基于经皮给药研究制川乌与白芍配伍对大鼠皮肤及肝微粒体中CYP450活性的影响

目的：研究经皮给予制川乌-白芍药对后,配伍药对大鼠皮肤、肝脏药物代谢酶的影响,以探讨制川乌-白芍药对的配伍机制。方法：大鼠连续经皮给药制川乌、白芍、制川乌与白芍配伍凝胶剂14 d后,采用探针底物与皮肤、肝微粒体体外孵育,通过高效液相色谱法（HPLC）检测皮肤微粒体中1种探针底物及肝微粒体中4种探针底物的浓度,计算代谢速率,考察各给药组大鼠皮肤、肝微粒体CYP450酶活性的变化。结果：与空白组比较,白芍、制川乌对大鼠皮肤微粒体中的CYP3A4酶具有明显的抑制作用（P<0.05）;与白芍单用组比较,制川乌与白芍配伍给药使大鼠皮肤微粒体中的CYP3A4酶活性显著下降（P<0.05）;与制川乌组比较,制川乌与白芍配伍给药使大鼠皮肤中微粒体的CYP3A4酶的活性显著增加（P<0.05）。与空白组比较,白芍、制川乌对大鼠肝微粒体中的CYP3A4、CYP1A2、CYP2E1、CYP2D6酶都具有明显的抑制作用（P<0.05）;与制川乌组、白芍组比较,制川乌与白芍配伍使大鼠肝微粒体中CYP3A4、CYP1A2、CYP2E1、CYP2D6酶活性均明显增加（P<0.05）。结论：制川乌、白芍均为CYP3A4、CYP1A2、CYP2E1、CYP2D6药酶的抑制剂,两药配伍可引起大鼠皮肤、肝微粒体中药酶活性的改变而致活性成分的代谢速度改变,这可能是制川乌与白芍配伍发挥"增效减毒"的作用机制之一。

Effects of compatibility of Aconiti radix cocta and Paeoniae radix alba on microsome cytochrome P450 activity of skin and liver in rats based on percutaneous administration

OBJECTIVE To observe the changes of skin metabolism enzymes and hepatic microsomal enzymes of rat treated with Aconiti radix cocta combined with Paeoniae radix alba gel to explore the compatibility mechanism.METHODS Rats were treated with continuous transdermal administration of Aconiti radix cocta gel, Paeoniae radix alba gel, Aconiti radix cocta combined with Paeoniae radix alba gel for 14 days. Probe substrate, liver microsomes or skin microsomes in vitro incubated method were adopted, the concentrations of four probe substrates in the liver microsomes and one probe substrate in the skin microsomes were determined by HPLC, metabolic rates were calculated, to study the effects of each administration group on the microsome cytochrome P450 activities of rat skin and liver.RESULTS Compared with the blank group, Aconiti radix cocta and Paeoniae radix alba obviously inhibited CYP3A4 enzyme in rat skin (P<0.05). Compared with the Paeoniae radix alba group, the activity of CYP3A4 enzyme was significantly decreased after preparation (P<0.05). While the activity of CYP3A4 enzyme was significantly increased compared with the Aconiti radix cocta group (P<0.05). Compared with the blank group, Aconiti radix cocta and Paeoniae radix alba administered alone obviously inhibited CYP3A4, CYP1A2, CYP2E1 and CYP2D6 enzymes in rat liver (P<0.05), meanwhile the activities of CYP3A4, CYP1A2, CYP2E1 and CYP2D6 enzyme were significantly increased after preparation (P<0.05).CONCLUSION Both Aconiti radix cocta and Paeoniae radix alba are the inhibitors of CYP3A4, CYP1A2, CYP2E1 and CYP2D6. Combination of the two drugs can change the enzyme activities of the skin and liver, and cause the changes of the metabolic rates of the active components of Raconiti radix cocta and Paeoniae radix alba. It suggests that the inhibitory and inducing effects of prepared Aconiti radix cocta and Paeoniae radix alba on metabolic enzymes may be one reason for their attenuation and synergistic effects.