caa a22 4500 475772083 CHVBK 20180406123620.0 cr unu---uuuuu 170329e20001001xx s 000 0 eng 10.1007/s001250051520 doi (NATIONALLICENCE)springer-10.1007/s001250051520 High glucose modulates P2X7 receptor-mediated function in human primary fibroblasts [Elektronische Daten] [A. Solini, P. Chiozzi, S. Falzoni, A. Morelli, R. Fellin, F. Di Virgilio] Aims/hypothesis. Purinergic receptors are a family of newly characterized plasma membrane molecules involved in several and as yet only partially known cellular functions such as vascular reactivity, apoptosis and cytokine secretion. Little is known about the effect extracellular microenvironment has on their function. Fibroblasts share several features with smooth muscle cells and are an important constituent of the atherosclerotic plaque. Our aim was to evaluate the effect of high glucose concentration on ATP-mediated responses in human fibroblasts.¶Methods. Fibroblasts were obtained by skin biopsies and grown at two different glucose concentrations. We evaluated receptor expression by RT-PCR and immunoblotting and receptor localization by immunofluorescence. Plasma membrane potential and calcium changes were measured by fluorescent indicators. Apoptosis was determined by ethidium bromide staining and caspase-3 activation.¶Results. We show that cells grown in a medium with high glucose concentration underwent great ATP-mediated morphological changes, enhanced apoptosis, caspase 3 activation and interleukin-6 release. We identified P2X7 as the main purinergic receptor involved in these responses. Furthermore, high glucose concentration triggered the assembly of P2X7 into ring-like structures located at the periphery of the cells.¶Conclusion/interpretation. Given that ATP is frequently released into the extracellular milieu upon cell and tissue damage, secretory exocytosis or activation of plasma membrane transporters, we hypothesize that ATP receptors participate in the pathogenesis of vascular complications of diabetes. [Diabetologia (2000) 43: 1248-1256] Springer-Verlag Berlin Heidelberg, 2000 Keywords Calcium, interleukin-6, P2 purinergic receptor, apoptosis, hyperglycaemia nationallicence Solini A. Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Ferrara, Ferrara, Italy, IT aut Chiozzi P. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut Falzoni S. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut Morelli A. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut Fellin R. Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Ferrara, Ferrara, Italy, IT aut Di Virgilio F. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut https://doi.org/10.1007/s001250051520 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s001250051520 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Solini A. Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Ferrara, Ferrara, Italy, IT aut NATIONALLICENCE 700 1- Chiozzi P. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut NATIONALLICENCE 700 1- Falzoni S. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut NATIONALLICENCE 700 1- Morelli A. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut NATIONALLICENCE 700 1- Fellin R. Department of Clinical and Experimental Medicine, Section of Internal Medicine, University of Ferrara, Ferrara, Italy, IT aut NATIONALLICENCE 700 1- Di Virgilio F. Department of Experimental and Diagnostic Medicine, Section of General Pathology, University of Ferrara, Ferrara, Italy, IT aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer