UROLITHIASIS--ROLE OF NEPHRON DYSFUNCTION/INJURY

尿石症——肾单位功能障碍/损伤的作用

• 批准号: 6634973
• 负责人: JULIE A. JONASSEN
• 金额: $ 34.85万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-01-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6634973
• 关键词: acid base balance; biological signal transduction; calcium flux; catalase; cell line; cell membrane; cell proliferation; cellular pathology; cytotoxicity; enzyme activity; free radical oxygen; gene expression; genetic manipulation; genetic transcription; glutathione peroxidase; kidney function; laboratory rat; messenger RNA; mitochondria; mitogen activated protein kinase; northern blottings; oxalates; pathologic process; protein kinase; regulatory gene; renal tubule; superoxide dismutase; tissue /cell culture; transcription factor; transfection; urinary calculi

Our working hypothesis is that oxalate-induced injury to renal epithelial cells plays a central role in the development of calcium oxalate stone disease, with oxalate-induced injury promoting the nucleation, growth and retention of calcium oxalate crystals within the kidney. Recent studies in our laboratory support this hypothesis and indicate that oxalate-induced injury to renal tubular cells may be secondary to increased free radical production. This working hypothesis serves as the focus of the present proposal and its 3 interrelated aims. Aim 1 will determine the mechanism(s) of oxalate toxicity, defining a) the role of oxalate crystals and/or the oxalate anion in oxalate injury, b) cellular processes that could mediate oxalate actions c) relevant sites of free radical production within the cells and d) targets of free radical actions (plasma membrane, mitochondria, etc.) e) the effects of prior "sensitizing" stimuli on oxalate toxicity. Aim 2 will characterize oxalate-induced changes in gene expression in renal epithelial cells, defining a) the pattern of oxalate induced changes in gene expression, b) the cellular events leading to the observed changes in gene expression, and c) the therapeutic benefits of genetic manipulations which increase free radical "buffering" or facilitate oxalate degradation. Aim 3 will determine whether hyperoxaluria in experimental animals will evoke changes in renal cell function which are comparable to those observed in response to hyperoxaluria in cultured renal epithelial cells. The proposed in vitro studies will utilize LLC-PK1 cells for the majority of the studies confirming key findings in distal tubular cells (MDCK cells) and in human renal epithelial cells (RPTEC cells). These studies will define oxalate actions in vitro and evaluate the effectiveness of various pharmacological and genetic manipulations to limit oxalate toxicity. Results from these in vitro studies will inform and guide the design of in vivo studies to assess the consequences of hyperoxaluria in experimental animals and to define the role of oxidant stress in the development/progression of stone disease. This combination of in vitro and in vivo studies should provide important insights as to processes involved in calcium oxalate stone disease and suggest possible therapeutic strategies for management of this disease.

我们的工作假设是，草酸盐诱导的肾上皮细胞损伤在草酸钙结石病的发展中起着核心作用，草酸盐诱导的损伤促进草酸钙晶体在肾脏内的成核、生长和保留。 我们实验室最近的研究支持了这一假设，并表明尿酸盐诱导的肾小管细胞损伤可能是继发于自由基产生增加。 这一工作假设是本提案及其三个相互关联的目标的重点。 目的1将确定草酸盐毒性的机制，定义a）草酸盐晶体和/或草酸盐阴离子在草酸盐损伤中的作用，B）可能介导草酸盐作用的细胞过程，c）细胞内自由基产生的相关位点，以及d）自由基作用的靶点（质膜、线粒体等）。e）先前的"致敏"刺激对草酸盐毒性的影响。 目的2将描述草酸盐诱导的肾上皮细胞基因表达的变化，定义a）草酸盐诱导的基因表达变化的模式，B）导致观察到的基因表达变化的细胞事件，和c）增加自由基"缓冲"或促进草酸盐降解的遗传操作的治疗益处。 目的3将确定实验动物的高尿酸是否会引起肾细胞功能的变化，这些变化与在培养的肾上皮细胞中观察到的高尿酸反应相当。 拟定的体外研究将使用LLC-PK1细胞进行大部分研究，以确认远端肾小管细胞（MDCK细胞）和人肾上皮细胞（RPTEC细胞）中的关键发现。 这些研究将定义草酸盐的体外作用，并评估各种药理学和遗传操作限制草酸盐毒性的有效性。 这些体外研究的结果将为体内研究的设计提供信息和指导，以评估实验动物高尿酸的后果，并确定氧化应激在结石病发展/进展中的作用。 这种体外和体内研究的结合应提供重要的见解，草酸钙结石疾病的过程中所涉及的，并建议可能的治疗策略，这种疾病的管理。

项目成果

Oxalate transport in a line of porcine renal epithelial cells--LLC-PK1 cells.

猪肾上皮细胞系——LLC-PK1细胞中的草酸盐转运。

• DOI: 10.1016/s0022-5347(17)32869-0
• 发表时间: 1994
• 期刊: The Journal of urology
• 作者: Ebisuno,S;Koul,H;Menon,M;Scheid,C
• 通讯作者: Scheid,C

How elevated oxalate can promote kidney stone disease: changes at the surface and in the cytosol of renal cells that promote crystal adherence and growth.

• DOI: 10.2741/1265
• 发表时间: 2004
• 期刊: Frontiers in bioscience : a journal and virtual library
• 作者: C. Scheid;Lu-Cheng Cao;T. Honeyman;J. Jonassen

Oxalate-induced damage to renal tubular cells.

草酸诱导的肾小管细胞损伤。

• 发表时间: 1995
• 期刊: Scanning microscopy.
• 作者: Scheid,CR;Koul,HK;Kennington,L;Hill,WA;Luber-Narod,J;Jonassen,J;Honeyman,T;Menon,M
• 通讯作者: Menon,M

Targeted overexpression of insulin-like growth factor I to osteoblasts of transgenic mice: increased trabecular bone volume without increased osteoblast proliferation.

• DOI: 10.1210/endo.141.7.7585
• 发表时间: 2000-07
• 期刊: Endocrinology
• 影响因子: 4.8
• 作者: Guisheng Zhao;M. Monier‐Faugere;M. Langub;Z. Geng;T. Nakayama;J. Pike;S. Chernausek;C. Rosen;L. Donahue;H. Malluche;J. Fagin;T. Clemens;T. Clemens

Oxalate toxicity in LLC-PK1 cells, a line of renal epithelial cells

• DOI: 10.1016/s0022-5347(01)66402-4
• 发表时间: 1996-03-01
• 期刊: JOURNAL OF UROLOGY
• 影响因子: 6.6
• 作者: Scheid, C;Koul, H;Menon, M
• 通讯作者: Menon, M