NMR STUDIES OF CELLULAR METABOLISM

人工肝的发展代表了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仅取得了适度的成功，系统寿命相对较短，并且显示出药物代谢酶如细胞色素P450的表达较差。然而，干细胞技术发展的最新进展使这一目标更加接近现实。我们最近与北卡罗来纳州大学的Lola Reid教授和Jeffrey Macdonald博士合作，利用NMR作为非侵入性工具，评估几种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肝祖细胞进行负分选，从而富集肝祖细胞群体。我们最近也已经能够证明的可行性，NMR监测氟化异生质代谢，以及一些其他的代谢参数在市售的生物反应器。利用模拟人类肝小叶的微型生物反应器进行的初步NMR研究也已完成。我们最近还使用NMR光谱学来了解硼酸盐的生理学和毒理学。这些研究涉及硼酸盐，丝氨酸和γ-谷氨酰转肽酶形成的三元复合物的分析。- 人工肝;生物反应器;核磁共振;丙酸盐;细菌细胞壁;硼酸盐;氟代外源性物质

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