Cocktail法评价敌敌畏对大鼠CYP450亚型酶活性的影响
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摘要
目的用Cocktail探针药物法评估敌敌畏对大鼠CYP450酶代谢活性的影响。方法将大鼠随机分为敌敌畏组和对照组。敌敌畏组给予腹腔注射12.5 mg/kg敌敌畏,对照组给予等体积生理盐水。然后灌胃给予5种探针药物(安非他酮、美托洛尔、非那西丁、睾酮和甲苯磺丁脲),用超高效液相色谱串联质谱法测定血浆探针浓度;同时分离肝脏,用PCR法测定5种亚型酶mRNA的表达量。结果敌敌畏组CYP1A2、CYP2D1和CYP3A2的AUC明显高于对照组,差异有统计学意义(P<0.05),而对CYP2B1和CYP2C11的药代动力学无显著影响;CYP2D1和CYP3A2的mRNA表达水平均显著降低(P<0.05),CYP1A2的mRNA表达水平明显升高,差异有统计学意义(P<0.05),而CYP2C11和CYP2B1的mRNA表达水平与对照组比较差异无统计学意义(P>0.05)。结论结合PCR结果,腹腔注射敌敌畏可以抑制大鼠CYP2D1和CYP3A2的酶活性。
Objective In this work,a cocktail method was employed to evaluate the effects of dichlorvos on the metabolic activities of CYP450. Methods The rats were divided into dichlorvos group and control group. The dichlorvos group rats were given12. 5 mg/kg dichlorvos by intraperitoneal administration,and the control group were given normal saline. The probe drugs( bupropion,metroprolol,phenacetin,testosterone and tolbutamide) were given to rats by intragastric administration,and the ultra high performance liquid chromatography tandem mass spectrometry( UPLC-MS/MS) method was used to determined probe plasma concentrations. At the same time,livers were isolated and the mRNA expression levels of five CYP450 isoforms were determined by PCR. Results AUC of CYP1 A2,CYP2 D1 and CYP3 A2 in the dichlorvos group increased significantly( P < 0. 05),while pharmacokinetics difference for CYP2 B1 and CYP2 C11 had no significant changes. Expression levels of CYP2 D1 and CYP3 A2 mRNA declined significantly,with the difference statistically significant( P < 0. 05),while that of CYP1 A2 mRNA increased significantly,with the difference statistically significant( P < 0. 05). There was no statistical significance on the difference in expression levels of CYP2 C11 and CYP2 B1 mRNA by comparing dichlorvos group with control group( P > 0. 05). Conclusion Combined with PCR results,intraperitoneal administration of dichlorvos could inhibit the activities of CYP2 D1 and CYP3 A2 of rats.
Objective In this work,a cocktail method was employed to evaluate the effects of dichlorvos on the metabolic activities of CYP450. Methods The rats were divided into dichlorvos group and control group. The dichlorvos group rats were given12. 5 mg/kg dichlorvos by intraperitoneal administration,and the control group were given normal saline. The probe drugs( bupropion,metroprolol,phenacetin,testosterone and tolbutamide) were given to rats by intragastric administration,and the ultra high performance liquid chromatography tandem mass spectrometry( UPLC-MS/MS) method was used to determined probe plasma concentrations. At the same time,livers were isolated and the mRNA expression levels of five CYP450 isoforms were determined by PCR. Results AUC of CYP1 A2,CYP2 D1 and CYP3 A2 in the dichlorvos group increased significantly( P < 0. 05),while pharmacokinetics difference for CYP2 B1 and CYP2 C11 had no significant changes. Expression levels of CYP2 D1 and CYP3 A2 mRNA declined significantly,with the difference statistically significant( P < 0. 05),while that of CYP1 A2 mRNA increased significantly,with the difference statistically significant( P < 0. 05). There was no statistical significance on the difference in expression levels of CYP2 C11 and CYP2 B1 mRNA by comparing dichlorvos group with control group( P > 0. 05). Conclusion Combined with PCR results,intraperitoneal administration of dichlorvos could inhibit the activities of CYP2 D1 and CYP3 A2 of rats.
引文
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[19]Borkar RM,Bhandi MM,Dubey AP,et al.Plasma protein bindin pharmacokinetics,tissue distribution and CYP450 biotransformation studies of fidarestat by ultra high performance liquid chromatographyhigh resolution mass spectrometry[J].J Pharm Biomed Anal,2014,102C:386-399.
[20]Tan ML,Lim LE.The effects of Andrographis paniculata(Burm.f.)Nees extract and diterpenoids on the CYP450 isoforms'activities,a review of possible herb-drug interaction risks[J].Drug Chem Toxicol,2014,38(3):241-253.
[21]Wu Q,Zhang Q,Wen C,et al.The effect of MS-275 on CYP450isoforms activity in rats by cocktail method[J].Int J Clin Exp Pathol,2015,8(8):9360-9367.
[22]Videau O,Pitarque S,Troncale S,et al.Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat?[J].Xenobiotica,2012,42(4):349-354.
[23]Martignoni M,Groothuis GMM,de Kanter R.Species differences between mouse,rat,dog,monkey and human CYP-mediated drug metabolism,inhibition and induction[J].Expert Opin Drug Metab Toxicol,2006,2(6):875-894.
[24]ZacharˇováA,Siller M,SpicˇákováA,et al.Rosuvastatin suppresses the liver microsomal CYP2C11 and CYP2C6 expression in male Wistar rats[J].Xenobiotica,2012,42(8):731-736.
