GLUTATHIONE AND MITOCHONDRIA IN TOXIC RENAL INJURY

谷胱甘肽和线粒体功能在 保护细胞免受损伤和维持细胞 动态平衡。肾脏容易受到许多有毒物质的伤害。 化学物质和病理状态。肾单位的所有节段， 然而，它们对伤害并不同样敏感。校长 这项研究的目的是刻画性格特征和 谷胱甘肽在肾脏线粒体中的作用及其机制研究 肾组织谷胱甘肽状态及线粒体功能的影响 受伤。分离的线粒体和纯化或高度浓缩的细胞 男性Fischer肾单位单个节段的悬吊 将使用344只大鼠。因为他们的优势， 因此，为了便于隔离，将进行初步研究 有近端肾小管细胞和肾皮质线粒体。这个 具体问题有：1)肾的起源是什么 线粒体谷胱甘肽及其处置是如何调节的？ 2)谷胱甘肽在肾近端小管中的作用 线粒体功能？3)谷胱甘肽的状态和 线粒体功能在肾近端易感性中的作用 肾小管上皮细胞与化学损伤的关系？4)可以开发程序吗？ 为了分离高度丰富的粗大上升肢体种群， 远端肾小管和皮质收集肾小管细胞，以及如何 比较了它们的生化特性和对伤害的敏感性 近端肾小管上皮细胞的？Percoll密度梯度 离心法将用于细胞纯化。数字化 分离的细胞将被用来分离胞浆 和线粒体，使更直接地研究线粒体成为可能 功能。选择性改变谷胱甘肽浓度的药物 和氧化还原状态以及改变线粒体生物能量学的那些 将被用来探索谷胱甘肽在 线粒体功能。使用会引起化学性损伤 各种有毒化学物质，以及谷胱甘肽和 线粒体氧化活性在防护和敏感性中的作用 对损伤会产生过氧化氢、甲基乙烯基酮、S-(2- 氯乙基)-DL-半胱氨酸)和需要代谢的化合物 激活(止痛剂扑热息痛、抗生素 头孢菌素--细胞损伤易感性的碱基状态 也将接受调查。这项研究将有助于提供更好的 对肾脏药物代谢和生化的认识 化学物质产生肾毒性的机制。

Glutathione and mitochondrial functions play important roles in protecting cells from injury and in maintenance of cellular homeostasis. The kidneys are susceptible to injury from many toxic chemicals and pathological states. All segments of the nephron, however, are not equally sensitive to injury. The principal objective of this research is to characterize the disposition and role of glutathione in renal mitochondrial and to study the influence of glutathione status and mitochondrial function in renal injury. Isolated mitochondria and purified or highly enriched cell suspensions of individual segments of the nephron from male Fischer 344 rats will be used. Because of their predominance and, therefore, ease of isolation, initial studies will be conducted with proximal tubular cells and renal cortical mitochondria. The specific questions addressed are: 1) What is the origin of renal mitochondrial glutathione and how is its disposition regulated? 2) What is the role of glutathione in renal proximal tubular mitochondrial function? 3) What role do glutathione status and mitochondrial function play in the susceptibility of renal proximal tubular cells to chemical injury? 4) Can procedures be developed to isolate highly enriched populations of thick ascending limb, distal tubular, and cortical collecting tubular cells, and how do their biochemical properties and susceptibility to injury compare to those of proximal tubular cells? Percoll density-gradient centrifugation will be used in cell purification. Digitonin fractionation of isolated cells will be used to separate cytosol and mitochondria, enabling more direct study of mitochondrial function. Agents that selectively alter glutathione concentrations and redox status and those that alter mitochondrial bioenergetics will be employed to explore the role of glutathione in mitochondrial function. Chemical injury will be induced by the use of various toxic chemicals, and the role of glutathione and mitochondrial oxidative activity in protection from and sensitivity to injury will be hydroperoxide, methyl vinyl ketone, S-(2- chloroethyl)-DL-cysteine) and compounds requiring metabolic activation (the analgesic acetaminophen, the antibiotic cephaloridine, the base status on cellular susceptibility to injury will also be investigated. The research will help provide a better understanding of renal drug metabolism and the biochemical mechanisms by which chemicals produce nephrotoxicity.