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Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress

We obtained evidence that mouse BV2 microglia synthesize neurosteroids dynamically to modify neurosteroid levels in response to oxidative damage caused by rotenone. Here, we evaluated whether neurosteroids could be produced and altered in response to rotenone by the human microglial clone 3 (HMC3) c...

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Autores principales: Lucchi, Chiara, Codeluppi, Alessandro, Filaferro, Monica, Vitale, Giovanni, Rustichelli, Cecilia, Avallone, Rossella, Mandrioli, Jessica, Biagini, Giuseppe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10135967/
https://www.ncbi.nlm.nih.gov/pubmed/37107338
http://dx.doi.org/10.3390/antiox12040963
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author Lucchi, Chiara
Codeluppi, Alessandro
Filaferro, Monica
Vitale, Giovanni
Rustichelli, Cecilia
Avallone, Rossella
Mandrioli, Jessica
Biagini, Giuseppe
author_facet Lucchi, Chiara
Codeluppi, Alessandro
Filaferro, Monica
Vitale, Giovanni
Rustichelli, Cecilia
Avallone, Rossella
Mandrioli, Jessica
Biagini, Giuseppe
author_sort Lucchi, Chiara
collection PubMed
description We obtained evidence that mouse BV2 microglia synthesize neurosteroids dynamically to modify neurosteroid levels in response to oxidative damage caused by rotenone. Here, we evaluated whether neurosteroids could be produced and altered in response to rotenone by the human microglial clone 3 (HMC3) cell line. To this aim, HMC3 cultures were exposed to rotenone (100 nM) and neurosteroids were measured in the culture medium by liquid chromatography with tandem mass spectrometry. Microglia reactivity was evaluated by measuring interleukin 6 (IL-6) levels, whereas cell viability was monitored by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. After 24 h (h), rotenone increased IL-6 and reactive oxygen species levels by approximately +37% over the baseline, without affecting cell viability; however, microglia viability was significantly reduced at 48 h (p < 0.01). These changes were accompanied by the downregulation of several neurosteroids, including pregnenolone, pregnenolone sulfate, 5α-dihydroprogesterone, and pregnanolone, except for allopregnanolone, which instead was remarkably increased (p < 0.05). Interestingly, treatment with exogenous allopregnanolone (1 nM) efficiently prevented the reduction in HMC3 cell viability. In conclusion, this is the first evidence that human microglia can produce allopregnanolone and that this neurosteroid is increasingly released in response to oxidative stress, to tentatively support the microglia’s survival.
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spelling pubmed-101359672023-04-28 Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress Lucchi, Chiara Codeluppi, Alessandro Filaferro, Monica Vitale, Giovanni Rustichelli, Cecilia Avallone, Rossella Mandrioli, Jessica Biagini, Giuseppe Antioxidants (Basel) Article We obtained evidence that mouse BV2 microglia synthesize neurosteroids dynamically to modify neurosteroid levels in response to oxidative damage caused by rotenone. Here, we evaluated whether neurosteroids could be produced and altered in response to rotenone by the human microglial clone 3 (HMC3) cell line. To this aim, HMC3 cultures were exposed to rotenone (100 nM) and neurosteroids were measured in the culture medium by liquid chromatography with tandem mass spectrometry. Microglia reactivity was evaluated by measuring interleukin 6 (IL-6) levels, whereas cell viability was monitored by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. After 24 h (h), rotenone increased IL-6 and reactive oxygen species levels by approximately +37% over the baseline, without affecting cell viability; however, microglia viability was significantly reduced at 48 h (p < 0.01). These changes were accompanied by the downregulation of several neurosteroids, including pregnenolone, pregnenolone sulfate, 5α-dihydroprogesterone, and pregnanolone, except for allopregnanolone, which instead was remarkably increased (p < 0.05). Interestingly, treatment with exogenous allopregnanolone (1 nM) efficiently prevented the reduction in HMC3 cell viability. In conclusion, this is the first evidence that human microglia can produce allopregnanolone and that this neurosteroid is increasingly released in response to oxidative stress, to tentatively support the microglia’s survival. MDPI 2023-04-19 /pmc/articles/PMC10135967/ /pubmed/37107338 http://dx.doi.org/10.3390/antiox12040963 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lucchi, Chiara
Codeluppi, Alessandro
Filaferro, Monica
Vitale, Giovanni
Rustichelli, Cecilia
Avallone, Rossella
Mandrioli, Jessica
Biagini, Giuseppe
Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title_full Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title_fullStr Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title_full_unstemmed Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title_short Human Microglia Synthesize Neurosteroids to Cope with Rotenone-Induced Oxidative Stress
title_sort human microglia synthesize neurosteroids to cope with rotenone-induced oxidative stress
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10135967/
https://www.ncbi.nlm.nih.gov/pubmed/37107338
http://dx.doi.org/10.3390/antiox12040963
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