;
XU Huan-hua;WANG Mei-xi;TAN Hong-ling;WANG Yu-guang;TANG Xiang-lin;XIAO Cheng-rong;LI Hua;GAO Yue;MA Zeng-chun;Institute of Radiation Medicine,Academy of Military Medical Sciences;Institute of Pharmacology and Toxicology,Academy of Military Medical Sciences;Guangdong Pharmaceutical University;

DOI: 10.19540/j.cnki.cjcmm.20170103.019

参考文献(References)：

• [1]Chen X M,Wei M,Zhang H M,et al.Effect of vanillin and ethyl vanillin on cytochrome P450 activity in vitro and in vivo[J].Food Chem Toxicol,2012,50(6):1897.
• [2]Han Y L,Li D,Ren B,et al.Evaluation of impact of Herba Erigerontis injection,a Chinese herbal prescription,on rat hepatic cytochrome P450 enzymes by cocktail probe drugs[J].J Ethnopharmacol,2012,139:104.
• [3]Badal S,Williams S A,Huang G,et al.Cytochrome P450 1 enzyme inhibition and anticancer potential of chromene amides from Amyris plumieri[J].Fitoterapia,2011,82(2):230.
• [4]童瑞敏.何首乌及其制剂致药物性肝损伤20例分析[J].山东中医杂志,2015,34(12):928.
• [5]Wang Y G,Zhou J M,Ma Z C,et al.Pregnane X receptor mediated-transcription regulation of CYP3A by glycyrrhizin:a possible mechanism for its hepatoprotective property against lithocholic acid-induced injury[J].Chem Biol Interact,2012,200(1):11.
• [6]赵春,李银生,蒋美琳,等.细胞色素P450与外源物的相互作用研究进展[J].中国现代应用药学,2014,31(8):1020.
• [7]高月.基于药物代谢酶的中药毒性与毒理研究[J].毒理学杂志,2005,19(3):175.
• [8]Torka P,Al Ustwani O,Wetzler M,et al.Swallowing a bitter pill-oral arsenic trioxide for acute promyelocytic leukemia[J].Blood Rev,2016,30(3):201.
• [9]孙峰,陈楠楠,成玉斌.复方黄黛片治疗急性早幼粒细胞白血病204例经验总结[J].中西医结合学报,2008,6(6):639.
• [10]许龙龙,汤响林,马增春,等.决明子水提液对大鼠肝脏CYP450酶的影响[J].中国中药杂志,2016,41(8):1504.
• [11]梁淼.基于P450酶的四物汤配伍规律研究[D].南宁:广西医科大学,2014.
• [12]韦惠琴,钟卫一.复方黄黛片联合全反式维甲酸治疗初发急性早幼粒细胞白血病的疗效[J].中国医药导刊,2013,15(11):1864.
• [13]谢玮,庞缨,叶絮,等.复方黄黛片联合沙利度胺治疗难治或复发多发性骨髓瘤临床观察[J].广州医科大学学报,2015,43(5):28.
• [14]Ingelman-Sundberg M.Pharmacogenetics of cytochrome P450 and its applications in drug therapy:the past,present and future[J].Trends Pharmacol Sci,2004,25(19):193.
• [15]Lewis D F.P450 structures and oxidative metabolism of xenobiotics[J].Pharmacogenomics,2003,4(4):387.
• [16]梁爱华,李春英,王金华,等.雄黄的毒性研究[J].中国中药杂志,2011,36(12):1889.
• [17]陆远富,时京珍,石京山,等.科学评价含雄黄、朱砂中成药的安全性[J].中国中药杂志,2011,36(24):3402.
• [18]徐芝秀,石秀英,金科涛,等.甘草与海藻提取液合用对CYP3A1/2酶活性及mRNA表达的影响[J].中国药师,2007,10(6):515.