Implication of reactive oxygen species, nitric oxide, and their imbalance in the pathogenesis of chronic kidney disease

活性氧、一氧化氮及其失衡在慢性肾脏病发病机制中的意义

基本信息

批准号：
17590852
负责人：
KASHIHARA Naoki
金额：
$ 2.24万
依托单位：
Kawasaki Medical School
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

项目摘要

Endothelial dysfunction is a well-established pathogenetic mechanism underlying the development of vascular and renal complications in the various forms of renal injuries and aging kidney as well. The aim of this study is to elucidate the pathogenetic mechanism of renal injury and explore the possible mechanism that could underlie endothelial dysfunction in the systemic vasculature in the progressive forms of renal injuries and senescent kidney. We hypothesized that an imbalance between reactive oxygen species (ROS) and nitric oxide (NO) in the vasculature is implicated in both renal injury and macrovascular dysfunction.We have successfully developed the novel method by which one could observe the generation of ROS and NO in situ. Briefly, under general anesthesia, The rat whole body was perfused by the infusion pump with 37℃ phosphate buffer saline (PBS) at a flow rate of 5ml/min. Once blood had been removed, the whole body was perfused with PBS containing 0.01mmol/L diaminorhodamine- … More 4M AM (DAR-4M AM), 0.05mmol/L dichlorodihydrofluorescin diaceate (DCFH-DA), 0.1mmol/L L-Arginine, and 2mmol/L CaCl2 for 10 minutes at a flow rate of 3ml/min. To remove the unreacted reagent and fix tissues, a postperfusion was added with 4% paraformaldehyde containing 0.2mmol/L N□-nitro-L-arginine methyl ester hydrochloride (L-NAME ; Sigma-Aldrich Japan) for 30 minutes at a flow rate of 5ml/min. Fluorescent images of ROS and NO were obtained with a confocal laser-scanning microscopy TCS-NT (Leica-Microsystems, Tokyo, Japan). The wavelength was as follow ; DAR-4M AM, excitation at 560nm and emission at 575nm ; DCFH-DA, excitation at 490nm and emission at 530nm.We have discovered increased generation of ROS and deceased baioavailable NO in the renal tissue in the 5/6 nephrectomized rats. Accumulation of nitrotyrosine, a trace of nitrosative stress, was also increased in the renal vasculature and aorta We also found NADPH oxidase activity was implicated in the generation of ROS in this model. Angiotensin receptor blocker inhibited increased productions of ROS and NO, and significantly reduced nitrosative stress Less

内皮功能障碍是各种形式的肾损伤和衰老肾脏中血管和肾脏并发症发生的一个明确的致病机制，本研究的目的是阐明肾损伤的致病机制并探讨其可能的机制。在进行性肾损伤和衰老肾脏中，可能是全身脉管系统内皮功能障碍的基础。我们发现脉管系统中活性氧（ROS）和一氧化氮（NO）之间的不平衡。我们成功地开发了一种可原位观察ROS和NO生成的新方法，简而言之，在全身麻醉下，用输液泵对大鼠全身进行37℃磷酸盐灌注。以 5ml/min 的流速去除血液后，用含有 0.01mmol/L 二氨基罗丹明的 PBS 灌注全身。 4M AM。 (DAR-4M AM)、0.05mmol/L二乙酸二氯二氢荧光素(DCFH-DA)、0.1mmol/L L-精氨酸和2mmol/L CaCl2，以3ml/min的流速作用10分钟，除去未反应的试剂。固定组织，灌注后加入含0.2mmol/L的4%多聚甲醛N□-硝基-L-精氨酸甲酯盐酸盐（L-NAME；Sigma-Aldrich Japan）以5ml/min的流速使用共聚焦激光扫描显微镜TCS-获得ROS和NO的荧光图像。 NT (Leica-Microsystems, Tokyo, Japan) 波长如下：DAR-4M AM，激发波长为 560nm，发射波长为575nm；DCFH-DA，在490nm处激发，在530nm处发射。我们发现5/6肾切除大鼠的肾组织中ROS的产生增加，并且硝基酪氨酸（微量亚硝化应激）的积累也增加。在脉管系统和主动脉肾中我们还发现 NADPH 氧化酶活性与在该模型中，血管紧张素受体阻滞剂抑制了 ROS 和 NO 的产生，并显着减少了亚硝化应激。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Amelioration of progressive renal injury by genetic manipulation of Klotho gene

DOI：
10.1073/pnas.0611079104
发表时间：
2007-02-13
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
影响因子：
11.1
作者：
Haruna, Yoshisuke;Kashihara, Naoki;Kanwar, Yashpal S.
通讯作者：
Kanwar, Yashpal S.

Isohumulones derived from hops ameliorate renal injury via an anti-oxidative effect in Dahl salt-sensitive rats