健脾化浊方对阿托伐他汀体外代谢的影响

目的：在体外基于细胞色素（CYP450）酶系研究健脾化浊方对阿托伐他汀代谢的影响. 方法：首先将健脾化浊方与人肝微粒体预孵育15 min后,加入CYP 3A4的底物睾酮及还原型辅酶烟酰胺腺嘌呤二核苷酸,于37℃共同孵育30 min,之后加入0.5 ml预冷的甲醇终止反应,取等量上清于高效液相质谱分析仪上检测6β-羟基睾酮的生成,确立体外肝微粒体中健脾泄浊方对CYP3A4酶的作用.再将稀释好的健脾化浊方及人肝微粒体加入96深孔板中,然后加入阿托伐他汀,对照组中则加入阴性对照物,取上清用于液相色谱/质谱/质谱分析. 结果：健脾化浊方对肝药酶CYP3A4有较强的抑制作用（IC50=10.5）,并且能延缓阿托伐他汀在肝细胞微粒体内的代谢,使其药物浓度显著提高（P＜0.01）. 结论：体外健脾化浊方通过对CYP3A4酶的抑制作用而影响了阿托伐他汀的代谢,中药和西药联用存在着药物相互作用.

Effect of tonifying spleen and eliminating turbidity decoction on atorvastatin metabolism in vitro

Objective：To evaluate the effect of tonifying spleen and eliminating turbidity decoction （TSET） on atorvastatin metabolism based on CYP450 enzyme in vitro. Methods： Firstly, different concentrations of TSET were co-incubated with liver microsomes for 15 min at 37℃, then testosterone, the substrate of CYP3A4, and NADPH was added to incubate for another 30 min at 37℃. The reaction was terminated with pre-cooled 0.5 ml methanol, and then 6β-hydroxyl testosterone in the supernatant was measured by high performance liquid chromatography-mass spectrometry technology （HPLC-MS）. Secondly, atorvastatin was incubated with or without TSET and liver microsomes, and concentration of atorvastatin in supernatant was determined by HPLC-MS at a series of time points. Results：With the increase of TSET concentration, the activity of liver metabolizing enzyme CYP3A4 in liver microsomes was significantly inhibited （IC50 = 10.5）. TSET significantly increased the concentration of atorvastatin when incubated together with liver microsomes （P〈0.01）. Conclusion：TSET reduces the metabolism of atorvastatin in liver microsomes possibly through inhibiting the activity of CYP3A4. TSET and atorvastatin may have remarkable synergy effect.