Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats
Abstract Background and purpose The only validated treatment to prevent brain damage associated with hypoxia–ischemia (HI) encephalopathy of the newborn is controlled hypothermia with limited benefits. Additional putative neuroprotective drug candidates include sildenafil citrate, a phosphodiesteras...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-01-01
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| Series: | Journal of Neuroinflammation |
| Online Access: | https://doi.org/10.1186/s12974-024-03022-w |
| _version_ | 1797276394123689984 |
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| author | Pansiot Julien Manuela Zinni Natacha Bonnel Marina El Kamouh Felipe Odorcyk Lea Peters Emilie-Fleur Gautier Marjorie Leduc Cédric Broussard Olivier Baud |
| author_facet | Pansiot Julien Manuela Zinni Natacha Bonnel Marina El Kamouh Felipe Odorcyk Lea Peters Emilie-Fleur Gautier Marjorie Leduc Cédric Broussard Olivier Baud |
| author_sort | Pansiot Julien |
| collection | DOAJ |
| description | Abstract Background and purpose The only validated treatment to prevent brain damage associated with hypoxia–ischemia (HI) encephalopathy of the newborn is controlled hypothermia with limited benefits. Additional putative neuroprotective drug candidates include sildenafil citrate, a phosphodiesterase-type 5 inhibitor. The main objective of this preclinical study is to assess its ability to reduce HI-induced neuroinflammation, in particular through its potential effect on microglial activation. Methods HI was induced in P10 Sprague–Dawley rats by unilateral carotid permanent artery occlusion and hypoxia (HI) and treated by either hypothermia (HT) alone, Sildenafil (Sild) alone or combined treatment (SildHT). Lesion size and glial activation were analyzed by immunohistochemistry, qRT-PCR, and proteomic analyses performed at P13. Results None of the treatments was associated with a significant early reduction in lesion size 72h after HI, despite significant changes in tissue loss distribution. Significant reductions in both Iba1 + (within the ipsilateral hemisphere) and GFAP + cells (within the ipsilateral hippocampus) were observed in SildHT group, but not in the other treatment groups. In microglia-sorted cells, pro-inflammatory markers, i.e. Il1b, Il6, Nos2, and CD86 were significantly downregulated in SildHT treatment group only. These changes were restricted to the ipsilateral hemisphere, were not evidenced in sorted astrocytes, and were not sex dependent. Proteomic analyses in sorted microglia refined the pro-inflammatory effect of HI and confirmed a biologically relevant impact of SildHT on specific molecular pathways including genes related to neutrophilic functions. Conclusions Our findings suggest that Sildenafil combined with controlled hypothermia produces maximum effect in mitigating microglial activation induced by HI through complex proteomic regulation. The reduction of neuroinflammation induced by Sildenafil may represent an interesting therapeutic strategy for neonatal neuroprotection. |
| first_indexed | 2024-03-07T15:27:40Z |
| format | Article |
| id | doaj.art-51b6c995106742d99a58c08c35e8a0b9 |
| institution | Directory Open Access Journal |
| issn | 1742-2094 |
| language | English |
| last_indexed | 2024-03-07T15:27:40Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Neuroinflammation |
| spelling | doaj.art-51b6c995106742d99a58c08c35e8a0b92024-03-05T16:36:55ZengBMCJournal of Neuroinflammation1742-20942024-01-0121111310.1186/s12974-024-03022-wSynergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in ratsPansiot Julien0Manuela Zinni1Natacha Bonnel2Marina El Kamouh3Felipe Odorcyk4Lea Peters5Emilie-Fleur Gautier6Marjorie Leduc7Cédric Broussard8Olivier Baud9Inserm UMR1141 NeuroDiderot, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéInstitut Cochin, Proteom’IC Facility, INSERM, CNRS, Université Paris CitéInstitut Cochin, Proteom’IC Facility, INSERM, CNRS, Université Paris CitéInstitut Cochin, Proteom’IC Facility, INSERM, CNRS, Université Paris CitéInserm UMR1141 NeuroDiderot, Université Paris CitéAbstract Background and purpose The only validated treatment to prevent brain damage associated with hypoxia–ischemia (HI) encephalopathy of the newborn is controlled hypothermia with limited benefits. Additional putative neuroprotective drug candidates include sildenafil citrate, a phosphodiesterase-type 5 inhibitor. The main objective of this preclinical study is to assess its ability to reduce HI-induced neuroinflammation, in particular through its potential effect on microglial activation. Methods HI was induced in P10 Sprague–Dawley rats by unilateral carotid permanent artery occlusion and hypoxia (HI) and treated by either hypothermia (HT) alone, Sildenafil (Sild) alone or combined treatment (SildHT). Lesion size and glial activation were analyzed by immunohistochemistry, qRT-PCR, and proteomic analyses performed at P13. Results None of the treatments was associated with a significant early reduction in lesion size 72h after HI, despite significant changes in tissue loss distribution. Significant reductions in both Iba1 + (within the ipsilateral hemisphere) and GFAP + cells (within the ipsilateral hippocampus) were observed in SildHT group, but not in the other treatment groups. In microglia-sorted cells, pro-inflammatory markers, i.e. Il1b, Il6, Nos2, and CD86 were significantly downregulated in SildHT treatment group only. These changes were restricted to the ipsilateral hemisphere, were not evidenced in sorted astrocytes, and were not sex dependent. Proteomic analyses in sorted microglia refined the pro-inflammatory effect of HI and confirmed a biologically relevant impact of SildHT on specific molecular pathways including genes related to neutrophilic functions. Conclusions Our findings suggest that Sildenafil combined with controlled hypothermia produces maximum effect in mitigating microglial activation induced by HI through complex proteomic regulation. The reduction of neuroinflammation induced by Sildenafil may represent an interesting therapeutic strategy for neonatal neuroprotection.https://doi.org/10.1186/s12974-024-03022-w |
| spellingShingle | Pansiot Julien Manuela Zinni Natacha Bonnel Marina El Kamouh Felipe Odorcyk Lea Peters Emilie-Fleur Gautier Marjorie Leduc Cédric Broussard Olivier Baud Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats Journal of Neuroinflammation |
| title | Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats |
| title_full | Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats |
| title_fullStr | Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats |
| title_full_unstemmed | Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats |
| title_short | Synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia–ischemia in rats |
| title_sort | synergistic effect of sildenafil combined with controlled hypothermia to alleviate microglial activation after neonatal hypoxia ischemia in rats |
| url | https://doi.org/10.1186/s12974-024-03022-w |
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