[2]Celik MM,Alp A,Dokuyucu R,et al.Protective Effects of Intralipid and Caffeic Acid Phenethyl Ester on Nephrotoxicity Caused by Dichlorvos in Rats[J].J Anal Methods Chem,2015:491406.
[3]Nelson DR,Kamataki T,Waxman DJ,et al.The P450 superfamily:update on new sequences,gene mapping,accession numbers,early trivial names of enzymes,and nomenclature[J].DNA Cell Biol,1993,12(1):1-51.
[4]Tan ML,Lim LE.The effects of Andrographis paniculata(Burm.f.)Nees extract and diterpenoids on the CYP450 isoforms'activities,a review of possible herb-drug interaction risks[J].Drug Chem Toxicol,2015,38(3):241-253.
[5]Ragia G,Giannakopoulou E,Karaglani M,et al.Frequency of CYP450 enzyme gene polymorphisms in the Greek population:review of the literature,original findings and clinical significance[J].Drug Metabol Drug Interact,2014,29(4):235-248.
[6]Zhang L,Reynolds KS,Zhao P,et al.Drug interactions evaluation:an integrated part of risk assessment of therapeutics[J].Toxicol Appl Pharmacol,2010,243(2):134-145.
[7]Breimer DD,Schellens JH.A'cocktail'strategy to assess in vivo oxidative drug metabolism in humans[J].Trends Pharmacol Sci,1990,11(6):223-225.
[8]Wang X,Chen X,Chen M,et al.Assessment of effects of chronic hydrogen sulfide poisoning on cytochrome P450 isoforms activity of rats by cocktail approach[J].Biol Pharm Bull,2013,36(10):1627-1633.
[9]Wang X,Chen M,Chen X,et al.The effects of acute hydrogen sulfide poisoning on cytochrome P450 isoforms activity in rats[J].Biomed Res Int,2014:209393.
[10]Gunay N,Kose B,Demiryurek S,et al.Protective effects of Y-27632 on acute dichlorvos poisoning in rats[J].Am J Emerg Med,2010,28(3):268-274.
[11]Wang NN,Yuan L,Dai H,et al.Effect of PON1 on dichlorvos toxicokinetics[J].Emerg Med J,2011,28(4):313-315.
[12]Huang Y,Zheng SL,Zhu HY,et al.Effects of aescin on cytochrome P450 enzymes in rats[J].J Ethnopharmacol,2014,151(1):583-590.
[13]Xu RA,Xu ZS,Ge RS.Effects of hydroxysafflor yellow A on the activity and mRNA expression of four CYP isozymes in rats[J].J Ethnopharmacol,2014,151(3):1141-1146.
[14]Lin K,Zhang Q,Liu Z,et al.Effects of suberoylanilide hydroxamic acid on rat cytochrome P450 enzyme activities[J].Int J Clin Exp Pathol,2015,8(5):5584-5590.
[15]Ma J,Wang S,Zhang M,et al.Simultaneous determination of bupropion,metroprolol,midazolam,phenacetin,omeprazole and tolbutamide in rat plasma by UPLC-MS/MS and its application to cytochrome P450 activity study in rats[J].Biomed Chromatogr,2015,29(8):1203-1212.
[16]Su T,Mao C,Yin F,et al.Effects of unprocessed versus vinegarprocessed Schisandra chinensis on the activity and mRNA expression of CYP1A2,CYP2E1 and CYP3A4 enzymes in rats[J].J Ethnopharmacol,2013,146(3):734-743.
[17]Lin GY,Ma JS,Xu RA,et al.Effects of Ougan juice on P450 activities using a cocktail method[J].Pharmazie,2012,67(3):242-246.
[18]Qin CZ,Ren X,Tan ZR,et al.A high-throughput inhibition screening of major human cytochrome P450 enzymes using an in vitro cocktail and liquid chromatography-tandem mass spectrometry[J].Biomed Chromatogr,2014,28(2):197-203.
[19]Borkar RM,Bhandi MM,Dubey AP,et al.Plasma protein bindin pharmacokinetics,tissue distribution and CYP450 biotransformation studies of fidarestat by ultra high performance liquid chromatographyhigh resolution mass spectrometry[J].J Pharm Biomed Anal,2014,102C:386-399.
[20]Tan ML,Lim LE.The effects of Andrographis paniculata(Burm.f.)Nees extract and diterpenoids on the CYP450 isoforms'activities,a review of possible herb-drug interaction risks[J].Drug Chem Toxicol,2014,38(3):241-253.
[21]Wu Q,Zhang Q,Wen C,et al.The effect of MS-275 on CYP450isoforms activity in rats by cocktail method[J].Int J Clin Exp Pathol,2015,8(8):9360-9367.
[22]Videau O,Pitarque S,Troncale S,et al.Can a cocktail designed for phenotyping pharmacokinetics and metabolism enzymes in human be used efficiently in rat?[J].Xenobiotica,2012,42(4):349-354.
[23]Martignoni M,Groothuis GMM,de Kanter R.Species differences between mouse,rat,dog,monkey and human CYP-mediated drug metabolism,inhibition and induction[J].Expert Opin Drug Metab Toxicol,2006,2(6):875-894.
[24]ZacharˇováA,Siller M,SpicˇákováA,et al.Rosuvastatin suppresses the liver microsomal CYP2C11 and CYP2C6 expression in male Wistar rats[J].Xenobiotica,2012,42(8):731-736.