【作者】 刘文辉；

【导师】 周宏灏；

【作者基本信息】 中南大学 ， 药理学， 2009， 硕士

【摘要】 背景:姜黄素（curcumin）是从姜科植物Curcuma longa的根茎姜黄中提取的一种植物多酚。姜黄素的药理作用十分广泛,包挂抗氧化、抗肿瘤、抗炎、清除自由基、抗微生物以及对心血管系统、消化系统的作用等。CYP2C9主要存在于肝脏中,占肝微粒体P450蛋白总量的20%,仅次于CYP3A。CYP2C9能羟化代谢许多不同性质的药物,主要是酸性底物。据统计,目前约有16%的的临床药物由CYP2C9负责代谢,并在前致癌物、前毒物和致突变剂的活化中也起到一定作用。CYP2C9具有遗传多态性,其中CYP2C9*3突变为亚洲人中主要的与酶活性相关的突变位点。近年研究,发现姜黄素在体外能显著抑制CYP2C9的酶活性,但未见相关体内实验的报道。目的:研究姜黄素对人体内CYP2C9酶活性的影响,以及姜黄素对CYP2C9酶活性的影响作用与CYP2C9基因型之间的关系。方法:体内实验以甲苯磺丁脲为探针药研究姜黄素对人体内CYP2C9活性的影响作用。采用聚合酶链式反应-限制性片断长度多态性（PCR-RFLP）的方法在大样本人群中进行基因型分析,挑选出CYP2C9*1*1和CYP2C9*1*3各7人参加临床试验。临床实验按照两阶段交叉方案进行,两次试验期间有14天的洗脱期。第一阶段按照基因型分成的第一组受试者随机口服临床剂量的姜黄素（1000mg/d,共14天）,第二组不给予处理,第15天给所有受试者均口服500mg甲苯磺丁脲。抽取服药后0,0.25,0.5,0.75,1,1.5,2,2.5,3,4,6,8,10,12,24小时的血样,并收集0-12h的尿样,经过14天洗脱期后交叉重复试验,离心分离血浆,置-40℃低温冰箱冷冻保存至测定。以0-12小时尿液代谢产物羟基甲苯磺丁脲排出量与母药甲苯磺丁脲0-12小时AUC的比值作为CYP2C9的活性指标。采用HPLC法测定甲苯磺丁脲及其代谢产物羟基甲苯磺丁脲的血药浓度。结果:单服甲苯磺丁脲时,CYP2C9*3杂合子与野生型比较,T_1/2显著增加（7.52±4.09 VS 14.78±3.49,P＜0.05）,T_max显著增加（2.00±0.58VS3.79±1.63,P＜0.05）,AUC_（0-24h）显著增加（2239.01±578.63 VS 3352.40±624.37,P＜0.05）,AUC_（0-∞）显著增加（2596.08±925.81 VS 4911.75±765.32,P＜0.05）,尿_{羟甲（0-12h）}/血AUC_（0-12h）显著降低（24.93±10.35 VS 11.7±4.27,P＜0.05）。经姜黄素处理后,T_max显著减小（2.89±1.50 VS 1.75±0.85,P＜0.05）,AUC_（0-24h）显著降低（2820.33±852.69μg.h/ml 2278.05±601.35μg.h/ml,P＜0.05）,AUC_（0-∞）显著降低（3482.65±1376.30μg.h/mlVS 2824.12±1059.92μg.h/m1,P＜0.05）,尿羟甲_（0-12h）/血AUC_（0-12h）显著升高（13.26±9.57ml/h VS 16.38±11.76 ml/h,P＜0.05）。经姜黄素处理后,CYP2C9野生型的AUC_（0-24h）显著降低（2239.01±578.63μg.h/ml 1893.75±414.17μg.h/ml,P＜0.05）,AUC_（0-∞）显著降低（2596.08±925.81μg.h/ml 2134.62±543.23μg.h/ml,P＜0.05）,尿_{羟甲（0-12h）}/血_{AUC（0-12h）}值显著升高（24.93±10.35ml/h VS 33.19±17.18ml/h,P＜0.05）。CYP2C9突变型杂合子T_max显著减小（3.79±1.63 VS 2.07±0.89,P＜0.05）,AUC_（0-24h）显著降低（3352.40±624.37μg.h/ml VS 2645.2±511.73μg.h/ml,P＜0.05）。结论:1.CYP2C9.3突变影响甲苯磺丁脲的药代动力学参数;2.姜黄素能显著促进甲苯磺丁脲的代谢,使代谢产物的排出增加,表明姜黄素对人体内CYP2C9的活性有诱导作用;3.CYP2C9的基因多态性与姜黄素对甲苯磺丁脲的药代动力学的影响没有相关性。更多还原

