土忍翘薇合剂对大鼠CYP450酶活力和mRNA的调控作用
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摘要
目的:研究土忍翘薇合剂对大鼠细胞色素P450(CYP450)酶活力和m RNA表达的调节作用。方法:20只SD大鼠随机分为实验组和对照组,分别连续灌胃土忍翘薇合剂或纯净水14 d后,同时给予探针药非那西丁、安非他酮、氯唑沙宗和咪达唑仑,并用液相色谱-质谱联用技术(液质联用)测定给药后不同时间点血药浓度,计算药动学参数进而推测CYP1A2、CYP2B1、CYP2E1和CYP3A1酶活力变化;实时荧光定量PCR(RTPCR)用于测定CYP450的m RNA表达水平。结果:与对照组比较,实验组大鼠非西丁的清除率提高了42%,曲线下面积减少了31%,达峰浓度减少了33%(P<0.05),说明土忍翘薇合剂对CYP1A2活力具有显著诱导作用;实验组咪达唑仑的清除率提高了66%,曲线下面积和达峰浓度分别为对照组的57%和52%,表观分布容积增加了1.36倍(P<0.05),说明土忍翘薇合剂对CYP3A1活力具有显著诱导作用;安非他酮和氯唑沙宗药时曲线图和药动学参数均提示土忍翘薇合剂对CYP2B1和CYP2E1活力无明显影响。RT-PCR检测发现,实验组大鼠CYP1A2和CYP3A1 m RNA表达分别上调为对照组的2.26倍和1.94倍(P<0.05),而CYP2B1与CYP2E1 m RNA表达2组间比较差异无统计学意义(P>0.05)。结论:土忍翘薇合剂会上调CYP1A2和CYP3A1酶活力及其m RNA表达水平。
Objective: To investigate the modulation of Tu Ren Qiao Wei decoction on activities and m RNA expressions of cytochrome P450(CYP450) enzymes in rats. Methods: Twenty SD rats were randomly divided into treatment group and control group. After treatment with Tu Ren Qiao Wei decoction or water for 14 days, treatment group rats and control group rats were given the mixture of four probe substrates including phenacetin, bupropion, chlorzoxazone and midazolam. The plasma concentration of probes at a series of time-points were determined by liquid chromatography tandom mass spectrometry. The activities of CYP450 s were evaluated according to the pharmacokinetic parameters of corresponding substrates. Real-time PCR was applied to assess the m RNA expression levels of CYP450 s. Results: The pharmacokinetic parameters of phenacetin and midazolam from treatment group showed significant differences compared with control group, which indicated that Tu RenQiao Wei decoction induced CYP1 A2 and CYP3 A1 activity. And no significant difference was found in pharmacokinetic parameters of bupropion or chlorzoxazone from treatment group and control group. In addition, treatment with Tu Ren Qiao Wei decoction significantly up-regulated the m RNA expression of CYP1 A2 and CYP3 A1 whereas it had no impact on CYP2 B1 and CYP2 E1. Conclusion: Tu Ren Qiao Wei decoction can up-regulate the activities and m RNA expressions of CYP1 A2 and CYP3 A1.
Objective: To investigate the modulation of Tu Ren Qiao Wei decoction on activities and m RNA expressions of cytochrome P450(CYP450) enzymes in rats. Methods: Twenty SD rats were randomly divided into treatment group and control group. After treatment with Tu Ren Qiao Wei decoction or water for 14 days, treatment group rats and control group rats were given the mixture of four probe substrates including phenacetin, bupropion, chlorzoxazone and midazolam. The plasma concentration of probes at a series of time-points were determined by liquid chromatography tandom mass spectrometry. The activities of CYP450 s were evaluated according to the pharmacokinetic parameters of corresponding substrates. Real-time PCR was applied to assess the m RNA expression levels of CYP450 s. Results: The pharmacokinetic parameters of phenacetin and midazolam from treatment group showed significant differences compared with control group, which indicated that Tu RenQiao Wei decoction induced CYP1 A2 and CYP3 A1 activity. And no significant difference was found in pharmacokinetic parameters of bupropion or chlorzoxazone from treatment group and control group. In addition, treatment with Tu Ren Qiao Wei decoction significantly up-regulated the m RNA expression of CYP1 A2 and CYP3 A1 whereas it had no impact on CYP2 B1 and CYP2 E1. Conclusion: Tu Ren Qiao Wei decoction can up-regulate the activities and m RNA expressions of CYP1 A2 and CYP3 A1.
引文
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[2]刘高峰,郭兴蕾.中药对细胞色素P450调控作用的研究进展[J].中草药,2008,39(1):139-143.
[3]陈熠.中医临床家陈苏生[M].1版.北京:中国中医药出版社,2001:141-142.
[4]陈熠,陈明华,陈建平.陈苏生医集纂要[M].1版.上海:上海科学技术文献出版社,1994:150-152.
[5]王建平,张海燕,傅旭春.土茯苓的化学成分和药理作用研究进展[J].海峡药学,2013,25(1):42-44.
[6]贾献慧,李静,张永清.忍冬藤化学成分研究进展[J].山东中医杂志,2015,34(8):641-643.
[7]宋小俊.连翘不同部位化学成分研究进展[J].西北药学杂志,2014,29(2):220-222.
[8]袁鹰,张卫东,柳润辉,等.白薇的化学成分和药理研究进展[J].药学实践杂志,2007,25(1):6-9.
[9]周子晔,金辉,王陈翔,等.绿茶水提物对大鼠CYP450酶活力和m RNA表达的调控作用[J].温州医科大学学报,2015,45(7):502-506,511.
[10]LEWIS D F V.P450 substrate specificity and metabolism.in:Lewis.Cytochrome P450:structure,function and mechanism[M].Third Edition.Bristol:Taylor&Francis,1996:102-152.
[11]ZHANG S,SONG N,LI Q,et al.Liquid chromatography/tandem mass spectrometry method for simultaneous evaluation of activities of five cytochrome P450s using a five-drug cocktail and application to cytochrome P450 phenotyping studies in rats[J].J Chromatogr B Analyt Technol Biomed Life Sci,2008,871(1):78-89.
[12]JURICA J,KYR M,MCCASKEY HADASOVA E,et al.Evaluation of the activity of P450 enzymes in rats:use of the single marker or combined drug administration[J].Neuro Endocrinol Lett,2009,30(1):92-95.
[13]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.
[14]肖鹏,周宏灏.细胞色素氧化酶CYP1A2的研究进展[J].中南大学学报(医学版),2008,33(5):456-460.
[15]SPIGSET O,MOLDEN E.Cytochrome P450 3A4-the most important arena for drug interactions in the body[J].Tidsskr Nor Laegeforen,2008,128(24):2832-2835.