Peroxynitrite, protein nitration and advanced diabetic neuropathy

过氧亚硝酸盐、蛋白质硝化和晚期糖尿病神经病变

• 批准号: 8625363
• 负责人: Mark A. Yorek
• 金额: $ 17.28万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8625363
• 关键词: Peroxynitrite; protein; nitration; advanced; diabetic

DESCRIPTION (provided by applicant): Evidence for important role of peroxynitrite, a product of superoxide reaction with nitric oxide, in diabetic complications is emerging. Using new pharmacological agents i.e., peroxynitrite decomposition catalysts and protein nitration inhibitor, and iNOS-knockout mice, we obtained findings implicating peroxynitrite injury in toto and protein nitration in early experimental peripheral diabetic neuropathy (PDN). We found that 1) nitro- tyrosine (NT), a footprint of peroxynitrite injury, accumulates in peripheral nerve, spinal cord, and DRG neurons of streptozotocin (STZ)-diabetic and ob/ob mice; 2) STZ-diabetic iNOS-deficient mice do not develop nerve conduction deficits and have less severe sensory neuropathy compared with diabetic wild-type mice; and 3) peroxynitrite decomposition catalysts and, to a lesser extent, a protein nitration inhibitor, corrected nerve conduction deficits and sensory neuropathy in mice with Type 1 and Type 2 diabetes. We also found that Type 2 diabetic patients accumulate different amounts of nitrated proteins in peripheral blood monocytes, that monocyte NT concentration is ~ 75% greater in Type 2 diabetic subjects compared with non-diabetic group, and that skin production of NO, a precursor of peroxynitrite, is reduced by pioglitazone treatment. Others showed that increased plasma NT content correlated with endothelial dysfunction and redistribution of sudomotor responses, an early sign of sympathetic nerve dysfunction, in diabetic patients. The overall objective of this proposal is to dissect the roles of peroxynitrite and protein nitration in functional and morphological changes of advanced experimental PDN, and to determine if NT levels in serum, peripheral blood monocytes, and skin can be used as biomarkers of the presence, severity, and progression of PDN in human subjects with diabetes. The specific aims are: 1) evaluate if peroxynitrite decomposition catalyst and protein nitration inhibitor reverse functional, behavioral, and morphological changes of advanced PDN in STZ- diabetic and Akita mice; 2) assess the effect of blood glucose control on accumulation and disappearance of NT in tissue-sites of PDN, skin, and circulation; and 3) determine potential values of serum, peripheral blood monocyte, and skin NT levels as biomarkers of the presence, severity, and progression of PDN in human subjects with diabetes mellitus. The findings will advance our understanding of the pathogenesis of PDN, and may provide rationale for development of new therapeutics. They may also lead to identification of a new biomarker(s) of PDN with diagnostic and prognostic value. PUBLIC HEALTH RELEVANCE: Peripheral diabetic neuropathy (PDN) is the most devastating complication of diabetes mellitus, and a leading cause of foot amputation. Evidence for important role of peroxynitrite (a product of superoxide reaction with nitric oxide) in PDN is emerging and is supported by our preliminary data obtained in both diabetic mouse models and human subjects with diabetes mellitus. The overall objective of this proposal is to dissect the roles of peroxynitrite and one of its components, protein nitration, in functional and morphological changes of advanced experimental PDN, and to determine if nitrotyrosine levels in serum, peripheral blood monocytes, and skin can be used as biomarkers of the presence, severity, development and rate of progression of PDN in human subjects with diabetes. The specific aims are: 1) evaluate if peroxynitrite decomposition catalyst or protein nitration inhibitor reverse functional, behavioral, and morphological manifestations of advanced PDN in two mouse models of Type 1 diabetes; 2) assess the effect of blood glucose control on accumulation and disappearance of nitrotyrosine in tissue-sites of PDN, skin, and circulation; and 3) determine potential values of serum, peripheral blood monocyte, and skin nitrotyrosine levels as biomarkers of the presence, severity, development, and rate of progression of PDN in human subjects with diabetes mellitus. The findings will advance our understanding of the pathogenesis of PDN, and may provide rationale for development of new therapeutics. They may also lead to identification of a new biomarker(s) of PDN with diagnostic and prognostic value.