【Abstract】 Background: Curcumin is a plant polyphenol witch is extracted from the Zingiberaceae rhizome of Curcuma longa. Curcumin has a wide range of pharmacological effects, including linked antioxidant, anti-tumor, anti-inflammatory, free radical scavenging, anti-microbial as well as the cardiovascular system, the role of the digestive system. It has become increasingly clarified CYP2C9 is a very important drug metabolizing CYP isoform in human liver. CYP2C9 is principally responsible for the metabolism of a number of clinically important durgs.It is also involved in the metabolic activation of certain pro-drugs and certain pro-carcinogens. CYP2C9 has polymorphisms, of which CYP2C9*3 mutation are major alleles related to alteration of enzyme activity in Asians. Currently, several researchers has found that curcumin is able to increase the activity of CYP2C9 in vitro. However, there is no study about the effect of curcumin on CYP2C9 in vivo that has been reported.Objective: To investigate the effect of curcumin on CYP2C9 activity in vivo.At the same time to explore the relationship between the effect of curcumin on CYP29 and CYP2C9 genotype.Methods: An open, randomize, 2-period study was performed in 14 healthy male volunteers. Tolbutamide （a CYP2C9 substrate） was used as a probe. Genotypes were determined using the PCR-RFLP assay. Suitable subjects have been selected out, including 7 （CYP2C9*1*1） and 7 （CYPC29*1*3） genotype. 14 healthy volunteers were divided into two groups randomly. Two groups were administered by a single oral 500 mg tolbutamide before and after treatment with curcumin for 14 days （n=7）.5ml blood samples were collected at Oh, 0.25h, 0.5h, 0.75h, 1h, 1.5h, 2h, 2.5h, 3h, 4h, 6h, 8h, 10h, 12h, 24h and 6h to12h urine was collected after the 500mg tolbutaminde tablet.Then the collections of tolbutamide and hydroxytolbutamide in blood an urine were determined by HPLC.CYP2C9 activity was measured as the ratio of metabolites（hydroxytolbutamide） extreted in the 0h to 12h urine to the AUC_1-12h of tolbutamide in blood.Results: After 14 days consecutive administration of curcumin, Tmax significantly decreased （2.89±1.50 VS 1.75±0.85）. AUC_（0-24） shortened significantly（2820.33±852.69μg.h / ml 2278.05±601.35μg.h / ml , P＜0.05）, AUC_（0-∞） shortened significantly (（3482.65±1376.30μg.h / ml VS 2824.12±1059.92μg.h / ml, P＜0.05）. And urine _{hydroxymethyl（0-12h）} / blood _{AUC（0-12h）} significantly increased （13.26±9.57 ml/hVS 16.38±11.76 ml/ h, P＜0.05）, which implied that curcumin induced the activity of CYP2C9. There is no significant relationship between the induction and the CYP2C9 genotypes.Conclusion: In conclusion, curcumin is able to significantly induce the activity of CYP2C9 in vivo, while there is no significantly relationship between the inhibition of curcumin on the activity of CYP2C9 and CYP2C9 genotypes. Therefore, it is necessary to pay great attention when the drug interaction occurs between curcumin and CYP2C9 substrates.更多还原