基于CYP3A4酶探讨灯盏花素对洛伐他汀的药动学的影响及机制

Effects of Breviscapine on Pharmacokinetics of Lovastatin Based on CYP3A4 Enzyme and It’s Mechanism

关键词:  灯盏花素  洛伐他汀  药动学  CYP3A4  RT-PCR

Key words:  breviscapine  lovastatin  pharmacokinetic  CYP3A4  RT-PCR

摘要:

目的 探究灯盏花素与洛伐他汀联用对大鼠体内药动学的影响，从代谢酶的角度揭示灯盏花素对洛伐他汀药动学产生影响的机制。方法 采用探针药物法及HPLC测定咪达唑仑在肝微粒体孵育体系中的浓度，评价灯盏花素与洛伐他汀联用对CYP3A4酶活性的影响。通过RT-PCR反应来检测CYP3A4酶mRNA基因表达，采用Western blotting，从蛋白翻译水平上分析灯盏花素与洛伐他汀联用对大鼠肝脏CYP3A4蛋白表达的影响。结果 洛伐他汀与灯盏花素联合用药后，洛伐他汀在大鼠体内的血药浓度显著升高，从0.39 mg·L^-1升到1.08 mg·L^-1，清除率从3.36 L·h^-1·kg^-1降低到1.86 L·h^-1·kg^-1，药物半衰期从5.0 h延长到6.3 h，联合给药后洛伐他汀的AUC从2.43 mg·h·L^-1上升到4.22 mg·h·L^-1。与空白组比较，洛伐他汀组的CYP3A4酶活性没有明显变化；灯盏花素组及灯盏花素+洛伐他汀组与空白组比较发现均明显抑制CYP3A4酶活性；灯盏花素与灯盏花素+洛伐他汀组CYP3A4酶mRNA表达量均较空白组显著降低；CYP3A4酶蛋白含量结果表明，洛伐他汀组、灯盏花素+洛伐他汀组与空白组比较CYP3A4酶蛋白含量均没有明显变化。结论 洛伐他汀与灯盏花素联用，灯盏花素通过抑制其基因转录水平抑制CYP3A4的活性，使大鼠体内药动学过程发生变化，使洛伐他汀药物的代谢减慢。

Abstract:

OBJECTIVE To investigate the effect of the breviscapine combined with lovastatin on the pharmacokinetics of rats in vivo and to reveal the mechanism of the effect of breviscapine on the pharmacokinetics of lovastatin from the perspective of metabolic enzyme. METHODS The concentration of midazolam in liver microsome incubation system was determined by probe drug and HPLC. The effect of breviscapine combined with lovastatin on the activity of CYP3A4 was evaluated. The mRNA expression of CYP3A4 was detected by RT-PCR. And the effect of breviscapine combined with lovastatin on the expression of CYP3A4 protein in rat liver was analyzed at the level of protein translation by Western blotting. RESULTS After the treatment of combination of lovastatin and breviscapine, the plasma concentration of lovastatin in rats increased significantly from 0.39 mg·L^-1 to 1.08 mg·L^-1, clearance decreased from 3.36 L·h^-1·kg^-1 to 1.86 L·h^-1·kg^-1, the drug half-life was extended from 5.0 h to 6.3 h, the AUC of lovastatin after combined administration was 2.43 mg·h·L^-1rose to 4.22 mg·h·L^-1. Compared with the blank group, activity of CYP3A4 did not change significantly in the lovastatin group. Breviscapine group and breviscapine combined with lovastatin group significantly inhibited CYP3A4 activity compared with the blank group; Breviscapine group and breviscapine combined with lovastatin group decreased CYP3A4 enzyme expression significantly compared with the blank group. CYP3A4 enzyme protein content showed that, compared with the blank group, there was no significant change in CYP3A4 enzyme protein content in lovastatin group and breviscapine combined with lovastatin group. CONCLUSION After the combination of lovastatin and breviscapine, breviscapine inhibite CYP3A4 activity by inhibiting the gene transcription level of the enzyme. The combination change the pharmacokinetic process in rats and slow down the metabolism of lovastatin.