临床用量的复方黄黛片及大剂量雄黄对大鼠肝脏主要药物代谢酶的影响

Effect of clinical doses of Realgar-Indigo Naturalis formula and large-dose of realgar on CYP450s of rat liver

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.