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目的通过体内、外实验,观察盐酸双苯氟嗪对大鼠CYP450酶活性的影响。方法在正常大鼠肝微粒体温孵反应系统中加入不同浓度盐酸双苯氟嗪(0~200μmol·L~(-1))和探针药物共同温孵,LC-MS/MS测定各组探针药物代谢产物比值,观察盐酸双苯氟嗪对CYP450酶活性的影响。雄性SD大鼠随机分为空白组、盐酸双苯氟嗪低、中、高剂量组和苯巴比妥组,分别灌胃给予盐酸双苯氟嗪30、60、90 mg·kg~(-1)或苯巴比妥120 mg·kg~(-1),连续给药14 d。第15天灌胃给予Cocktail探针药物,于灌胃后不同时间眼内眦取血,LC-MS/MS测定血浆中探针药物浓度,绘制药-时曲线,计算药动学参数。取血结束后,制备肝微粒体。在肝微粒体温孵反应系统中加入探针药物,LC-MS/MS测定探针药物及其代谢物的浓度,计算探针药物的代谢率。通过比较实验组和空白对照组的相对肝脏重量、蛋白浓度、CYP450酶含量、药-时曲线、药动学参数和探针药物的代谢率,确定盐酸双苯氟嗪对CYP450酶活性的影响。结果在正常大鼠肝微粒体温孵液中,盐酸双苯氟嗪对CYP2C6、CYP2D1和CYP2C11有抑制作用,其IC50值分别是8.85、20.93和69.45μg·m L~(-1)。大鼠被不同剂量盐酸双苯氟嗪诱导14 d后,大鼠相对肝脏重量、蛋白浓度和CYP450酶含量没有影响;而苯巴比妥则使大鼠相对肝脏重量、蛋白浓度和CYP450酶含量显著增加。药动学参数和探针药物实验结果均显示,盐酸双苯氟嗪低剂量仅对CYP2C11有诱导作用;中剂量抑制CYP2D1,诱导CYP2C11和CYP3A;高剂量抑制CYP2C6和CYP2D1,诱导CYP2C11和CYP3A。苯巴比妥对CYP1A2、CYP2C11、CYP2D1、CYP3A均有诱导作用。结论正常大鼠肝微粒体温孵及整体动物实验均表明,盐酸双苯氟嗪可显著抑制CYP2C6和CYP2D1,但对CYP2C11则表现为体外抑制而体内诱导。
Objective To observe the effect of dipfluzine hydrochloride on the activity of CYP450 in rats by in vitro and in vivo experiments. Methods Different concentrations of dipfluzine hydrochloride (0 ~ 200μmol·L -1)) and probe drugs were incubated in the normal rat liver microsomal incubation system. LC-MS / MS was used to detect the concentration of probe Drug metabolite ratio observed dipyridafos hydrochloride CYP450 enzyme activity. Male Sprague-Dawley rats were randomly divided into blank group, low, medium and high doses of dipfluzine hydrochloride group and phenobarbital group, respectively, given diphenhydramine hydrochloride 30,60,90 mg · kg ~ (-1) ) Or phenobarbital 120 mg · kg -1 for 14 days. On the 15th day, the Cocktail probe was given intragastrically, and the blood was collected from the eye at different time after gavage. The concentration of the probe drug in the plasma was determined by LC-MS / MS, and the pharmacokinetic parameters were calculated. After the blood was taken, liver microsomes were prepared. In the liver microsomal incubation reaction system by adding probe drugs, LC-MS / MS probe drug and its metabolite concentration, calculate the probe drug metabolic rate. The effects of dipfluzine hydrochloride on CYP450 activity were determined by comparing the relative liver weight, protein concentration, CYP450 enzyme activity, drug-time curve, pharmacokinetic parameters and the metabolic rate of the probe drug in the experimental group and the blank control group. Results Dipfluzine hydrochloride had inhibitory effects on CYP2C6, CYP2D1 and CYP2C11 in normal rat liver microsomes with IC50 values of 8.85, 20.93 and 69.45μg · m L -1, respectively. Rats were induced by different doses of dipfluzine hydrochloride for 14 days, the relative liver weight, protein concentration and CYP450 enzyme content in rats had no effect; and phenobarbital rats relative liver weight, protein concentration and CYP450 enzyme content significantly increase. Pharmacokinetic parameters and probe drug results show that low doses of dipfluzine hydrochloride only CYP2C11 induction; middle dose CYP2D1, induced CYP2C11 and CYP3A; high dose CYP2C6 and CYP2D1 inhibition, induced CYP2C11 and CYP3A. Phenobarbital can induce CYP1A2, CYP2C11, CYP2D1 and CYP3A. Conclusion Normal rat liver microsome incubation and whole animal experiments show that dipfluzine hydrochloride can significantly inhibit CYP2C6 and CYP2D1, but CYP2C11 showed in vitro inhibition in vivo induction.