桑叶提取物对大鼠体内细胞色素P450酶活性的影响
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摘要
目的采用"Cocktail"混合探针药物法,探究桑叶水提物(aqueous extract of mulberry leaves,AML)、桑叶醇提物(ethanol extract of mulberry leaves,EML)对大鼠体内细胞色素P450(cytochrome P450,CYP450)酶活性的影响,从代谢酶的角度预测它可能引起的药物相互作用,为临床合理用药提供参考。方法选用非那西丁、安非他酮、双氯酚酸钠作为大鼠体内3种CYP450酶(CYP1A2、CYP2B6、CYP2C9)的探针药物。雄性SD大鼠连续灌胃桑叶提取物14 d,第14天灌胃后30 min尾静脉注射混合探针药物,根据不同时间点采血。采用超高效液相色谱-串联质谱测定各组血药浓度,用DAS 2.0和SPSS 21.0软件拟合药物代谢动力学参数并进行统计学分析。结果连续灌胃桑叶提取物14 d后,与9.0 g/L氯化钠注射液相比,AML组中非那西丁、双氯酚酸钠代谢减慢,安非他酮则不明显;与羧甲基纤维素钠组相比,EML组中安非他酮、双氯酚酸钠代谢减慢,非那西丁则不明显。结论 AML对大鼠体内CYP1A2、CYP2C9可能存在一定的抑制作用,对CYP2B6可能无显著性影响;EML对大鼠体内CYP2B6、CYP2C9可能存在一定的抑制作用,对CYP1A2可能无显著性影响。
Objective To investigate the effects of aqueous extract of mulberry leaves(AML) and ethanol extract of mulberry leaves(EML) on the activity of cytochrome P450 enzymes in rats based on the cocktail probe drug method, as well as possible drug interactions caused by AML and EML from the aspect of metabolic enzymes, and to provide a reference for rational drug use in clinical practice. Methods Phenacetin, bupropion, and diclofenac sodium were selected as the probe drugs for three cytochrome P450 enzymes in rats(CYP1 A2, CYP2 B6, and CYP2 C9). Male Sprague-Dawley rats were given mulberry leaf extract by gavage for 14 consecutive days, and tail vein injection of probe drugs was performed at 30 minutes after the last administration on day 14. Blood samples were collected at different time points. Liquid chromatography-tandem mass spectrometry was used to measure the plasma concentrations of drugs, and DAS 2.0 and SPSS 21.0 were used for the fitting of pharmacokinetic parameters and were used to perform a statistical analysis. Results Compared with the 9.0 g/L sodium chloride injection group, the AML group had significant reductions in the metabolic rates of phenacetin and diclofenac sodium after 14 consecutive days of intragastric administration of AML, while there was no significant change in the metabolic rate of bupropion. Compared with the sodium carboxymethyl cellulose group, the EML group had significant reductions in the metabolic rates of bupropion and diclofenac sodium, while there was no significant change in the metabolic rate of phenacetin. Conclusion AML may have a certain inhibitory effect on the activities of CYP1 A2 and CYP2 C9 in rats, and may have no marked influence on the activity of CYP2 B6. EML may have a certain inhibitory effect on the activities of CYP2 B6 and CYP2 C9 in rats, and may have no marked influence on the activity of CYP1 A2.
Objective To investigate the effects of aqueous extract of mulberry leaves(AML) and ethanol extract of mulberry leaves(EML) on the activity of cytochrome P450 enzymes in rats based on the cocktail probe drug method, as well as possible drug interactions caused by AML and EML from the aspect of metabolic enzymes, and to provide a reference for rational drug use in clinical practice. Methods Phenacetin, bupropion, and diclofenac sodium were selected as the probe drugs for three cytochrome P450 enzymes in rats(CYP1 A2, CYP2 B6, and CYP2 C9). Male Sprague-Dawley rats were given mulberry leaf extract by gavage for 14 consecutive days, and tail vein injection of probe drugs was performed at 30 minutes after the last administration on day 14. Blood samples were collected at different time points. Liquid chromatography-tandem mass spectrometry was used to measure the plasma concentrations of drugs, and DAS 2.0 and SPSS 21.0 were used for the fitting of pharmacokinetic parameters and were used to perform a statistical analysis. Results Compared with the 9.0 g/L sodium chloride injection group, the AML group had significant reductions in the metabolic rates of phenacetin and diclofenac sodium after 14 consecutive days of intragastric administration of AML, while there was no significant change in the metabolic rate of bupropion. Compared with the sodium carboxymethyl cellulose group, the EML group had significant reductions in the metabolic rates of bupropion and diclofenac sodium, while there was no significant change in the metabolic rate of phenacetin. Conclusion AML may have a certain inhibitory effect on the activities of CYP1 A2 and CYP2 C9 in rats, and may have no marked influence on the activity of CYP2 B6. EML may have a certain inhibitory effect on the activities of CYP2 B6 and CYP2 C9 in rats, and may have no marked influence on the activity of CYP1 A2.
