【作者】 倪韶青；

【导师】 曾苏； 赵正言；

【作者基本信息】 浙江大学 ， 药物分析学， 2008， 博士

【摘要】 目的建立CsA治疗再生障碍性贫血（再障）儿童的群体药动学模型,考察其群体药物动力学特征,为临床调整个体用药提供方法。方法利用前瞻性调查分析方法,收集浙江大学医学院附属儿童医院222例门诊再障患儿1611个全血稳态谷浓度样本,用NONMEM程序建立群体药物动力学模型,定量考察年龄、身高、体重、BSA、GFR、合并用药强的松、司坦唑醇等固定效应对药物动力学参数的影响。结果CsA稳态谷浓度药物动力学参数能用米-曼氏模型进行较好的拟合,用建模组获得的群体参数在验证组中有较理想的拟合优度。模型中最终保留了年龄、体表面积（BSA）、肾小球滤过率（GFR）、和司坦唑醇合并用药4个固定效应,米-曼氏模型参数V_m和K_m为206（mg/d）、39.9（μg/L）,V_m和K_m的个体间变异分别是28.0%和67.7%,个体自身的变异为29.2%。用全数据集模拟的最终模型为:DR=[206+9.64×（AGE-8.26）+225×（BSA-1.06）]×Exp（η₁）×Css/（{[39.9+3.61×（GFR-1565）]×（1+0.868×STZ）}×Exp（η₂）+Css）+ε结论环孢素治疗儿童再障具有明显的年龄依赖性的药动学变化特征。用本研究所得到的群体药物动力学模型结合Bayesian反馈法可以为临床儿童血液病患儿CsA的个体化用药提供方法。目的:初步研究不同出生体重新生大鼠CYP3A1/2、PXR、CAR和HNF4α表达差异及其影响CYP3A1/2的调控因素。方法:怀孕第2天的SD孕鼠,随机分配入自由进食组（nourished rats;NR）和营养限制组（undernourished rats,UR）,NR组母鼠所生仔鼠,且体重在5.2～7.2g之间的为正常出生体重组（normal birth weight group,NBW,n=15）;UR组母鼠所生仔鼠且体重＜5.2g为低出生体重组（low birth weight group,LBW,n=15）。NR组母鼠所生仔鼠且体重＞7.2g的新生大鼠为HBW（High birth weight group,HBW,n=15）组。Real-time RT-PCR方法测定CYP3A1/2、PXR、CAR和HNF4α肝组织mRNA表达。免疫组织化学方法检测CYP3A1/2,CAR和HNF4α蛋白表达和组织定位,对染色强度和阳性细胞数计分后评价蛋白表达差异。结果低出生体重显著影响了CYP3A2和CAR的mRNA表达以及CYP3A1和CAR的蛋白表达。而高出生体重显著影响了CYP3A2和HNF4α的mRNA水平,以及CYP3A1和HNF4α的蛋白表达水平。CYP3A1的mRNA与CYP3A2和HNF4α的mRNA的表达具有相关性。结论出生体重对CYP3A1/2以及核受体的表达具有不同程度的影响。提示,需注意不同出生体重新生儿对药物的反应性差异。目的研究不同发育阶段、高脂饮食状态下CYP3A1/2以及核受体的表达变化规律。探讨影响发育过程中CYP3A1/2表达调控的重要因素。方法LBW模型建立、mRNA和蛋白测定方法同本章第一节。模型分组:NBW-常规饲料组（normal birth weight-normal diet group,NN组）NBW-高脂饲料组（normal birth weight-high lipid and high energy group,NH组）LBW-常规饲料组（low birth weight-normal diet group,LN组）LBW-高脂饲料组（loW birth weight-high lipid and high energy group,LH组）以上各组大鼠分别在生后3、7、14、21、28、56和84天收集肝脏（n=6-8/组）。结果CYP3A1/2以及核蛋白PXR、CAR、HNF4α的表达具有年龄依赖性。低出生体重可以改变代谢酶和核受体表达并且一直持续到成年期,其中对PXR/CAR的改变尤为明显,而且在很大程度上与CYP3A1/2表达相关。高脂饮食可以改变代谢酶和核受体的表达,其中HNF4α基因表达的改变最为显著。结论发育过程中CYP3A以及相关核受体表达的改变和相互作用,会导致机体对物质代谢的变化,影响治疗药物的安全性和药物相互作用,临床上需关注低出生体重和肥胖患儿的合理用药。更多还原