描述（由申请人提供）：过氧亚硝酸盐的重要作用（超氧化物与一氧化氮的产物）在糖尿病并发症中的重要作用。使用新的药理学剂，即过氧亚硝酸盐分解催化剂和蛋白硝化抑制剂，以及iNOS-KNOCKOUT小鼠，我们获得了在早期实验性外周糖尿病神经疗法（PDN）中涉及过氧亚硝酸盐损伤的发现。我们发现1）硝基酪氨酸（NT），过氧亚硝酸盐损伤的足迹，在周围神经，脊髓和DRG神经元中积聚，链霉菌素（STZ） - 糖尿病和ob/ob/ob/ob/ob/ob/ob/ob/ob/ob/ob小鼠； 2）与糖尿病性野生型小鼠相比，STZ-糖尿病iNOS缺陷小鼠不会出现神经传导缺陷，并且具有严重的感觉神经病； 3）过氧亚硝酸盐分解催化剂，并且在较小程度上是蛋白硝化抑制剂，在患有1型和2型糖尿病的小鼠中校正了神经传导缺陷和感觉神经病。我们还发现，2型糖尿病患者在外周血单核细胞中积累了不同量的硝酸蛋白，与非糖尿病组相比，单核细胞NT浓度在2型糖尿病患者中高约75％，并且通过Pioglitaz酮治疗减少了皮肤的NO，PROCSYNITRITE的皮肤产生。其他人则表明，糖尿病患者的血浆NT含量与内皮功能障碍和Sudomotor反应的重新分布相关的血浆NT含量增加和Sudomotor反应的重新分布。该提案的总体目的是剖析过氧亚硝酸盐和蛋白硝化在晚期实验PDN功能和形态变化中的作用，并确定是否可以将NT水平用于血清，外周血单核细胞和皮肤是否可以用作人类受试者PDN的存在，严重性和PDN的生物标志物。具体目的是：1）评估过氧亚硝酸盐分解催化剂和蛋白硝化抑制剂反向功能，行为和形态学变化在STZ-糖尿病和秋田小鼠中的形态变化； 2）评估血糖控制对PDN，皮肤和循环组织中NT的积累和消失的影响； 3）确定血清，外周血单核细胞和皮肤NT水平的潜在值，作为糖尿病患者PDN的存在，严重程度和进展的生物标志物。这些发现将提高我们对PDN发病机理的理解，并可能为开发新疗法提供理由。它们还可能导致鉴定具有诊断和预后价值的新生物标志物。 公共卫生相关性：周围糖尿病神经病（PDN）是糖尿病最具破坏性的并发症，是脚足截肢的主要原因。 PDN中过氧亚硝酸盐（超氧化物反应与一氧化氮的产物）重要作用的证据正在出现，并且得到了我们在糖尿病小鼠模型和糖尿病的人类受试者中获得的初步数据的支持。 The overall objective of this proposal is to dissect the roles of peroxynitrite and one of its components, protein nitration, in functional and morphological changes of advanced experimental PDN, and to determine if nitrotyrosine levels in serum, peripheral blood monocytes, and skin can be used as biomarkers of the presence, severity, development and rate of progression of PDN in human subjects with diabetes.具体目的是：1）评估过氧亚硝酸盐分解催化剂或蛋白硝化抑制剂反向功能，行为和形态学表现是否在两种型型糖尿病的小鼠模型中； 2）评估血糖控制对PDN，皮肤和循环组织中硝基酪氨酸的积累和消失的影响； 3）确定血清，外周血单核细胞和皮肤亚硝基酪氨酸水平的潜在值，作为在糖尿病患有糖尿病的人类受试者中存在，严重程度，发育和PDN进展速率的生物标志物。这些发现将提高我们对PDN发病机理的理解，并可能为开发新疗法提供理由。它们还可能导致鉴定具有诊断和预后价值的新生物标志物。