引文
[1] 高学敏.中药学[M].北京:人民卫生出版社,2000.
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[3] 李向荣,陈菁菁,刘晓光.桑叶总黄酮对高脂血症动物的降血脂效应[J].中国药学杂志,2009,44(21):1630-1633.
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[14] 蔡小军,黄凯,宋惠珠,等.斑蝥酸钠维生素B6和复方苦参注射液对人肝微粒体细胞色素P450酶活性的抑制作用[J].中国临床药理学杂志,2017,33(21):2150-2153.
[15] 崔明宇,王美薇,杨柳,等.蓬子菜香叶木苷对大鼠细胞色素P450酶活性影响[J].哈尔滨商业大学学报(自然科学版),2016,32(4):406-408.
[16] 洪小凤,林晶,杨强丽.“Cocktail”探针药物法评价苦碟子注射液对大鼠CYP450酶活性的影响[J].海峡药学,2017,29(3):25-29.
[17] 杨鑫宝,刘建勋.近10年中药与药物代谢酶相互作用的研究进展[J].中国中药杂志,2012,37(7):871-877.
[18] 秦琰杰,汪难喜,张新林,等.药物代谢酶和转运体介导的辣椒素-药物相互作用研究进展[J].中国药学杂志,2018,53(1):1-5.
[19] 国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:298.
[20] 王妍妍.茯苓对大鼠细胞色素P450酶活性及mRNA表达的影响[D].合肥:安徽中医药大学,2017.
[21] 洪世忠,施哲媛,何帆,等.左归饮水提物对大鼠Cyp2b1、Cyp2c11、Cyp2c7亚酶活性的影响[J].安徽中医药大学学报,2018,37(4):59-64.
[22] 葛韬,彭慧,张变,等.聚氧乙烯脱水山梨醇单油酸酯对大鼠体内细胞色素P450酶活性的影响[J].中南药学,2017,15(12):1699-1703.
[23] JIANG B,MENG L,ZHANG F,et al.Enzyme-inducing effects of berberine on cytochrome P450 1A2 in vitro and in vivo[J].Life Sciences,2017,189:1-7.
[24] KAPOOR N,PANT A B,DHAWAN A,et al.Cytochrome P450 1A isoenzymes in brain cells:expression and inducibility in cultured rat brain neuronal and glial cells[J].Life Sciences,2006,79(25):2387-2394.
[25] HE X,FENG X.Role of metabolic enzymes P450 (CYP) on activating procarcinogen and their polymorphisms on the risk of cancers[J].Current Drug Metabolism,2015,16(10):850-863.
[26] MARTIGNONI M,GROOTHUIS G M,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.
[2] 唐明敏.桑叶降糖有效物质基础及其作用机理研究[D].北京:北京中医药大学,2016.
[3] 李向荣,陈菁菁,刘晓光.桑叶总黄酮对高脂血症动物的降血脂效应[J].中国药学杂志,2009,44(21):1630-1633.
[4] MICHIELAN L,TERFLOTH L,GASTEIGER J,et al.Comparison of multilabel and single-label classification applied to the prediction of the isoform specificity of cytochrome p450 substrates[J].J Chem Inf Model,2009,49(11):2588-2605.
