Identification of the physiological function of the NADPH oxidase Nox4 in the kidney

肾脏NADPH氧化酶Nox4的生理功能鉴定

• 批准号: 531817008
• 负责人: Dr. Flávia Rezende
• 金额: --
• 依托单位: Institut für Kardiovaskuläre Physiologie (Physiologie I)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/531817008?language=en
• 关键词: Identification; physiological; function; NADPH; oxidase

The only known enzymatic function of NADPH oxidase enzymes of the Nox-family is the production of reactive oxygen species (ROS: H2O2 or O2•-). Nox enzymes differ in their regulation, site of expression and type of ROS produced. The isoform Nox4, differently to all other Nox isoforms, is constitutively active and under physiological conditions almost exclusively expressed in the kidney. Despite intense research, the renal function of Nox4 is, however, completely unknown. The aim of the present application is to uncover this function focusing on transport processes in the proximal tubule of the kidney. Preliminary untargeted metabolomics from plasma, urine and renal tissue of wild type and Nox4-/- mice suggested that the enzyme controls metabolism of sulphur amino acids and/or reabsorption of vitamins of the complex B. In this application, these aspects will be further explored, largely in vivo, using global and kidney-specific inducible Nox4 knockout mice. Metabolites will be analysed by targeted LC-MS/MS from plasma and urine of mice with low sulphur and vitamin B-deficient diet interventions. Moreover, the intracellular localization and interaction partners of the enzyme will be identified using a Nox4-FLAG knock-in mouse, generated in preparation for this application. In summary, the project will identify the physiological function of Nox4 in the kidney using in vivo models.

Nox家族的NADPH氧化酶的唯一已知酶功能是产生活性氧（ROS：H2 O2或O2·-）。Nox酶在其调节、表达位点和产生的ROS类型方面不同。与所有其他Nox同种型不同，同种型Nox 4是组成型活性的，并且在生理条件下几乎仅在肾脏中表达。尽管进行了大量的研究，但Nox 4的肾功能仍然完全未知。本申请的目的是揭示聚焦于肾近端小管中的转运过程的这种功能。来自野生型和Nox 4-/-小鼠的血浆、尿液和肾组织的初步非靶向代谢组学表明，该酶控制含硫氨基酸的代谢和/或复合物B的维生素的重吸收。在本申请中，这些方面将进一步探索，主要是在体内，使用全球和肾脏特异性诱导型Nox 4敲除小鼠。将通过靶向LC-MS/MS分析低硫和维生素B缺乏饮食干预小鼠的血浆和尿液中的蛋氨酸。此外，将使用Nox 4-FLAG敲入小鼠鉴定酶的细胞内定位和相互作用配偶体，所述小鼠是在为该应用做准备时产生的。总之，该项目将使用体内模型确定Nox 4在肾脏中的生理功能。