梁淼;马增春;易剑峰;王宇光;谭洪玲;肖成荣;梁乾德;汤响林;李桦;沈国林;高月;
LIANG Miao;MA Zeng-chun;YI Jian-feng;WANG Yu-guang;TAN Hong-ling;XIAO Cheng-rong;LIANG Qian-de;TANG Xiang-lin;LI Hua;SHEN Guo-lin;GAO Yue;School of Pharmacy,Guangxi Medical University;Research Institute of Radiation Medicine,Academy of Military Medical Science;Research Institute of Toxicology and Pharmacology,Academy of Military Medical Science;

• 1:广西医科大学药学院
• 2:军事医学科学院放射与辐射医学研究所
• 3:军事医学科学院毒物药物研究所

摘要(Abstract)：

研究四物汤复方及其配伍对大鼠肝脏P450酶活性及其mRNA表达水平的影响。大鼠口服灌胃四物汤及其配伍水煎液2周后,处死,用生理盐水灌流肝脏,制备肝微粒体,采用混合探针与肝微粒体体外孵育法考察四物汤对大鼠肝脏细胞色素P450酶的影响。利用实时定量聚合酶链式反应(Q-PCR)检测四物汤对大鼠肝脏细胞色素P450 mRNA表达的影响。四物汤复方组CYP1A2酶活性与对照组相比升高(P<0.05),熟地白芍、当归白芍、当归川芎以及白芍川芎组的CYP1A2和CYP2C19的酶活性与对照组相比降低(P<0.05),复方组、单药组、2味药配伍组的CYP2B6的酶活性与对照组相比降低(P<0.05)。复方组对CYP1A2的mRNA表达具有上调作用(P<0.05),4味单药对CYP2B1 mRNA表达具有下调作用的(P<0.05)。四物汤复方对大鼠肝脏CYP1A2的酶活性具有诱导作用,熟地和当归组与对照组相比酶活性相当,但与白芍合用后显著下调了CYP1A2的酶活性。复方及其配伍对CYP2B6的酶活性具有抑制作用,其中当归对酶活性下调作用较强。复方对CYP1A2以及4味单药对CYP2B1的mRNA表达的影响与酶活性水平相平行。
To study the effect of Siwu decoction(SWD)compound and its combined administration on hepatic P450 enzymatic activity and mRNA expression in rats. Rats were orally administered with SWD and water decoction combined with other medicines for two weeks,and then sacrificed. Their livers were perfused with normal saline to prepare liver micrisomes. Mixed probe and liver microsome in vitro incubation method were adopted to detect the effect of SWD on hepatic cytochrome P450. The real-time quantitative polymerase chain reaction(Q-PCR)was used to detect the effect of SWD on the expression of hepatic cytochrome P450. Compared with the control group,the SWD compound group showed higher CYP1A2 enzymatic activity( P < 0. 05); Rehmanniae-paeoniae,angelicaepaeoniae,angelicae-rhizome,paeoniae-rhizome groups had lower CYP1A2 and CYP2C19 enzymatic activities( P < 0. 05); And the compound group,the single component group and the combination group showed lower CYP2B6 enzymatic activities( P < 0. 05). The compound could up-regulated the mRNA expression of CYP2B1( P < 0. 05); And the four single components could down-regulated the mRNA expression of CYP2B1( P < 0. 05). SWD compound had the effect in inducing CYP1A2 enzymatic activity. The rehmanniaepaeoniae group and the angelicae-paeoniae group had identical enzymatic activity with the control group,but significant down-regulation in CYP1A2 enzymatic activity after being combined with paeoniae. The compound and its combined administration showed the inhibitory effect on CYP2B6 enzymatic activity,particularly being combined with angelicae. The compound showed identical effect with the four single components in terms of CYP1A2 mRNA expression and enzymatic activity.

关键词(KeyWords)： 四物汤;肝微粒体;细胞色素P450酶
Siwu decoction;liver microsome;cytochrome CYP450 enzyme

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金重点项目(81130067);国家自然科学基金面上项目(81073028,81274127)

作者(Authors): 梁淼;马增春;易剑峰;王宇光;谭洪玲;肖成荣;梁乾德;汤响林;李桦;沈国林;高月;
LIANG Miao;MA Zeng-chun;YI Jian-feng;WANG Yu-guang;TAN Hong-ling;XIAO Cheng-rong;LIANG Qian-de;TANG Xiang-lin;LI Hua;SHEN Guo-lin;GAO Yue;School of Pharmacy,Guangxi Medical University;Research Institute of Radiation Medicine,Academy of Military Medical Science;Research Institute of Toxicology and Pharmacology,Academy of Military Medical Science;

参考文献(References)：

• [1]Elizabeth Schneider,Douglass Clark.Cytochrome P450(CYP)enzymes and the development of CYP biosensors[J].Biosens Bioelectron,2012(13):1.
• [2]Michael Sinz,Gillian Wallace,Jasminder Sahi.Current industrial practices in assessing CYP450 enzyme induction:preclinical and clinical[J].AAPS J,2008,10(2):391.
• [3]王宝莲,盛莉,李燕.混合探针底物法测定五味子策划给你分对大鼠肝脏细胞色素P450酶同工酶的影响[J].药学学报,2011,46(8):922.
• [4]David R Nelson.Cytochromr P450 and the individuality of species[J].Arch Biochem Biophys,1999,369(1):1.
• [5]李晓宇,刘皋林.CYP450酶特性及其应用研究进展[J].中国临床药理学与治疗学,2008,13(8):942.
• [6]Blakey G E,Lockt on J A,Perrett J,et al.Pharmacokinetic and pharmacodynamic assessment of a five-probe metabolic cocktail for CYPs 1A2,3A4,2C9,2D6 and 2E1[J].Br J Clin Pharmacol,2003,57:162169.
• [7]梁乾德,王升启,高月.四物汤基础研究进展[J].天津中医药,2003,20(5):83.
• [8]刘明,高月,肖瑞,等.中药组方原则“君臣佐使”的模糊数学量化描述[J].药学学报,2009,44(1):38.
• [9]孙严彤,郭颖杰,张梦亮,等.新型β2受体激动剂川丁特罗对HepG2细胞中细胞色素P450 mRNA表达的影响[J].吉林大学学报,2010,36(5):862.
• [10]Yong-Long Han,Dan Li,Bin Ren,et al.Evaluation of impact of Herba Erigerontis injection,a Chinese herbal prescription,on rat hepatic cytochrome P450 enzymes by cocktail probe drugs[J].J Ethnopharmacol,2012,139:104.
• [11]Heeyeon Kim,Hyun Kyu Choi,Tae Cheon Jeong,et al.Selective inhibitory effects of mollugin on CYP1A2 in human liver microsomes[J].Food Chem Toxicol,2012(51):33.
• [12]Ernest Hodgson,Randy L Rose.The importance of cytochrome P450 2B6 in the human metabolism of envirmental chemicals[J].Pharmacol Ther,2007,113:420.
• [13]Penelope M Y Or,Francis F Y Lam,Y W Kwan,et al.Effects of Radix Astragali and Radix Rehmanniae,the components of an anti-diabetic foot ulcer herbal formula,on metabolism of model CYP1A2,CYP2C9,CYP2D6,CYP2E1 and CYP3A4 probe substrates in pooled human liver microsomes and specic CYP isoforms[J].Phytomedicine,2012,19:535.
• [14]陈立兵,葛卫红,林洁,等.细胞色素P450 mRNA表达研究进展[J].医学研究杂志,2007,36(7):7.