Introduction: Serotonin (5-HT) is involved in different pathways of the immune regulation. A dysmetabolism of the 5-HT system may be related to different pathologies, such as infectious and autoimmune diseases. In this preliminary study horse mixed-leukocytes were exposed to H2O2, responsible of a moderate mortality and growth inhibition, in order to evaluate the possible influence of experimental oxidative stress on 5-HT transporter (SERT), 5-HT2A and 5-HT1B receptors. Methodology: Mixed-leukocytes were obtained from blood samples of healthy horses by separation with ficoll. Total RNA was extracted through Trizol kit and 1 microgram was retro transcribed into cDNA for RTq PCR evaluation of SERT, 5-HT2A and 5-HT1B. Beta-actin was used as endogenous control. Results were expressed according to ΔΔCt calculation and by a relative quantization software. Results: RTq PCR results showed a SERT mRNA increase after 3 h exposition to H2O2 (2 micromol) and a decrease after 18 h or to H2O2 (10 micromol) vs control. Both the expressions of 5-HT2A and 5-HT1B receptors increased after 3 h exposition to H2O2 (2 micromol) and fell down under control values after 18 h or to H2O2 (10 micromol). Conclusions: Exposition to a lower concentration of H2O2 may be responsible for cell apoptosis. It is well known that H2O2 generation requires 5-HT internalization through an active transport mechanism into the cell and its degradation by monoamine oxidase A. Exposition to H2O2 at higher concentrations or for a longer time could have produced inhibition of transport mechanisms and peroxidation of membrane lipids leading to a lowered receptor density. These results pave the way for further investigations on the effect of both physiological and pathological stress in leukocytes.

Effect of oxidative stress on the expression of serotonin transporter, 5-HT2A and 5-HT1B receptors in horse leukocytes

BRUSCHETTA, Giuseppe;ZANGHI', GABRIELLA;D'ASCOLA, ANGELA;FERLAZZO, Alida
2015

Abstract

Introduction: Serotonin (5-HT) is involved in different pathways of the immune regulation. A dysmetabolism of the 5-HT system may be related to different pathologies, such as infectious and autoimmune diseases. In this preliminary study horse mixed-leukocytes were exposed to H2O2, responsible of a moderate mortality and growth inhibition, in order to evaluate the possible influence of experimental oxidative stress on 5-HT transporter (SERT), 5-HT2A and 5-HT1B receptors. Methodology: Mixed-leukocytes were obtained from blood samples of healthy horses by separation with ficoll. Total RNA was extracted through Trizol kit and 1 microgram was retro transcribed into cDNA for RTq PCR evaluation of SERT, 5-HT2A and 5-HT1B. Beta-actin was used as endogenous control. Results were expressed according to ΔΔCt calculation and by a relative quantization software. Results: RTq PCR results showed a SERT mRNA increase after 3 h exposition to H2O2 (2 micromol) and a decrease after 18 h or to H2O2 (10 micromol) vs control. Both the expressions of 5-HT2A and 5-HT1B receptors increased after 3 h exposition to H2O2 (2 micromol) and fell down under control values after 18 h or to H2O2 (10 micromol). Conclusions: Exposition to a lower concentration of H2O2 may be responsible for cell apoptosis. It is well known that H2O2 generation requires 5-HT internalization through an active transport mechanism into the cell and its degradation by monoamine oxidase A. Exposition to H2O2 at higher concentrations or for a longer time could have produced inhibition of transport mechanisms and peroxidation of membrane lipids leading to a lowered receptor density. These results pave the way for further investigations on the effect of both physiological and pathological stress in leukocytes.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11570/3106515
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