Glyoxylate, a new marker of type-2 diabetes, induces neutrophil activation

MARTIRE-GRECO D; RODRIGUEZ-RODRIGUES N; LANDONI V.I; CASTILLO LUIS; FERNÁNDEZ G C

Mar del Plata

Congreso; Congreso SAI/SAIC 2016; 2016

It has been shown that glyoxylate (Glx) levels become significantly elevated in a mouse model of diabetes and in type-2 diabetic patients, placing this new metabolic marker in the con text of diabetes pathology. As diabetes has been associated with inflammation inducing micro- and macro vascular damage, we hypothesize that Glx may be involved in diabetic-derived inflammation. Moreover, PMN activation has been associated with tissue injury in different pathophysiological conditions. Therefore, our aim was to investigate the direct role of Glx in neutrophil (PMN) responses. We evaluated activation of isolated human PMN by Glx (10 μg/ml) measuring the up-regulation of CD11b and ROS generation with Dihydrorhodamine by flow cytometry, NETs formation by confocal microscopy and chemokinesis using a Boyden chamber. We also evaluated the adhesion of Glx-treated PMN to endothelial cells (HMEC-1) by measuring the activity alkaline phosphatase (AP) with the substrate pnitrophenylphosphate (NPP) in the adhered PMN after extensive washing (Enzymatic Unit definition: 1 EU of AP=1 nmol of NPP/ min). We found an up-regulation of the adhesion marker CD11b after Glx treatment, associated with an increased chemokinesis and higher adhesion to endothelial cells (mean fluorescence intensity CD11b: Control= 293.7±9.0, Glx=469.1±76.0*; Nº of PMN migrated/field: Ctrol=10.1±1.6, Glx=22.4±2.2*; AP EU: Control= 32.4±3.0, Glx=51.4±3.3*, *p