London, United Kingdom 2013 12 Experimental studies B 16:50 - 17:00 HYDROGEN SULPHIDE REDUCES INFARCT SIZE AND ENHANCES RECOVERY IN A RAT MODEL: PROTECTION MECHANISM INCLUDES NOX-4 DOWNREGU-LATION I. Henriques1, M. Gutiérrez-Fernández2, B. Rodríguez-Frutos3, J. Ramos4, J. Ferro5, E. Díez-Tejedor6 Neurosciences and Cerebrovascular Research Lab IdiPAZ, Champalimaud NC Program, Centro Hospitalar LZC, *Faculdade de Medicina Universidade de Lisboa, Lisboa, POR-TUGAL1, Neurosciences and Cerebrovascular Research Lab IdiPAZ, Neurology Department Hospital Universitario la Paz, Madrid, SPAIN2, Neurosciences and Cerebrovascular Research Lab IdiPAZ, Neurology Department Hospital Universitario la Paz, Madrid, 3, Neurosciences and Cerebrovascular Research Lab IdiPAZ, Neurology Department Hospital Universitario la Paz, Madrid, SPAIN4, Faculdade de Medicina da Universidade de Lisboa, Lisboa, PORTU-GAL5, Neurosciences and Cerebrovascular Research Lab IdiPAZ, Neurology Department Hos-pital Universitario la Paz, Madrid, SPAIN6 Introduction: Hydrogen sulphide (H2S) inhalation induces a hypometabolic state at micromo-lecular concentrations. The underlying mechanism is not yet completely understood, but in-cludes reduction of oxygen consumption and downregulation of apoptotic pathways. We stud-ied the effect of H2S exposure after acute ischemic stroke in a middle cerebral artery occlusion (MCAO) rat model. Methods: Young adult male Sprague-Dawley rats were distributed into 3 groups: 1-Control: Surgery + permanent MCAO; 2 – Treated: Surgery + permanent MCAO + inhalation of 40 ppM H2S; 3- Sham. We analyzed functional outcome (Roger modified scale) and lesion vol-ume (T2, DWI and PWI MRI at 24h and day 14, and H/E). Cell death was accessed by TUNEL expression at border zone infarct. NOX-4, by immunohistochemistry and Western-Blot quanti-ficaction. Rats were sacrificed at 24h or day 14. Results: Treated animals showed better functional outcome both at 24h (**p = 0, 0044) and day 14 (***p = 0, 0009) and less infarct size at day 14, both in MRI (*p= 0, 0383) and in H/E (*p= 0.0266). Treated animals showed a decrease in TUNEL labeling at border zone infarct (*p= 0, 0307) and a downregulation of NOX-4 expression (* p= 0.0294). Conclusion: Exposure to hydrogen sulphide improved functional recovery after acute MCA occlusion, and reduces infarct size at day 14. Mechanisms include less expression of cell death at the border zone infarct and tissue protection with downregulation of reactive oxygen spe-cies- generating enzyme NOX-4. Probably most H2S benefit occurs before tissue damage. Cerebrovasc Dis 2013; 35 (suppl 3)1-854 79 11 Experimental studies B 16:40 - 16:50 Metabolic profiling of subventricular zones and infarct in a rat model using in vivo mag-netic resonance spectroscopy (MRS) and ex vivo High Resolution Magic Angle Spinning (HRMAS) spectroscopy E. Jiménez-Xarrié1, M. Dávila2, S. Gil-Perotín3, A. Jurado4, A.P. Candiota5, S. Lope-Piedraf-ita6, R. Delgado-Mederos7, J.M. Garcia-Verdugo8, C. Arús9, J. Martí-Fàbregas10 Stroke Unit. Department of Neurology. Hospital de la Santa Creu i Sant Pau., Barcelona, SPAIN1,Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universi-tat Autònoma de Barcelona. CIBER-BBN.IBB, Cerdanyola del Vallès, SPAIN2, Department of Comparative Neurobiology,Instituto Cavanilles de Biodiversidad y Biologia Evolutiva, Universidad de Valencia. CIBERNED., Cerdanyola del Vallès, SPAIN3, Department of Com-parative Neurobiology,Instituto Cavanilles de Biodiversidad y Biologia Evolutiva, Univer-sidad de Valencia., Valencia, SPAIN4, CIBER-BBN. Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universitat Autònoma de Barcelona. IBB., Cerdanyola del Vallès, SPAIN5, Servei de Ressonància Magnètica Nuclear. Universitat Autònoma de Barce-lona. CIBER-BBN., Cerdanyola del Vallès, SPAIN6, Stroke Unit, Department of Neurology, Hospital de la Santa Creu i Sant Pau., Barcelona, SPAIN7, Department of Comparative Neuro-biology, Instituto Cavanilles de Biodiversidad y Biologia Evolutiva, Universidad de Valencia. CIBERNED., Valencia, SPAIN8, Departament de Bioquímica i Biologia Molecular, Facultat de Biociències, Universitat Autònoma de Barcelona .CIBER-BBN. IBB., Cerdanyola del Vallès, SPAIN9, Stroke Unit, Department of Neurology, Hospital de la Santa Creu i Sant Pau., Barcelona, SPAIN10 BACKGROUND: In vivo MRS and ex vivo HRMAS can be used to detect metabolome chang-es in preclinical models. The main advantage of HRMAS is that higher spectral resolution can be achieved. The aim of this study was to analyze the metabolome in the infarct and its adja-cent subventricular zones (SVZ), where changes in the proliferation of neural stem cells are ex-pected, using MRS and HRMAS techniques. METHODS: Sprague-Dawley rats were subjected to 90-minutes MCAO. MRS at 7T was per-formed in non-ischemic rats (n=6), at 1 day (n=6) and at 7 days (n=6) post-stroke. At each time point, half of the animals (n=3) were sacrificed to perform immunohistochemistry in order to detect proliferation in SVZ (Ki67+) and apoptosis (TUNEL+) in the infarct. The remaining ani-mals (n=3) were sacrificed by focused microwave irradiation and their brains were analyzed by ex vivo HRMAS at 9.4T. Unit length normalized peak heights were used for analysis. Statisti-cal differences were assessed with Student’s t-test. RESULTS: Significant metabolome changes were found in the infarct compared to the contra-lateral zone, e.g. apoptotic mobile lipids (2.80 ppm) (up to 5.3 fold-change; p<0.01) at 7 days post-stroke in MRS and also in HRMAS. Accordingly, immunohistochemistry revealed higher apoptosis in the lesion at 7 days compared to the contralateral zone (506 fold-change; p<0.01). Significant increases in the proliferation of cells in the ipsilateral SVZ compared to the contra-lateral SVZ at 7 days (1.4 fold-change; p=0.02) were detected by immunohistochemistry. In vivo MRS did not detect significant changes in the metabolome pattern of the SVZ upon stroke, but ex vivo HRMAS detected an increase in lactate (1.35-1.33 ppm) in ipsilateral (5.86 fold-change; p<0.01) and contralateral SVZ (2.43 fold-change; p=0.02) at 1 day post-stroke com-pared to non-ischemic rats. CONCLUSION: Combining MRS and HRMAS can significantly improve the metabolome characterization of the post-stroke brain.
Karger_ESC London_2013
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