甲基莲心碱对大鼠肝CYP450酶含量及CYP2D1,CYP3A1,CYP2E1mRNA的影响

Effect of Neferine on CYP450 Total Enzyme and CYP3A1,CYP2D1,CYP2E1 mRNA in Rats

目的:探讨甲基莲心碱对大鼠微粒体混悬液蛋白中细胞色素P450(CYP450)含量,及其对CYP450亚型CYP3A1,CYP2D1,CYP2E1的mRNA表达水平的影响。方法:Wistar大鼠48只,随机分成空白组、肝药酶诱导剂组、肝药酶抑制剂组,甲基莲心碱(Nef)低、中、高剂量组。对照组给予1%羧甲基纤维素钠(CMC-Na),诱导剂组地塞米松100mg·kg-1,抑制剂组酮康唑40mg·kg-1,Nef低、中、高剂量组分别为Nef10,20,50mg·kg-1灌胃给药,1日1次,连续6d。取大鼠的肝制备肝微粒体,测定肝微粒体中的蛋白浓度,用分光光度计对样品进行比色测定CYP450总酶的含量。并采用实时定量反转录--聚合酶链反应(Quantitive real-time RT-PCR)定量分析了大鼠CYP450亚型CYP3A1,CYP2D1,CYP2E1的mRNA表达水平。结果:甲基莲心碱在高剂量组的P450含量与空白组比较有显著性差异(P<0.01),提示该药在较大剂量时(20～50mg·kg-1)对CYP450有诱导作用。Nef在高剂量时,分别对CYP3A1和CYP2D1有诱导作用,可显著升高二者的活性(分别为空白组的1.92,2.99倍,P<0.05);中剂量时对CYP2D1具有诱导作用(为空白组的2.40倍,P<0.05);Nef在3个剂量下对CYP2E1均无诱导作用。结论:Nef在20,50mg·kg-1剂量可增加CYP450总酶活性和CYP2D6mRNA的表达,在50mg·kg-1剂量可增加CYP3A1mRNA的表达,提示Nef在较高剂量时可诱导肝药酶加速自身代谢。

Objective：to observe the impact of neferine on contents of CYP450 in rat liver and expression of CYP2D1,CYP3A1,CYP2E1 mRNA. Method：neferine was gived orally to Wistar rats for 6 days. Liver microsomes were separated with calcium carbonate precipitation and CYP450 enzyme content was measured with ultraviolet spectrophotometry. The expression of CYP2D1,CYP3A1,CYP2E1 mRNA was detected with Real-time quantitative RT-PCR. Result：①The result of Neferine on CYP450 total enzyme in rats indicated Neferine had inducing effect on drug-metabolizing enzyme at high dose; ②The expression levels of CYP3A1 and CYP2D1 were both increased significantly in the Neferine 50 mg·kg^ -1 group compared with the control group （P 〈0. 05）; ③The expression of CYP2D1 was also increased significantly in the Nef 20 mg·kg^ -1 group（P 〈0. 05）; ④There is no significant differences on expression of CYP2D1 in the all dosing Nef groups and the control group. These results indicated that Nef can raise the expression of CYP2D1 at 50 mg·kg ^-1 or 20 mg·kg^ -1,and raise the expression of CYP3A1 at 50 mg·kg^ -1. It inferred that Nef could accelerate its own metabolism. Conclusion：Neferine had effect on inducing CYP450 tatal enzyme at 20 or 50 mg ·kg^-1,and can raise the expression of CYP2D1. Neferine can also raise the expression of CYP3A1 at50 mg·kg^-1. It inferred that Neferine could accelerate its own metabolism at relative high doses in body.