【作者】 林艳萍；

【导师】 刘昌孝；

【作者基本信息】 天津大学 ， 制药工程， 2009， 博士

【摘要】 肝脏是机体代谢的枢纽,承担了大部分的合成、分解、转化等代谢过程,其中某些酶系和功能是肝脏所特有的,因此,系统研究肝脏损伤后的代谢组变化非常重要。代谢组学主要研究细胞调节过程的终端产物,研究结果能够定量、定性的反映生物体对基因或环境变化的复杂响应,为了解生命奥秘提供了一个新途径。本文将液相色谱-质谱联用技术和化学计量学相结合,整合建立了一个代谢组学的研究平台,并将该平台应用于研究肝损伤的标志物。给Wistar大鼠口服了经典的肝毒性化合物四氯化碳,建立肝毒性模型。在给药前一天和给药后六天内,连续收集了实验动物的尿液和胆汁。采用具有较广泛样品适应性的液相色谱作为分离手段,以高灵敏度、高选择性的质谱作为检测仪器,分析复杂生物样品。在大鼠尿液中找出的病理标志物表明,四氯化碳减缓了大鼠的能量代谢,引起急性肝、肾损伤。大鼠的胆汁排泄也因肝功能受损而减缓,并发现胆汁中卵磷脂的含量有所增加。用代谢组学的研究方法对乙肝病人和正常病人的血清样品进行比对,发现病人血清中的一些组分例如肌酸苷、苏氨酸、酪氨酸和鹅脱氧胆酰甘氨酸等会因为受到病毒感染而浓度升高。实验同时证实临床案例存在很大的个体差异,增加了代谢组学应用于疾病诊断的复杂性。本文通过建立代谢组学平台比较动物模型和人肝损伤的标志物后发现,如果肝实质细胞受损,体内的能量代谢减缓,胆汁酸合成也受影响。本文是代谢组学在肝毒性研究和临床诊断方面应用的另一次尝试。更多还原

【Abstract】 Liver is the key organ for metabolism, because it is responsible for most of the anabolism, catabolism and transforlation in vivo. Some metabolic enzymes are only found in liver, so it’s essential to investigate metabolic mechanisms of liver for the better perspective of hepatic diseases. Metabonomics broadly aims to measure the global, dynamic metabolic response of living systems to biological stimuli or genetic manipulation. It focuses on tracing systemic changes according to time in complex multicellular systems. In this dissertation, there is an integrated metabonomic platform established basing on liquid chromatography-mass spectrometry technique and chemometric methods for the profiling of hepatic injury biomarkers.Urine and bile pre- and post dose are collected for seven-day long from a hepatotoxic model of Wistar rat induced by a single oral dosage of carbon tetrachloride (CCl4). The fluctuations of these urinary components characterize the disfunction of Kerbs Circle as well as an energy decline, proving an acute hepatotoxicity and nephrotoxicity right after the oral exposure of CCl4. Meanwhile, the excretion of bile is slowed down and the concentrations of phosphatidylcholine in bile are elevated.Serums of hepatitis B virus infected people and healthy people are compared through the metabonomic platform. Some components in serum including creatinine, L-Threonine, L-Tyrosine and glycohenodeoxycholic acid or its isomer glycodeoxycholic acid are upregulated after hepatitis B infection. The results also reveal individual variations of clinical samples, which increase challenges of metabonomic investigation.After comparing the hepatic injury biomarkers of hepatotoxic animal model and hepatitis B infected human, we find out that the energy circle is slowed down and biosythesis of bile acid is badly affected if the hepatic cell is injured. The current study is another approach of metabonomics applied in toxicology and clinical diagnosis.更多还原