NMR STUDIES OF CELLULAR METABOLISM

人工肝的开发代表着毒理学和医学的长期目标。利用各种生物反应器模型的尝试只取得了一定的成功，系统的寿命相对较短，并且显示出药物代谢酶的表达较差，如细胞色素P450。然而，干细胞技术发展的最新进展使这一目标变得更加接近现实。我们最近一直在与北卡罗来纳大学的Lola Reid教授和Jeffrey Macdonald博士合作，利用核磁共振作为一种非侵入性工具来评估几个肝脏生物反应器模型系统中的各种代谢和流动参数。Reids博士团队最近开发了一种抗体筛选方法，该方法将用于通过荧光激活细胞分选对人类肝祖细胞进行负向分选，从而使肝祖细胞群体得以丰富。我们最近还证明了在商业上可用的生物反应器中对氟化异种生物代谢以及其他一些代谢参数进行核磁共振监测的可行性。利用模拟人类肝脏小叶的微型生物反应器也进行了初步的核磁共振研究。我们最近还使用核磁共振波谱来了解硼酸盐的生理和毒理学。这些研究包括分析由硼酸盐、丝氨酸和伽马谷氨酰转肽酶形成的三元复合体。人工肝；生物反应器；核磁共振；丙酸；细菌细胞壁；硼酸盐；含氟异物

The development of an artificial liver represents a long standing goal of toxicology as well as medicine. Attempts to utilize various bioreactor models have had only moderate success, with systems being relatively short lived and showing poor expression of drug metabolizing enzymes such as cytochrome P450. However, recent advances in the development of stem cell technology have brought this goal considerably closer to reality. We have recently been working with Prof. Lola Reid and Dr. Jeffrey Macdonald at the University of North Carolina to utilize NMR as a non-invasive tool for assessing various metabolic and flow parameters in several hepatic bioreactor model systems. Dr. Reids group has recently developed an antibody screening method that will be used to negatively sort human hepatic progenitors by fluorescent activated cell sorting, allowing enrichment of hepatic progenitor populations. We also recently have been able to demonstrate the feasibility of NMR monitoring of fluorinated xenobiotic metabolism, as well as a number of other metabolic parameters in a commercially available bioreactor. Preliminary NMR studies utilizing a miniature bioreactor that mimics a human liver lobule have also been performed.We have also recently used NMR spectroscopy to understand the physiology and toxicology of borate. These studies involve the analysis of the ternary complex formed from borate, serine, and the enzyme gamma glutamyl transpeptidase. - artificial liver; bioreactor; NMR; propionate; bacterial cell wall; borate; fluorinated xenobiotics