热毒宁注射液对大鼠肝脏CYP450酶的影响

Effects of Reduning injection on activity of hepatic microsomal CYP450 isozymes in rats

研究热毒宁注射液对大鼠肝脏CYP450酶酶活性及其mRNA表达水平的影响。将SD大鼠腹腔注射7 d后,处死,制备大鼠的肝微粒体,与探针药物共孵育,利用LC/MS法测定各同工酶特异性底物在大鼠肝微粒的代谢产物的生成量,间接反映各同工酶的活性。应用荧光定量PCR技术考察热毒宁注射液对大鼠肝脏CYP450酶mRNA表达水平的影响。结果显示,酶活性方面,热毒宁注射液与空白组比较可抑制CYP2B1,2C12,2C13,2D2的酶活性,诱导CYP3A1的酶活性,对CYP1A2酶活性无明显影响。mRNA表达方面,热毒宁注射液对CYP2B1,3A1,1A2,2C11,2D1,2E1 mRNA的表达具有诱导作用。通过对上述结果分析,可以看出热毒宁注射液在酶活性和mRNA水平均对CYP3A1具有显著的诱导作用,与大环内酯类抗生素合用时可能会出现不良反应;对CYP2B1,2C12,2C13,2D2的酶活性均具有抑制作用,与其他药物联合用药时要充分考虑可能出现的药物相互作用或潜在风险。

To research the influence of Reduning injection on the activity and mRNA expression of cytochrome P450 （CYP450） system in rat liver microsomes. Rat liver microsomes were prepared after a seven-days continuous administration of Reduning injection. An HPLC-MS method was applied to determine the specific metabolites of CYP450 probe substrates in rat liver microsomal incubations. The activity of CYP450 isozymes were represented by the formation of metabolites. The Real-time quantitative polymerase chain reaction （Q-PCR） was applied to determine the mRNA expression levels of CYP450. Reduning injection significantly reduced the activity of CYP2B1,2C12,2C13 （P 〈 0. 01 ）, but did not affect CYP1A2 ; low dose and high dose of Reduning injection had an inhibition trend on the activity of CYP2D2, but did not statistically differ from control group ; low dose of Reduning injection significantly induced the ac- tivity of CYP3A1 （P 〈 0. 01 ）, high dose of Reduning injection had an induce trend on the activity of CYP3A1, but did not statistically differ from control. At the mRNA level, low and high dose of Reduning injection had an induce trend on the expression of CYP1A2, 2C11,2D1, 2El, 3A1, but did not statistically differ from control. Reduning injection significantly induced the activity of CYP2B1.Reduning injection significantly induced the activity of CYP3A1 in mRNA expression and enzyme activity levels, which may result ad- verse drug reaction after being combined with macrolides antibiotics. Reduning injection significantly reduced the activity of CYP2B1, 2C12, 2C13, 2D2 in enzyme activity levels, when combined with other drugs, it should be fully taken into account of the possible drug-drug interaction in order to avoid adverse side effects.