PMID- 31293389 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220409 IS - 1662-5102 (Print) IS - 1662-5102 (Electronic) IS - 1662-5102 (Linking) VI - 13 DP - 2019 TI - Combined Cyclosporin A and Hypothermia Treatment Inhibits Activation of BV-2 Microglia but Induces an Inflammatory Response in an Ischemia/Reperfusion Hippocampal Slice Culture Model. PG - 273 LID - 10.3389/fncel.2019.00273 [doi] LID - 273 AB - INTRODUCTION: Hypothermia attenuates cerebral ischemia-induced neuronal cell death associated with neuroinflammation. The calcineurin inhibitor cyclosporin A (CsA) has been shown to be neuroprotective by minimizing activation of inflammatory pathways. Therefore, we investigated whether the combination of hypothermia and treatment with CsA has neuroprotective effects in an oxygen-glucose deprivation/reperfusion (OGD/R) injury model in neuronal and BV-2 microglia monocultures, as well as in an organotypic hippocampal slice culture (OHSC). METHODS: Murine primary neurons, BV-2 microglia, and OHSC were pretreated with CsA and exposed to 1 h OGD (0.2% O(2)) followed by reperfusion at normothermia (37 degrees C) or hypothermia (33.5 degrees C). Cytotoxicity was measured by lactate dehydrogenase and glutamate releases. Damage-associated molecular patterns (DAMPs) high mobility group box 1 (HMGB1), heat shock protein 70 (Hsp70), and cold-inducible RNA-binding protein (CIRBP) were detected in cultured supernatant by western blot analysis. Interleukin-6 (IL-6), Interleukin-1alpha and -1beta (IL-1alpha/IL1-beta), tumor necrosis factor-alpha (TNF-alpha), monocyte chemotactic protein 1 (MCP1), inducible nitric oxide synthase (iNOS), glia activation factors ionized calcium-binding adapter molecule 1 (Iba1), and transforming growth factor beta1 (TGF-beta1) gene expressions were analyzed by RT-qPCR. RESULTS: Exposure to OGD plus 10 muM CsA was sufficient to induce necrotic cell death and subsequent release of DAMPs in neurons but not BV-2 microglia. Moreover, OGD/R-induced secondary injury was also observed only in the neurons, which was not attenuated by cooling and no increased toxicity by CsA was observed. BV-2 microglia were not sensitive to OGD/R-induced injury but were susceptible to CsA-induced toxicity in a dose dependent manner, which was minimized by hypothermia. CsA attenuated IL-1beta and Iba1 expressions in BV-2 microglia exposed to OGD/R. Hypothermia reduced IL-1beta and iNOS expressions but induced TNF-alpha and Iba1 expressions in the microglia. However, these observations did not translate to the ex vivo OHCS model, as general high expressions of most cytokines investigated were observed. CONCLUSION: Treatment with CsA has neurotoxic effects on primary neurons exposed to OGD but could inhibit BV-2 microglia activation. However, CsA and hypothermia treatment after ischemia/reperfusion injury results in cytotoxic neuroinflammation in the complex ex vivo OHSC. FAU - Wowro, Sylvia J AU - Wowro SJ AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. AD - Charite - Universitatsmedizin Berlin, Corporate Member of Freie Universitat Berlin, Humboldt-Universitat zu Berlin, and Berlin Institute of Health, Berlin, Germany. FAU - Tong, Giang AU - Tong G AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. FAU - Krech, Jana AU - Krech J AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. FAU - Rolfs, Nele AU - Rolfs N AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. FAU - Berger, Felix AU - Berger F AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. AD - Charite - Universitatsmedizin Berlin, Corporate Member of Freie Universitat Berlin, Humboldt-Universitat zu Berlin, and Berlin Institute of Health, Berlin, Germany. FAU - Schmitt, Katharina R L AU - Schmitt KRL AD - Department of Congenital Heart Disease/Pediatric Cardiology, Universitares Herzzentrum Berlin - Medical Heart Center of Charite and German Heart Institute Berlin, Berlin, Germany. AD - Charite - Universitatsmedizin Berlin, Corporate Member of Freie Universitat Berlin, Humboldt-Universitat zu Berlin, and Berlin Institute of Health, Berlin, Germany. LA - eng PT - Journal Article DEP - 20190625 PL - Switzerland TA - Front Cell Neurosci JT - Frontiers in cellular neuroscience JID - 101477935 PMC - PMC6603137 OTO - NOTNLM OT - BV-2 microglia OT - DAMPs OT - cyclosporin A OT - hypothermia OT - inflammation OT - organotypic hippocampal slice culture OT - oxygen-glucose deprivation/reperfusion OT - primary neuron EDAT- 2019/07/12 06:00 MHDA- 2019/07/12 06:01 PMCR- 2019/01/01 CRDT- 2019/07/12 06:00 PHST- 2019/03/27 00:00 [received] PHST- 2019/06/05 00:00 [accepted] PHST- 2019/07/12 06:00 [entrez] PHST- 2019/07/12 06:00 [pubmed] PHST- 2019/07/12 06:01 [medline] PHST- 2019/01/01 00:00 [pmc-release] AID - 10.3389/fncel.2019.00273 [doi] PST - epublish SO - Front Cell Neurosci. 2019 Jun 25;13:273. doi: 10.3389/fncel.2019.00273. eCollection 2019.