Kathrin Heiss1, Marco Raffaele2, Luca Vanella2, Paolo Murabito3, Orazio Prezzavento2, Agostino Marrazzo2, Giuseppina Aricò2, Carlo Castruccio Castracani1, Ignazio Barbagallo2, Agata Zappalà1, Roberto Avola1, Giovanni Li Volti4. 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 Biomedical and Biotechnological Sciences, University of Catania, Via S. Sofia 64, 95100 Catania, Italy; Euro-Mediterranean Institute of Science and Technology, Via Emerico Amari 131, 90100 Palermo, Italy. Electronic address: livolti@unict.it.
Abstract
BACKGROUND: Sigma receptors (σ1R) are expressed both in neurons and microglia and can be considered as a promising target for developing pharmacological strategies for neuroprotection in various experimental models. The aim of the present study was to test the effect of (+)-pentazocine, a putative σ 1R agonist, in an in vitro model of neuron/microglia crosstalk following hypoxia/reoxygenation. METHODS: Microglia (BV2 cells) was exposed (3h) to 1% oxygen and reoxygenation was allowed for 24h. Conditioned media obtained from this experimental condition was used to treat neuroblast-like cell line (SH-SY5Y cells) in the presence or absence of (+)-pentazocine (25μM). Cell viability was measured by cytofluorimetric analysis, whereas inflammation and oxidative stress were evaluated by the expression of Hsp70, GAD, SOD and p65. Microglial cell migration was also evaluated by Xcelligence technology. RESULTS: Our results showed that (+)-pentazocine was able to increase SH-SY5Y cell viability following exposure to microglial-conditioned medium. Furthermore, (+)-pentazocine was also able to inhibit microglial cell toward neuron treated with hypoxic conditioned medium. Finally, pharmacological treatment reduced the expression of inflammatory and oxidative stress markers (GAD, SOD and p65). Interestingly, hypoxic medium was able to reduce the expression of Hsp70 and such effect was prevented by (+)-pentazocine treatment. CONCLUSIONS: (+)-Pentazocine exhibits significant neuroprotective effects in our in vitro model of SH-SY5Y/microglial crosstalk thus suggesting that σ1R may represent a possible strategy for neuroprotection.
BACKGROUND: Sigma receptors (σ1R) are expressed both in neurons and microglia and can be considered as a promising target for developing pharmacological strategies for neuroprotection in various experimental models. The aim of the present study was to test the effect of (+)-pentazocine, a putative σ 1R agonist, in an in vitro model of neuron/microglia crosstalk following hypoxia/reoxygenation. METHODS: Microglia (BV2 cells) was exposed (3h) to 1% oxygen and reoxygenation was allowed for 24h. Conditioned media obtained from this experimental condition was used to treat neuroblast-like cell line (SH-SY5Y cells) in the presence or absence of (+)-pentazocine (25μM). Cell viability was measured by cytofluorimetric analysis, whereas inflammation and oxidative stress were evaluated by the expression of Hsp70, GAD, SOD and p65. Microglial cell migration was also evaluated by Xcelligence technology. RESULTS: Our results showed that (+)-pentazocine was able to increase SH-SY5Y cell viability following exposure to microglial-conditioned medium. Furthermore, (+)-pentazocine was also able to inhibit microglial cell toward neuron treated with hypoxic conditioned medium. Finally, pharmacological treatment reduced the expression of inflammatory and oxidative stress markers (GAD, SOD and p65). Interestingly, hypoxic medium was able to reduce the expression of Hsp70 and such effect was prevented by (+)-pentazocine treatment. CONCLUSIONS:(+)-Pentazocine exhibits significant neuroprotective effects in our in vitro model of SH-SY5Y/microglial crosstalk thus suggesting that σ1R may represent a possible strategy for neuroprotection.
Authors: J Wang; J Zhao; X Cui; B A Mysona; S Navneet; A Saul; M Ahuja; N Lambert; I G Gazaryan; B Thomas; K E Bollinger; S B Smith Journal: Free Radic Biol Med Date: 2019-02-10 Impact factor: 7.376