Participation of NADPH Oxidase 5 in hypoxia-induced generation of reactive oxygen species in human endothelial cells.

ALVAREZ MS; MARIELA NOLLY.; MAZZEI L; SALVARREDI L; QUESADA I; CASTRO C

Congreso; XL Reunión Científica Anual de la Sociedad de Biología de Cuyo.; 2022

021- PARTICIPATION OF NADPH OXIDASE 5 IN HYPOXIA-INDUCED GENERATION OFREACTIVE OXYGEN SPECIES IN HUMAN ENDOTHELIAL CELLSÁlvarez, MS1; Nolly M1, Mazzei L1; Salvarredi L 1-2, Quesada I1 and Castro C11Instituto de Bioquímica y Biotecnología, FCM UNCUYO e IMBECU-CONICET, and 2Fundación Escuela de Medicina Nuclear;Comisión Nacional de Energía Atómica- Instituto Balseiro, UNCuyoe-mail: msoledadalvarez@gmail.comNADPH oxidase (NOX) is the main producer of reactive oxygen species (ROS) that may contribute to the pathogenesis of endothelialdysfunction (ED). In endothelial cells, ROS can be generated from sources such as NOX and mitochondria, which in turn can serve assignaling molecules in a wide variety of processes including posttranslational modification of proteins involved in Ca2+ homeostasis. Therole of NOX5. a calcium-activated NOX isoform, in triggering ROS in ED is not clear. The aim of this work was to determine theimplication of NOX5 in hypoxia aside from pro-atherogenic/pro-inflammatory conditions. Human umbilical vein endothelial cells(HUVECs) were treated with Angiotensin II (AngII; 10-7 M), TNF-alpha (100 ng/mL) or cobalt chloride (CoCl2), a hypoxia mimeticagent. NOX5 expression was first analyzed by RT-PCR and western blot. Then, Superoxide generation was measured by fluorescencetechniques using 5 uM dihydroethidium (DHE) for 30 min in the dark at 37 °C. An increase in NOX5 protein expression was found inHUVECs stimulated with pro-inflammatory factors, and both significantly increase the expression of monocyte chemoattractant protein 1(MCP-1) and Interleukin 32 (IL-32) (**p