Cocktail探针药物评价淫羊藿提取物对大鼠CYP450酶的影响

Evaluation of Herba Epimedii extracts on CYP450 enzyme activity of SD rats by cocktail probe drugs

目的研究淫羊藿提取物对大鼠CYP450酶系的影响。方法将SD雄性大鼠随机分成2组(均n=20),用药组给予淫羊藿提取物(相当于淫羊藿总黄酮300mg·kg-1,ig,qd×14d),空白对照组给予生理盐水,于d15日清晨灌胃给予Cocktail探针药物咖啡因(15mg·kg-1)、氨苯砜(15mg·kg-1)、苯妥英(75mg·kg-1)和氯唑沙宗(30mg·kg-1)。采用RP-HPLC法检测各探针药物浓度,通过药动学参数来评价各组CYP450酶系的活性。结果咖啡因、氨苯砜、苯妥英和氯唑沙宗的线性范围分别为0.2～1.6mg·L-1、0.075～0.600mg·L-1、0.2～1.6mg·L-1、0.25～2.00mg·L-1,4种探针药物日内、日间RSD均<5.14%,方法回收率均>95.0%(RSD<6.14%)。给予淫羊藿提取物的大鼠CYP1A2、CYP3A4、CYP2C9和CYP2E1的活性水平显著高于对照组(P<0.05)。结论该RP-HPLC法快速、准确、灵敏度高、专属性强,可用于探针药物评价主要CYP450亚型酶的活性。淫羊藿提取物对大鼠CYP1A2、CYP3A4、CYP2C9和CYP2E1的活性均有诱导作用。

AIM To evaluate the effect of Herba Epimedii extracts on CYP450 enzyme activity by cocktail probe drugs in male SD rats. METHODS Male SD rats were randomly divided into two groups （each n = 20）. One group of the rats was given oeal Herba Epimedii extracts once daily for 14 d while the other group was received orally physiological saline also once daily as the blank control group. A series of cocktail probe drugs including caffeine （15mg·kg^-1）, dapsone （15mg·kg^-1）, phenytoin （75mg·kg^-1）, and chlorzoxazone （30 mg·kg^-1） were respectively infused into stomach of rats in the morning of d 15 as the probes of cytochrome P450 （CYP） enzyme isoform of CYP1A2, CYP3A4, CYP2C9, and CYP2E1 after multiple dosing of Herba Epimedii extracts. The plasma concentrations of the four probes were determined by RP-HPLC and their corresponding pharmacokinetic parameters were calculated to evaluate the possible CYP450 enzyme activity. RESULTS dapsone, phenytoin and chlorzoxazone The linear concentration ranges were 0.2 - 1.6 mg·L^-1, 0.075 effect of the of Herba Epimedii extracts on calibratio 0.600 mg·L^-1, n curves for caffeine, 0.2 - 1.6mg·L^-1 and 0.25 - 2.00 mg·L^-1, respectively. The intra-day and inter-day relative standard deviations （RSD） were all less than 5.14%. The methodological recoveries of the four probes were all more than 95.0% （RSD 〈 6.14%）. The activities of the CYP1A2, CYP3A4, CYP2C9 and CYP2E1 in the rats received Herba Epimedii extracts were significantly enhanced （P 〈 0.05）, in comparing with the blank control group. CONCLUSION The RP-HPLC method is rapid, sensitive, precise and specific, providing the major measure for determining the plasma concentration of these probes and evaluating the CYP1A2, CYP3A4, CYP2C9 and CYP2E1 of CYP450 subtype ebzymic activities in the rats with simultaneous induction for all.