藏药佐太对药物代谢酶CYP2C11和CYP2D1的活性及蛋白和mRNA表达的影响
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摘要
目的探讨佐太对药物代谢酶CYP2C11和CYP2D1活性,以及蛋白和mRNA表达的影响。方法 140只SD大鼠,雌雄各半,随机分为CYP2C11(给予探针药物苯妥英)和CYP2D1(给予探针药物右美沙芬)两大组,每大组设7个小组,为对照组、小剂量单次给药组、小剂量多次给药组、中剂量单次给药组、中剂量多次给药组、大剂量单次给药组、大剂量多次给药组,每小组10只。采用探针药物法测定大鼠给予佐太后药物代谢酶活性的变化;ELISA法测定大鼠给予佐太后药物代谢酶蛋白表达的变化;实时定量PCR法测定大鼠给予佐太后药物代谢酶mRNA表达的变化。结果与对照组比较,单次给予佐太12.0 mg·kg~(-1)后,CYP2C11的活性显著升高105.3%;单次给予佐太3.8 mg·kg~(-1)后,CYP2D1的活性显著升高193.5%;CYP2D1的蛋白表达显著升高84.3%;CYP2D1的mRNA表达显著升高76.4%,其他组无显著性变化。多次给予佐太1.2、3.8、12.0 mg·kg~(-1)后,CYP2C11的活性分别显著升高436.8%、131.6%、226.3%;CYP2C11的蛋白表达分别显著升高88.1%、38.7%、54.2%;CYP2C11的mRNA表达分别显著升高126.8%、135.2%、100.0%;CYP2D1的活性分别显著升高145.2%、71.0%、58.1%;CYP2D1的蛋白表达分别显著升高84.5%、73.7%、33.4%;CYP2D1的mRNA表达分别显著升高118.2%、67.3%、36.4%,其他组无显著性变化。结论藏药佐太对CYP2C11和CYP2D1的活性,蛋白质及mRNA表达均具有显著升高效果,显示佐太在作为合剂时可诱导肝药酶代谢加快。
Objective To investigate the effects of Tibetan medicine Zuotai on the activity,protein and mRNA expression of CYP2C11 and CYP2D1. Methods 140 SD rats are randomly divided into two groups with half male and female: CYP2C11(treated with probe drug phenytoin) and CYP2D1(treated with probe drug dexamethasone).Each group were divided into seven groups with ten rats which are control group,single low-dose administration group,multiple low-dose,single middle-dose,multiple middle-dose,single high-dose,multiple high-dose. The activity of drug metabolizing enzyme in rats was determined by probe drug method.The protein and mRNA expression of drug metabolizing enzyme in rats were determined by ELISA and real-time quantitative PCR. Results Compared with the reference,the activity of CYP2C11 was significantly increased by 105.3% after treated with 12.0 mg·kg~(-1) Zuotai single administration. The activity of CYP2D1 was significantly increased by 193. 5% after treated with3.8 mg·kg~(-1) Zuotai single administration.The protein and mRNA expression of CYP2D1 was drastically enhanced over 84.3% and 76.4%,and there was no significant change in other groups. After treated with Zuotai multiple administration of 1.2,3.8,12.0 mg·kg~(-1),the activity of CYP2C11 and CYP2D1 show 436.8%,131.6%,226.3% and145.2%,71.0%,58.1% improvement,respectively.The protein and mRNA expression of CYP2C11 were drastically enhanced over 88.1%,38.7%,54.2% and 126.8%,135.2%,100.0%,respectively. The protein and mRNA expression of CYP2D1 were drastically enhanced over 84.5%,73.7%,33.4% and 118.2%,67. 3%,36. 4%,respectively.There was no significant change in other groups.Conclusion Tibetan medicine Zuotai can drastically enhance the activity,protein and mRNA expression of CYP2C11 and CYP2D1,show that Zuotai as a mixture can induce accelerating liver enzyme metabolism.
Objective To investigate the effects of Tibetan medicine Zuotai on the activity,protein and mRNA expression of CYP2C11 and CYP2D1. Methods 140 SD rats are randomly divided into two groups with half male and female: CYP2C11(treated with probe drug phenytoin) and CYP2D1(treated with probe drug dexamethasone).Each group were divided into seven groups with ten rats which are control group,single low-dose administration group,multiple low-dose,single middle-dose,multiple middle-dose,single high-dose,multiple high-dose. The activity of drug metabolizing enzyme in rats was determined by probe drug method.The protein and mRNA expression of drug metabolizing enzyme in rats were determined by ELISA and real-time quantitative PCR. Results Compared with the reference,the activity of CYP2C11 was significantly increased by 105.3% after treated with 12.0 mg·kg~(-1) Zuotai single administration. The activity of CYP2D1 was significantly increased by 193. 5% after treated with3.8 mg·kg~(-1) Zuotai single administration.The protein and mRNA expression of CYP2D1 was drastically enhanced over 84.3% and 76.4%,and there was no significant change in other groups. After treated with Zuotai multiple administration of 1.2,3.8,12.0 mg·kg~(-1),the activity of CYP2C11 and CYP2D1 show 436.8%,131.6%,226.3% and145.2%,71.0%,58.1% improvement,respectively.The protein and mRNA expression of CYP2C11 were drastically enhanced over 88.1%,38.7%,54.2% and 126.8%,135.2%,100.0%,respectively. The protein and mRNA expression of CYP2D1 were drastically enhanced over 84.5%,73.7%,33.4% and 118.2%,67. 3%,36. 4%,respectively.There was no significant change in other groups.Conclusion Tibetan medicine Zuotai can drastically enhance the activity,protein and mRNA expression of CYP2C11 and CYP2D1,show that Zuotai as a mixture can induce accelerating liver enzyme metabolism.
