BIOENERGETIC REQUIREMENTS FOR A BIOARTIFICIAL PANCREAS

生物人工胰腺承诺生理控制血糖水平，有效治疗I型糖尿病。鉴于胰岛供应短缺，有必要开发适用于此类装置的替代细胞系。转化细胞是一个可行的选择，尽管它们的生物能量机制可能效率低下。 由于生物能量学在胰岛素分泌中起作用，因此确定对胰岛素分泌至关重要的生物能量因素对于选择合适的线以包含到生物人工胰腺中可能是重要的。这项提案将研究文化的截留转化胰岛素瘤细胞，并确定ATP和回补途径的影响，他们的能力分泌胰岛素，以响应外部葡萄糖刺激。 13C和31P数据的NMR光谱分析将提供能量代谢和胰岛素分泌之间的耦合的非侵入性定量评估，而非NMR测定将导出葡萄糖和氧消耗速率，以及乳酸盐、NH4+和胰岛素产生速率以提供代谢谱。 TCA循环、回补途径和氧化磷酸化的特异性抑制剂的给药将有助于识别启动胰岛素分泌的关键因素。 这些知识将有助于合理设计和选择胰岛素产生细胞。

Bioartificial pancreata promise physiological control of blood glucose levels, effectively treating Type I diabetes mellitus. Given the short supply of islets, development of alternative cell-lines appropriate for use in such devices is necessary. Transformed cells are a viable option, though their bioenergetic machinery may be inefficient. Because bioenergetics plays a role in the secretion of insulin, determining bioenergetic factors critical to insulin secretion may be important in selecting appropriate lines for inclusion into a bioartificial pancreas. This proposal will study cultures of entrapped transformed insulinoma cells and determine the impact of ATP and anaplerotic pathways on their ability to secrete insulin in response to an external glucose stimulus. NMR spectroscopic analysis of 13C and 31P data will provide a non-invasive quantitative assessment of the coupling between energy metabolism and insulin secretion, while non-NMR assays will derive glucose and oxygen consumption rates, as well as lactate, NH4+ and insulin production rates to provide a metabolic profile. Administration of specific inhibitors of the TCA cycle, anaplerotic pathways and oxidative phosphorylation, will aid in identifying factors critical to the initiation of insulin secretion. This knowledge will aid in the rational design and selection of insulin-producing cells.