【作者】 高利宏；

【导师】 曹佳；

【作者基本信息】 第三军医大学 ， 卫生毒理学， 2005， 博士

【摘要】 药物不良反应是长期以来危害人体健康的一个医学难题,当前各国由药物不良反应所引发的疾病都呈上升趋势,同时它也是众多新药开发和临床运用的难题和障碍,对药物毒性机制进行深入研究有重要的医学和社会意义,但传统的药物毒理学研究缺乏在分子水平上进行综合分析的手段,使药物毒理研究与发展相对滞后。毒理基因组学为药物毒理研究引入了新的研究理论和技术手段,基因芯片等高通量基因组技术手段可以同时观察药物毒性作用引发的数以千计的基因的表达变化模式,而这种基因表达谱上所显现的毒性终点往往具有较好的敏感性和特异性,有助于对毒性机制的阐明。目前此方面工作正在广泛开展并取得许多成果。肝脏是人体药物代谢的主要器官,药物引发的肝毒性损伤占临床药物不良反应的相当比例,因此众多研究均从药物肝毒性动物模型入手。本课题是以本室与香港城市大学联合构建的小鼠毒理基因芯片为技术平台,建立四种具肝毒性效应的临床药物(红霉素、四环素、环磷酰胺、异环磷酰胺)的Balb/c小鼠肝毒性模型,对比研究各药物作用后不同时相点和剂量点小鼠肝脏基因表达谱变化,为在分子水平上探索药物肝脏毒性作用机制积累资料,同时验证本实验室与香港城市大学联合构建的小鼠毒理基因芯片用于药物毒性评价的可行性。本研究分为三部分进行:第一部分,建立四种药物致Balb/c小鼠肝毒性损伤模型;第二部分,设计制作小鼠毒理基因芯片,建立相应芯片检测流程,并进行相关质控分析和可靠性验证;第三部分,利用小鼠毒理基因芯片观察各实验组小鼠肝脏基因表达谱变化,并结合聚类分析及生物信息数据库检索等手段对数据进行初步分析,探讨药物致肝脏毒性的作用机制。主要的结果如下:1.本研究建立了红霉素、四环素、环磷酰胺和异环磷酰胺四种药物的Balb/c小鼠不同时相和剂量的肝毒性作用模型,体重指标统计发现多个药物组动物平均体重下降,提示动物机体能量代谢障碍;各组动物血浆肝功生化指标和肝脏病理切片观察发现各药物引起肝脏不同程度和类型的毒性损伤。2.挑选与环境化合物致癌、外来化合物/药物代谢及毒性效应相关的1796个基因片段,构建小鼠毒理基因芯片及配套数据库。更多还原

【Abstract】 The adverse drug reaction (ADR) is a difficult medical problem harmed the humanity health since long ago.The diseases caused by ADR in various countries are on the rise at present , furthermore, ADR is a huge barrier for the the new medicine research and clinical exertion.It has important medical and social significance for carrying on further research on toxic mechanism of medicine. Whereas,there are littler synthetical analyse measure on the molecule level in the traditional medicine toxicology, which result the slowly developing in medicine toxicology studying .Toxicogenomics supply a new technological means and research idea for the medicine toxicology. Gene chip ,which is a high flux genome-based technology , can observe simultaneously thousands gene expression model induced by drug toxic effect, and the toxicity endpoints appeared from the gene expression profile generally are more sensitive and differential that in favor of elucidating the toxicity mechanism.This research work has extensively launched and make a lot of achievements in this respect at present. The liver is a main organ of the human drug metabolism, and the liver damages induced by drug toxicity are high percentage in clinic ADR.Therefore many correlative studying are focus on drug hepatotoxicity.The objective of this experimental study is comparely studying the gene expression profile at different time phases and dosages after drug treated, which based on the balb/c mice hepatotoxic model caused by erythromycin lactobionate, tetracycline hydrochloride, cyclophosphamide, ifosfamide ,and on the mouse toxicology gene chip , accumulating data for the drug hepatotoxicity mechanism on molecule level, and validating the feasibility of the mouse toxicology gene chip applying on drug toxicity evaluation. The experimental study was composed of three parts:⑴Establishing the balb/c mice hepatotoxic model caused by erythromycin lactobionate, tetracycline hydrochloride, cyclophosphamide and ifosfamide.⑵Establishing and evaluating the technique of detection for the mouse更多还原