引文
[1]张炜,刘学良,骆桂法,等.佐太等珍宝矿物类藏药材中总砷、总汞及可溶性砷汞含量的对比[J].药物分析杂志,2016,4(36),721-726.
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[9]李向阳,朱俊博,尼玛才让,等.微波消解ICP-AES法同时测定藏药佐太中的4种金属元素[J].中成药,2009,31(9):1472-1473.
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[12]万玛才旦.藏药大月晶丸治疗消化性溃疡症55例临床观察[J].中国民族医药杂志,1997,21(4):27-27.
[13]程体娟,杨军英,朱玲,等.仁青芒觉胶囊的耐缺氧及抗心律失常作用[J].中药药理与临床,2003,19(2):40-41.
[14]桑吉措.藏药仁青常觉对101例消化系统疾病的临床观察[J].中国民族医药杂志,1999,5(3):5-6.
[15]朱洪梅,魏立新,杜玉枝,等.藏药佐太长期给药对小鼠毒性的初步研究[J].时珍国医国药,2013,24(8):2022-2024.
[16]李向阳,冯伟力,马祁生,等.藏药佐太中的汞在大鼠体内吸收和排泄的初步研究[J].时珍国医国药,2010,21(2):290-292.
[17]陈朝军,周昊霏,陆景坤,等.不同产地寒水石中重金属及有害元素的含量研究[J].世界科学技术-中医药现代化,2012,14(3):1678-1682.
[2]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.
[3]杨梦,周雪姣,李向阳.藏药佐太的研究现状与进展[J].药学研究,2016,35(11):663-667.
[4]看召本.透视藏医珍宝类药品中的佐太[J].中国中药杂志,2013,38(10):1621-1623.
[5]黄海波,王奇志,王新为,等.藏药佐太的研究进展[J].中国中药杂志,2013,38(17):2886-2888.
[6]向丽,曾勇,林波,等.藏药“佐太”长期给药对大鼠肾脏潜在氧化损伤的初步研究[J].中药与临床,2015,6(3):39-42.
[7]李岑,王东平,多杰,等.藏药佐太安全性研究及其复方当佐的临床安全观察初探[J].中国中药杂志,2014,39(13):2573-2582.
[8]李向阳,刘永年,李永平,等.藏药佐太对CYP1A2和NAT2的活性及蛋白和mRNA表达的影响[J].药学学报,2014,49(2):267-272.
[9]李向阳,朱俊博,尼玛才让,等.微波消解ICP-AES法同时测定藏药佐太中的4种金属元素[J].中成药,2009,31(9):1472-1473.
[10]Anwar-Mohamed A,Elbekai RH,El-Kadi AO.Regulation of CYP1A1 by heavy metals and consequences for drug metabolism[J].Expert Opin Drug Metab Toxicol,2009,5(5):501-521.
[11]Anwar-Mohamed A,El-Kadi AO.Down-regulation of the carcinogen metabolizing enzyme cytochrome P450 1a1 by vanadium[J].Drug Metab Dispos,2009,36(9):1819-1827.
[12]万玛才旦.藏药大月晶丸治疗消化性溃疡症55例临床观察[J].中国民族医药杂志,1997,21(4):27-27.
[13]程体娟,杨军英,朱玲,等.仁青芒觉胶囊的耐缺氧及抗心律失常作用[J].中药药理与临床,2003,19(2):40-41.
[14]桑吉措.藏药仁青常觉对101例消化系统疾病的临床观察[J].中国民族医药杂志,1999,5(3):5-6.
[15]朱洪梅,魏立新,杜玉枝,等.藏药佐太长期给药对小鼠毒性的初步研究[J].时珍国医国药,2013,24(8):2022-2024.
[16]李向阳,冯伟力,马祁生,等.藏药佐太中的汞在大鼠体内吸收和排泄的初步研究[J].时珍国医国药,2010,21(2):290-292.
[17]陈朝军,周昊霏,陆景坤,等.不同产地寒水石中重金属及有害元素的含量研究[J].世界科学技术-中医药现代化,2012,14(3):1678-1682.