刺五加注射液对大鼠肝微粒体四种CYP450亚型酶活性的影响

目的: 研究刺五加注射液在大鼠体外肝微粒体中对CYP2C9、CYP2D6、CYP2E1和CYP3A4活性的影响,为临床合理联合用药提供参考。方法: 在大鼠体外肝微粒体中分别加入四种亚型酶的探针药物甲苯磺丁脲(TB)、右美沙芬(DM)、氯唑沙宗(CLZ)、睾酮(TS)和低、中、高剂量的刺五加注射液,温孵后用HPLC法测定各空白对照组和不同剂量刺五加注射液给药组中各探针药物代谢产物的浓度并比较代谢率的差异,以评价刺五加注射液对各亚型酶活性的影响;中剂量组活性显著降低的亚型酶进一步考察抑制作用的强弱(即IC₅₀和K_i值)。结果: 与空白对照组比较,刺五加注射液低、中、高剂量给药组对CYP3A4活性的影响均有统计学意义(P<0.01),抑制率分别为 10.22%、19.00%、30.29%,其IC₅₀和K_i值分别为 3.96%和 2.74%(V/V);低、中、高剂量给药组对CYP2D6活性的影响均无统计学差异(P>0.05);低剂量给药组对CYP2C9、CYP2E1活性的影响无统计学差异(P>0.05),中、高剂量给药组对两个亚型的抑制作用有统计学差异(P<0.05),但中剂量给药组抑制率均小于8.50%,高剂量给药组抑制率均小于 12.00%。结论: 刺五加注射液对大鼠体外肝微粒体CYP3A4有抑制作用,且符合混合型抑制模型;对CYP2C9、CYP2E1抑制作用较弱;对CYP2D6活性无影响。

Effects of Ciwujia Injection on four cytochrome P450 isoforms in rat liver microsomes in vitro

AIM: To investigate the effects of Ciwujia Injection on CYP2C9, CYP2D6, CYP2E1 and CYP3A4 in rat liver microsomes in vitro, providing references for clinical rational use of drug combination therapy.METHODS: Probe drugs of tolbutamide(TB), dextromethorphan(DM), chlorzoxazone(CLZ) and testosterone(TS) as well as Ciwujia Injection of different concentrations were added to incubation systems of rat liver microsomes in vitro, the concentrations of metabolites of the probe drugs in blank control, low, medium and high dose of Ciwujia Injection groups were determined by HPLC after incubation, and the differences of metabolic rates were compared to evaluate the effects of Ciwujia Injection on the activities of the four subtypes.The strength of inhibition (IC₅₀ and K_i values) was further determined if the activity in the medium dose group significantly decreased.RESULTS: Compared with the blank control group, the effects on CYP3A4 activity were statistically significant in low, medium and high dose groups of Ciwujia Injection (P<0.01), and the inhibition rates were 10.22%, 19.00%, 30.29%, with the IC₅₀ and K_i values were 3.96% and 2.74%(V/V); the activities of CYP2D6 were not statistically significant in low, medium and high dose groups (P>0.05); the activities of CYP2C9, CYP2E1 were not statistically significant in low dose groups (P>0.05), meanwhile the inhibitory effects on the two subtype activities in medium and high dose groups were statistically significant (P<0.05), but the rates of inhibition were less than 8.50% in the medium dose groups and less than 12.00% in the high dose groups.CONCLUSION: Ciwujia Injection exhibited inhibition against the activity of CYP3A4 in rat liver microsomes in vitro, with mixed inhibition model, and weak inhibition on CYP2C9, CYP2E1, but no effect on CYP2D6.