【Abstract】 ObjectivesThe aim of this study was to develop a population pharmacokinetic model of cyclosporin（CsA） in pediatric aplastic anemia patients,thereby optimizing its therapeutic effects in clinical trials.MethodsWe have prospectively collected 1611 routine samples of average steady-state CsA trough concentration in 222 pediatric patients with hematologic disease in a Chinese population.These data were analyzed using the nonlinear mixed-model program（NONMEM）.Based on the influence of numerous tested covariates（i.e.,age, height,weight,BSA,GFR）,a population pharmacokinetic model of CsA was developed and further validated by data splitting.ResultsAfter oral administration,the whole blood steady-state trough concentration of CsA could be well described by a Michaelis-Menten model,in which four covariates （i.e.,age,BSA,GFR,and stanozolol） were included.Furthermore,the Michaelis-Menten constants,Vm and Km with interindividual variation in parenthesis, were estimated to be 206 mg/d（28%） and 39.9μg/L（67.7%）,respectively.The intraindividual variation of V_m and K_m was 29.2%.The final model was described as: DR=（[206+9.64×（AGE-8.26）+22.5×（BSA-1.06）]×Exp（η₁）×Css）/({[39.9+3.6l×（GFR-1565]×（1+0.868×STZ）}×Exp（η₂）+Css）+εConclusionsIn conclusion,pharrnacokinetic parameters of CsA in pediatric aplastic anemia patients may be largely influenced by age,BSA,GFR and Stanzolol.We propose that our CsA population pharrnaeokinetie model,combined with the Bayesian forecasting method,may optimize the dosage of CsA in clinical trials. ObjectivesThe aim of this study was to investigate the effects of birth weight on the expression and regulation of CYP3A1/2 as well as its nuclear receptors,including PXR, CAR,and HNF4a in newborn rats.MethodsPregnant rats were randomly divided into two groups:nourished and under-nourished.The offspring of nourished rats with birth weight between 5.2～7.2g were defined as normal-birth-weight group（NBW,n=15）,while those of undernourished rats with birth weight under 5.2g were defined as low-birth-weight group（LBW,n=15）.Additionally,the offspring of nourished rats with birth weight above 7.2g were defined as high-birth-weight group（HBW,n=15）.Hepatic mRNA expression of CYP3A1,PXR.,CAR and HNF4αwas detected by quantitative real-time RT-PCR.Immunohistochemistry was performed to investigate the corresponding protein expression and localization.Protein expression was evaluated by using a score corresponding to the sum of both staining intensity and percentage of positive hepatocytes.ResultsLow birth weight could significantly affect mRNA expression of CYP3A2 and CAR,and protein expression of CYP3A1 and CAR.In contrast,the high birth weight could significant affect the mRNA levels of CYP3A2 and HNF4α,and also the protein levels of CYP3A1 and HNF4a expression.CYP3A1 mRNA expression correlated with that of CYP3A2 and HNF4a.ConclusionsIn conclusion,birth weight could influence the production of several drug metabolism enzymes（i.e.,CYP3A1/2,and its nuclear receptors） in newborn rats, which should be taken into consideration when delivering medication to LBW and HBW infants in humans. ObjectivesIn this study,we have aimed to investigate the effects of low birth weight and high-lipid diet on the expression and regulation of CYP3A1/2 as well as its nuclear receptors,including PXR,CAR,and HNF4α,in female rats during development.MethodsThe LBW animal model was established as mentioned above in Part One.Gene expression analysis（mRNA and protein levels） was performed as described previously. According to different birth weight and diet feeding,rats were divided into 4 groups as following:1.Normal birth weight-normal diet group,NN-group2.Normal birth weight-high lipid and high energy group,NH-group3.Low birth weight-normal diet group,LN-group4.Low birth weight-high lipid and high energy group,LH-groupLivers were isolated from each group,and subsequently used for gene expression study （i.e.,mRNA and protein levels） at 3,7,14,21,28,56,84 days after birth（n=6-8 per group）.ResultsThe hepatic expression of CYP3A1/2 and its nuclear receptors（i.e.,PXR,CAR, and HNF4α） was found to be age’dependent in rats.The effects of low birth weight on the expression of CYP3A1/2,PXR,CAR and HNF4αcan persist after birth.In particular,the expression of PXR and CAR was dramatically altered under the LBW situation,which was correlated with the expression of CYP3A1/2.Furthermore,the expression of CYP3A1/2 and their nuclear receptors,especially the HNF4α,was largely influenced by the high-lipid diet feeding in rats.ConclusionIn conclusion,the hepatic expression and modulation of CYP3A1/2 and its nuclear receptors（i.e.,PXR,CAR,and HNF4α） were influenced by both birth weight and high-lipid diet feeding in developing rats.We propose that the alteration and interaction of CYP3A and its nuclear receptors have potential clinical implications for optimal medication in low birth weigh and fat children.更多还原