目的 探讨注射用丹参总酚酸(冻干)(SLI)对人CYP450酶和P-糖蛋白体外抑制作用以及对大鼠CYP1A2和CYP3A体内诱导作用.方法 ①应用P450-GloTM CYP450检测试剂盒,通过化学发光法测定SLI和经典抑制剂对细胞色素P4501 A2(CYP1A2),CYP2 D6,CYP3A4,CYP2 C19和CYP2C9的IC50值,通过比较SLI和经典抑制剂对相应细胞色素P450亚型的IC50值来判断SLI对人CYP450酶的体外抑制作用.②Wistar大鼠分别iv给予SLI 3,10和30 mg· kg-1和诱导剂苯巴比妥钠20 mg· kg-1,采用探针底物法,通过比较代谢产物的生成速率来评价SLI对大鼠CYP1 A2和CYP3A的诱导作用.③应用ATP酶检测试剂盒,通过化学发光法测定ATP酶活性来评价SLI是否为P-gp的底物或抑制剂.结果 ①CYP1 A2,CYP2 C9,CYP2 C19,CYP2 D6和CYP3A4抑制剂的IC50与SLI对其的IC50进行比较(CYP1A2:0.12 μm0l·L-1vs840 μm ol·L-1;CYP2C9:3.362 μmol·L-1vs704 μmol·L-1;CYP2C19:3.236 μmol·L-1vs306 μm ol·L-1;CYP2D6:0.117 μm ol·L-1 vs2660 μm0l·L-1;CYP3A4:0.078 μmol·L-1 vs 1780 μmol·L-1).②与空白对照组(86.4±6.3)nmol· g-1· min-1相比,SLI3,10和30 mg· kg-1组CYP1 A2活性分别为83.4±6.6,82.5 ±4.0和(83.4±6.6)nmol ·g-1· min-1.与空白对照组(16.1±0.9)nmol·g-1· min-1比较,SLI3,10和30 mg· kg-1组CYP3A活性分别为15.7±0.6,15.9±0.7和(15.9 ±1.0)nmol·g-1 ·min-1,无显著性差异.③以临床血药浓度为依据设计的一系列浓度的SLI 0.0002,0.0006,0.002,0.006,0.017,0.052,0.156和0.468 g·L-1的ATP酶活性分别与空白对照组进行比较(5.8,5.3,5.8,5.5,5.8,5.2,,5.8,5.3,vs 5.75 μmol·g-1 ·min-1),无显著性差异.结论 SLI临床给药剂量既不能体外抑制人CYP1A2,CYP2 D6,CYP3A4,CYP2 C19和CYP2C9酶活性,也不能诱导大鼠CYP1 A2和CYP3A,同时也不是P-gp的体外抑制剂或底物.%OBJECTIVE To investigate the inhibitory effect of salvianolate lyophilized injection (SLI) on human cytochrome P450 (CYP450) and P-glycoprotein (P-gp) system in vitro and the inductive effect on rat CYP1A2 and CYP3A in vivo.METHODS ① IC50 of cytochrome P-4501A2 (CYP1A2.),CYP2Cg,CYP2C19,CYP2D6 and CYP3A4 inhibitors was evaluated by chemiluminescence using P450-GloTM Screening Systems.The inhibition on CYP450 in vitro from SLI was evaluated by IC50.② Wistar rats were sc given SLI 3,10 and 30 mg·kg-1 or inducer.The method of probe substrate was used for evaluating the induction of rat CYP1A2 and CYP3A from SLI by comparing the production rate of metabolites.③ The ATPase activities of SLI were determined by ATPase assay kit,which can evaluate whether SLI is a substrate or inhibitor of P-gp.RESULTS ① IC50 of inhibitors and SLI was compared (CYP1A2,0.012 μmol·L-1 vs840 μmol·L-1 ; CYP2Cg,3.362 μmol·L-1 vs704 μmol·L-1 ; CYP2C19,3.236 μmol·L-1 vs 306 μmol·L-1 ; CYP2D6,0.117 μmol·L-1 vs 2660 μmol·L-1 ; and CYP3A4,0.078 μmol·L-1 vs 1780 μmol·L-1).② Compared with control group (86.4 ±6.3)nrnol·g-1 ·min-1,the activities of rat CYP1A2 in SLI 3,10 and 30 mg·kg-1 groups were 83.3 ±6.6,82.5 ±4.0 and (83.4 ±6.6)μmol·g-1 ·min-1,respectively.Compared with control group (16.1 ±0.9)nmol·g-1 ·min-1,the activities of rat CYP3A in SLI 3,10 and 30 mg·kg-1 groups were 15.9 ±1.0,15.9±0.7 and (15.7 ±0.6)nmol·g-1 ·min-1 (P＜0.05).③ ATPase activity of SLI 0.0002,0.0006,0.0002,0.005,0.017,0.052,0.156 and 0.468 g·L-1 according to the clinical human blood-drug level was compared with that in control group,respectively (5.8,5.3,5.8,5.5,5.8,5.2,5.8,and 5.3 vs 5.8 μmol · g-1 · min-1),but there was no significant difference.CONCLUSION SLI may neither induce rat CYP1A2 or CYP3A in vivo,nor inhibit human CYP1A2,CYP2C9,CYP2C19,CYP2D6 and CYP3A4 in vitro.It is not a potential substrate or inhibitor for human P-gp in vitro.
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