【作者】 杨海峰；

【导师】 江善祥；

【作者基本信息】 南京农业大学 ， 基础兽医学， 2006， 硕士

【摘要】 本文采用体外微粒体酶生化测定和体内探针药物代谢法评价了鸡肝微粒体细胞色素P450酶系活性，探讨了苯巴比妥和地塞米松对鸡肝微粒体CYP450的诱导作用。 1 鸡肝微粒体细胞色素P450酶系活性检测方法的建立 12只60日龄依莎公鸡颈静脉放血处死，应用差速离心法提取肝微粒体，Bradford法测定肝微粒体蛋白含量，紫外分光光度法测定CYP450酶系活性。结果显示：肝微粒体蛋白含量为11.32±1.43mg·g^-1，细胞色素P450含量为0.71±0.08nmol·mg^-1，NADPH-细胞色素C还原酶活力为3.01±0.26nmol·mg^-1·min^-1，氨基比林-N-脱甲基酶（AND）活力为2.36±0.18nmol·mg^-1·min^-1，红霉素-N-脱甲基酶（ERND）活力为1.21±0.11nmol·mg^-1·min^-1，苯胺-4-羟化酶（AH）活力为0.25±0.03nmol·mg^-1·min^-1。结果表明本实验所建立的鸡肝微粒体细胞色素P450活性的检测方法灵敏可靠，可用于CYP450的系统研究。 2 探针药物安替比林静注在鸡体内的药动学研究 健康鸡12只，一次静脉注射CYP450探针药物安替比林20mg·kg^-1，反相高效液相色谱法（RP-HPLC）法测定血浆中安替比林的浓度，进行药代动力学研究。结果显示：血浆中安替比林在0.1～30μg·mL^-1浓度范围内，药物峰面积与浓度线性关系良好（r=0.9995），最低检测限为0.1μg·mL^-1，低、中、高浓度(0.5、5.0、20μg·mL^-1)的血浆标准品的相对回收率分别为93.44％、95.88％、97.21％，日内和日间变异系数小于5％。安替比林静注后药-时数据符合二室开放模型，主要药动学参数为：CL=0.041±0.01L·kg^-1·min^-1，T_1／2β=29.35±6.57min，AUC=503.25±92.34(μg·mL^-1)·min。结果表明：本实验所建立的安替比林血药浓度测定的RP-HPLC法简便、快速、灵敏、准确、可靠，可用于安替比林消除动力学模型的研究。安替比林静注后在鸡体内分布和消除均较快，所得药动学参数可为探针药物法评价鸡CYP450活性提供理论依据。 3 苯巴比妥和地塞米松对鸡细胞色素P450的诱导作用研究 采用体外肝微粒体酶生化测定和体内探针药物（安替比林）代谢的方法，研究苯巴比妥（PB）和地塞米松（DEX）对鸡细胞色素P450（CYP450）有无诱导作用。将更多还原

【Abstract】 The biochemical determinations of hepatic microsomal enzymes in vitro and the metablism of a probe drug in vivo were conducted to evaluate the activities of microsomal cytochrome P450 enzymes in chickens. Induction of Phenobarbital and dexamethasone on hepatic microsomal cytochrome P450 in chickens was investigated in this paper. The results were summarized as follows:1 Study on methodology for the determination of activities of hepatic microsomal cytochrome P450 enzymes in chickensTwelve 60-day-old cocks were sacrificed by cutting off jugular vein. Liver microsome was distilled by different velocity centrifugation. The Bradford method was used to measure the concentration of protein in hepatic microsome. The activities of Cytochrome P450 were measured by spectrophotometer analysis. The results showed that the concertration of protein in hepatic microsome was 11.32±1.43 mg·g^-1, the content of cytochrome P450 was 0.71±0.08 nmol·mg^-1, NADPH -Cytochrome C reductase activity was 3.01±0.26 nmol·mg^-1·min^-1, aminopyrine-N-demethylase activity was 2.36±0.18 nmol·mg^-1·min^-1, erythromycin-N-demethylase activity was 1.21±0.11 nmol·mg^-1 ·min^-1, and aniline-4-hydroxylase activity was 0.25±0.03nmol·mg^-1·min^-1. The results suggested that the established methodology for determing the content and activities of hepatic microsomal cytochrome P450 enzymes in chickens was sensitive and credible.2 Study on pharmacokinetics of antipyrine probe after intravenous administration in chickensPharmacokinetics of antipyrine probe was studied in twelve healthy chickens after intravenous administration at a single dose of 20 mg·kg^-1. Antipyrine concentration in plasma was determined by a reversed phase high performance liquid chromatography method. The results showed that excellent line relationship was obtained in the range of 0.1 to 30μg·mL^-1 with a correlation coefficient of 0.9995. The lowest detection limit of antipyrine was 0.1μg·mL^-1. The mean recovery rate at the quantitative concentrations of 0.5、1.0、20μg·mL^-1 was 93.44%, 95.88% and 97.21%, respectively. The intra-day and inter-day RSD were both less than 5%. The concentration-time data of antipyrine were fitted to a two-compartment open model. The main parmacokinetic parameters were as更多还原