不同急性肝损伤大鼠模型对5种CYP450酶的影响

Effect of Five Kinds of CYP450 Enzymes in Different Acute Liver Injury Rat Models

目的比较研究不同化学品致急性肝损伤大鼠模型体内CYP450酶活性的变化。方法以正常大鼠作为对照,分别采用四氯化碳(CCl_4)、D-氨基半乳糖(D-Gal N)/脂多糖(LPS)、α-萘异硫氰酸酯(ANIT)复制大鼠急性肝损伤模型,以Cocktail探针法,单剂量尾静脉注射由CYP450 5种同工酶的特异性探针底物非那西丁(CYP1A2)、甲苯磺丁脲(CYP2C9)、奥美拉唑(CYP2C19)、右美沙芬(CYP2D6)和咪达唑仑(CYP3A4)配置的Cocktail探针溶液后,从大鼠眼底静脉丛采集不同时间点的血样,采用UPLC-MS/MS检测5种探针药物的血药浓度,采用PK Solution 2^(TM)对数据进行处理,并计算主要药动学参数。与正常大鼠相比,不同模型大鼠的探针药物药动学行为的改变作为CYP450同工酶代谢活力评估依据。结果与正常大鼠相比:CCl_4模型大鼠的CYP1A2、CYP2C9、CYP2C19、CYP2D6与CYP3A4的活性均受到抑制,其中对CYP3A4的抑制作用比较轻微;D-Gal N/LPS模型大鼠的CYP2C9、CYP2D6与CYP3A4的活性均受到诱导,其中对CYP2D6与CYP3A4的诱导较轻微,CYP2C19的活性被抑制,但对CYP1A2的影响不明显;ANIT模型大鼠的CYP2C9、CYP2C19与CYP3A4的活性被诱导,对CYP3A4的诱导较轻微,CYP1A2的活性被强烈抑制,对CYP2D6作用不明显。结论不同化学品致急性肝损伤模型大鼠体内CYP450酶活性存在非常显著差异。

OBJECTIVE To compare the effect on CYP450 isoenzyme in rats with acute liver injury induced by different chemi- cals. METHODS Acute liver injury model of rats induced by tetrachloromethane （ CCl4 ）, D-aminogalactose （D-GaIN）/lipopolysac- charide （LPS）, α-naphthyl isothiocyanate （ANIT） respectively whereas the normal Wistar rats were used as controls. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates （phenacetin-CYP1A2, omeprazole-CYP2C9, tolbu- tamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4）, the blood samples were collected from the fundus venous plexus of rat at different time point, the blood drug concentration of the five probe substrates were determined by LC-MS/MS, and the pharma- cokinetic parameters were calculated by PK Solutions 2TM. Compared with normal rats, the changes of the probe drug pharmacokinetics in different rat models were used as the basis for the evaluation of the metabolic activity of CYP450 isoenzyme. RESULTS Compared with normal rats, the activities of CYP1A2, CYP2C9, CYP2C19, CYP2D6 of CC14 group rats were significantly inhibited,and the activities of CYP3A4 was slightly inhibited ; the activities of CYP2C9, CYP2D6 and CYP3A4 of D-GalN/LPS group rats were significantly induced, and the activity of CYP2D6 and CYP3A4 was slightly induced, and the activity of CYP1A2 was not significantly affected, but the activity of CYP2C19 was significantly inhibited; the activities of CYP2C9, CYP2C19 and CYP3A4 of ANIT group rats were significantly induced, the activity of CYP3A4 were slightly induced, and the activity of CYP2D6 was not significantly affected, but the activity of CYP1A2 was significantly inhibited. CONCLUSION There are significant differences in the activities of CYP450 isoenzyme in the rat model of acute liver injury induced by different chemicals.