Kathrin Heiss1, Luca Vanella2, Paolo Murabito3, Orazio Prezzavento2, Agostino Marrazzo2, Carlo Castruccio Castracani1, Ignazio Barbagallo2, Agata Zappalà1, Emanuela Arena2, Marinella Astuto3, Antonino Giarratano4, Giovanni Li Volti5. 1. Department of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95100 Catania, Italy. 2. Department of Drug Sciences, University of Catania, V.le A. Doria 6, 95125 Catania, Italy. 3. Azienda Ospedaliera Universitaria Policlinico "G. Rodolico", University of Catania, Via S. Sofia 78, 95125 Catania, Italy. 4. Department of Biopathology and Medical Biotechnologies (DIBIMED), Section of Anaesthesia, Analgesia, Intensive Care and Emergency, Paolo Giaccone University Hospital, University of Palermo, Via Del Vespro 38, 90100, Palermo, Italy. 5. Department of Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95100 Catania, Italy; Euro-Mediterranean Institute of Science and Technology, Via Michele Miraglia, 20, 90100 Palermo, Italy. Electronic address: livolti@unict.it.
Abstract
BACKGROUND: Sigma-1 receptors (σ1R) are highly expressed in neurons as well as microglia and have been shown to modulate the inflammatory response in the central nervous system and thus may serve as possible target for neuroprotective strategies. The aim of the present study was to test the effect of (+)-pentazocine, a putative σ 1R agonist, in an in vitro model of microglia activation. METHODS: Microglia (BV2 cells) was exposed (3h) to 1% oxygen and reoxygenation was allowed for 24h. Cells were treated with different concentrations (1, 10, 25 and 50μM) of (+)-pentazocine in the presence or absence of NE-100 (1μM), a well established σ1R antagonist. Cell viability and apoptosis were measured by cytofluorimetric analysis, whereas oxidative stress was evaluated by reduced glutathione (GSH) content and mitochondrial potential analysis. RESULTS: Our results showed that (+)-pentazocine was able to increase cell viability and restore mitochondrial potential at all concentrations whereas only 1 and 10μM were able to reduce significantly apoptotic cell death, to restore reduced glutathione intracellular content and prevent ERK1/2 phosphorylation. All these effects were abolished by concomitant treatment with NE-100. CONCLUSIONS: (+)-pentazocine exhibits significant dose dependent protective effects in our in vitro model of microglial activation thus suggesting that σ1R may represent a possible target for neuroprotection.
BACKGROUND:Sigma-1 receptors (σ1R) are highly expressed in neurons as well as microglia and have been shown to modulate the inflammatory response in the central nervous system and thus may serve as possible target for neuroprotective strategies. The aim of the present study was to test the effect of (+)-pentazocine, a putative σ 1R agonist, in an in vitro model of microglia activation. METHODS: Microglia (BV2 cells) was exposed (3h) to 1% oxygen and reoxygenation was allowed for 24h. Cells were treated with different concentrations (1, 10, 25 and 50μM) of (+)-pentazocine in the presence or absence of NE-100 (1μM), a well established σ1R antagonist. Cell viability and apoptosis were measured by cytofluorimetric analysis, whereas oxidative stress was evaluated by reduced glutathione (GSH) content and mitochondrial potential analysis. RESULTS: Our results showed that (+)-pentazocine was able to increase cell viability and restore mitochondrial potential at all concentrations whereas only 1 and 10μM were able to reduce significantly apoptotic cell death, to restore reduced glutathione intracellular content and prevent ERK1/2 phosphorylation. All these effects were abolished by concomitant treatment with NE-100. CONCLUSIONS:(+)-pentazocine exhibits significant dose dependent protective effects in our in vitro model of microglial activation thus suggesting that σ1R may represent a possible target for neuroprotection.