[5] WILLIAMS C A,HARBORNE J B,NEWMAN M,et al.Chrysin and other leaf exudate flavonoids in the genus Pelargonium[J].Phytochemistry,1997,46(8):1349-1357.
[6] 胡道德,顾磊,姚慧娟,等.Cocktail探针药物评价淫羊藿提取物对大鼠CYP450酶的影响[J].中国新药与临床杂志,2009,28(5):337-341.
[7] 樊慧蓉,和凡,刘昌孝.Cocktail探针药物法用于评价细胞色素P450同工酶影响的研究进展[J].中国药学杂志,2006,41(14):1045-1048.
[8] 陆兔林,苏联麟,季德,等.CYP450酶与中药代谢相互作用及酶活性测定的研究进展[J].中国中药杂志,2015,40(18):3524-3529.
[9] 陈伟,何瑞曦,彭慧,等.新藤黄酸对大鼠肝微粒体细胞色素P450亚型酶活性的影响[J].安徽中医药大学学报,2016,35(1):78-83.
[10] 董宇,王阶,杨庆,等.CYP450酶与中药代谢相互作用关系研究概况[J].中国中医药信息杂志,2011,18(1):100-103.
[11] 宋菲,杨健,王春,等.Cocktail 探针药物法评价羊耳菊对大鼠CYP450酶活性的影响[J].中国实验方剂学杂志,2018,24(19):29-36.
[12] 刘育含,才谦.无梗五加果实提取物对大鼠CYP450活性的影响[J].现代药物与临床,2017,32(8):1410-1415.
[13] 刘亭,刘香香,陆定艳,等.灯盏细辛和赤芍配伍组方对小鼠部分肝细胞色素P450亚型活性的影响及其机制分析[J].中国实验方剂学杂志,2018,24(13):124-130.
[14] 蔡小军,黄凯,宋惠珠,等.斑蝥酸钠维生素B6和复方苦参注射液对人肝微粒体细胞色素P450酶活性的抑制作用[J].中国临床药理学杂志,2017,33(21):2150-2153.
[15] 崔明宇,王美薇,杨柳,等.蓬子菜香叶木苷对大鼠细胞色素P450酶活性影响[J].哈尔滨商业大学学报(自然科学版),2016,32(4):406-408.
[16] 洪小凤,林晶,杨强丽.“Cocktail”探针药物法评价苦碟子注射液对大鼠CYP450酶活性的影响[J].海峡药学,2017,29(3):25-29.
[17] 杨鑫宝,刘建勋.近10年中药与药物代谢酶相互作用的研究进展[J].中国中药杂志,2012,37(7):871-877.
[18] 秦琰杰,汪难喜,张新林,等.药物代谢酶和转运体介导的辣椒素-药物相互作用研究进展[J].中国药学杂志,2018,53(1):1-5.
[19] 国家药典委员会.中华人民共和国药典:一部[M].北京:中国医药科技出版社,2015:298.
[20] 王妍妍.茯苓对大鼠细胞色素P450酶活性及mRNA表达的影响[D].合肥:安徽中医药大学,2017.
[21] 洪世忠,施哲媛,何帆,等.左归饮水提物对大鼠Cyp2b1、Cyp2c11、Cyp2c7亚酶活性的影响[J].安徽中医药大学学报,2018,37(4):59-64.
[22] 葛韬,彭慧,张变,等.聚氧乙烯脱水山梨醇单油酸酯对大鼠体内细胞色素P450酶活性的影响[J].中南药学,2017,15(12):1699-1703.
[23] JIANG B,MENG L,ZHANG F,et al.Enzyme-inducing effects of berberine on cytochrome P450 1A2 in vitro and in vivo[J].Life Sciences,2017,189:1-7.
[24] KAPOOR N,PANT A B,DHAWAN A,et al.Cytochrome P450 1A isoenzymes in brain cells:expression and inducibility in cultured rat brain neuronal and glial cells[J].Life Sciences,2006,79(25):2387-2394.
[25] HE X,FENG X.Role of metabolic enzymes P450 (CYP) on activating procarcinogen and their polymorphisms on the risk of cancers[J].Current Drug Metabolism,2015,16(10):850-863.
[26] MARTIGNONI M,GROOTHUIS G M,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.