尿毒清颗粒对CYP450与P-gp体外影响的初步研究

Effect of Niaoduqing granules on CYP450 and P-gp in vitro

关键词:  重组人细胞色素P450  P-糖蛋白三磷酸腺苷酶  半数抑制浓度  尿毒清颗粒

Key words:  CYP450  P-gp ATPase  IC₅₀  Niaoduqing granule

摘要:

[目的] 初步考察尿毒清颗粒(NDQ)对重组人细胞色素P450(CYP450)和P糖蛋白(P-gp)的体外影响.[方法] 1)使用 Promega P450-GloTM Screening System,通过荧光发光原理来检测NDQ组、抑制剂组对重组CYP450的IC₅₀(半数抑制浓度)值,比较NDQ组与抑制剂组IC₅₀来评价NDQ对重组人CYP1A2、CYP2D6、CYP3A4、CYP2C19,CYP2C9的抑制作用.2)使用BD ATPase Assay Kit,通过发光法检测NDQ组、空白对照组P-gp三磷酸腺苷(ATP)酶活性,比较NDQ组、空白对照组ATP酶活性来评价NDQ是否为重组人P-gp 的底物或抑制剂.[结果] 1)NDQ组与抑制剂组IC₅₀值(g/L)如下,CYP1A2: 17.47、3.37×10^-6;CYP3A4:33、4.181 3×10^-5; CYP2C9:10.39、1.056×10^-3; CYP2D6:13.98 、2.245 9×10^-6;CYP2C19:9.251、1.442×10^-3.2)NDQ组的P-gp ATP酶的活性为16.39 nmol/(mg·min),空白对照组的ATP酶活性为4.72 nmol/(mg·min).[结论] 1)NDQ组IC₅₀大于对应抑制剂组IC₅₀, NDQ对重组人CYP1A2、CYP2D6、CYP3A4、CYP2C19、CYP2C9基本没有抑制作用.2)NDQ组ATP酶活性高于空白组并差异有统计学意义,NDQ可能为重组人P-gp的抑制剂或底物.

Abstract:

[Objective] To investigate the effects of Niaoduqing (NDQ) on recombined human cytochrome P450 (CYP450) and P glycoprotein(P-gp) in vitro primarily. [Methods] 1)Using the Promega P450-GloTM Screening System and according to the fluorescence luminescence principle the IC₅₀ of NDQ group and inhibitor group to the IC₅₀ of recombined CYP450 was detected, comparing the IC₅₀ of NDQ group and inhibitor group the inhibitory effect of NDQ on recombined human CYP1A2, CYP2D6, CYP3A4, CYP2C19, CYP2C9 was evaluated. 2)Using the BD ATPase Assay Kit and using luminescent technique the activity of P-gp ATPase in the NDQ group and blank control group was detected and through the comparison of the ATPase activity between NDQ and blank control group it was evaluated that whether NDQ was the substrates or inhibitors of recombined human P-gp or not. [Results] 1)IC₅₀ value(g/L)in NDQ group and inhibitors group was as follows, CYP1A2：17.47, 3.37×10^-6;CYP3A4：33, 4.181×10^-5; CYP2C9：10.39, 1.056×10^-3;CYP2D6：13.98, 2.245 9×10^-6; CYP2C19：9.251, 1.442×10^-3. 2)The vbg htyu78 of P-gp ATPase activity in NDQ group was 16.39 nmol/(mg·min). ATPase activity in the blank control group was 4.72 nmol/(mg·min). [Conclusion] 1)IC₅₀ in NDQ group is greater than the corresponding inhibitor group. NDQ basically has no inhibitory effect on recombined human CYP1A2, CYP2D6, CYP3A4, CYP2C19, CYP2C9. 2)The activity of ATPase in NDQ group is higher than blank group significantly, and NDQ possibly is the inhibitor or the substrates of the recombined human P-gp.