RESUMEN
AIMS: To explore cardiac structural and functional parameters and myocardial sensitivity to ischemia in a rat model of chronic arthritis, pristane-induced arthritis (PIA), and to investigate the effects of a running exercise protocol on cardiac disorders related to rheumatoid arthritis (RA). MAIN METHODS: 3 groups of male Dark Agouti rats were formed: Controls, PIA and PIA-Exercise. The PIA-Exercise group was subjected to an individualized treadmill running protocol during the remission phase. At acute and chronic phases of PIA, cardiac structure was analyzed by histology. Cardiac function was explored in isolated hearts to measure left ventricular developed pressure (LVDP), cardiac compliance and infarct size before and after ischemia/reperfusion. Cardiac inflammation was evaluated through VCAM-1 mRNA expression by RT-qPCR. Plasma irisin levels were measured by ELISA. KEY FINDINGS: PIA rats exhibited myocardial hypertrophy fibrosis and inflammation at the 2 inflammatory phases of the model. At chronic phase only, LVDP and cardiac compliance were lower in PIA compared to controls. As compared to sedentary PIA, exercise did not change cardiac function but reduced fibrosis, inflammation, infarct size, and arthritis severity and increased irisin levels. Cardiac inflammation positively correlated with fibrosis, while irisin levels negatively correlated with cardiac inflammation and fibrosis. SIGNIFICANCE: In the PIA model that recapitulated most cardiac disorders of RA, a daily program of treadmill running alleviated cardiac fibrosis and inflammation and improved resistance to ischemia. These data provide arguments to promote the practice of exercise in RA patients for cardiac diseases prevention.
Asunto(s)
Artritis Experimental , Artritis Reumatoide , Cardiopatías , Terpenos , Humanos , Ratas , Masculino , Animales , Artritis Experimental/metabolismo , Fibronectinas/efectos adversos , Inflamación , Artritis Reumatoide/metabolismo , Isquemia , Infarto , FibrosisRESUMEN
AIM: This study explored the systemic vascular effects of local cryotherapy with a focus on endothelial changes and arterial inflammation in the model of rat adjuvant-induced arthritis (AIA). METHODS: Cryotherapy was applied twice a day on hind paws of AIA rats from the onset of arthritis to the acute inflammatory phase. Endothelial activation was studied in the aorta by measuring the mRNA levels of chemokines (CXCL-1, MCP-1 (CCL-2), MIP-1α (CCL-3)) and adhesion molecules (ICAM-1, VCAM-1) by qRT-PCR. Endothelial dysfunction was measured in isolated aortic and mesenteric rings. Aortic inflammation was evaluated via the mRNA expression of pro-inflammatory cytokines (TNF-α, IL-6) by qRT-PCR and leucocyte infiltration analysis (flow cytometry). Plasma levels of TNF-α, IL-6, IL-1ß, IL-17A, and osteoprotegerin (OPG) were measured using Multiplex/ELISA. RESULTS: AIA was associated with an increased aortic expression of CXCL-1 and ICAM-1 as well as an infiltration of leucocytes and increased mRNA expression of IL-6, IL-1ß, and TNF-α. Local cryotherapy, which decreased arthritis score and structural damages, reduced aortic mRNA expression of CXCL-1, IL-6, IL-1ß, and TNF-α, as well as aortic infiltration of leucocytes (T lymphocytes, monocytes/macrophages, neutrophils) and improved acetylcholine-induced vasorelaxation in the aorta and mesenteric arteries. Plasma levels of IL-17A and OPG were significantly reduced by cryotherapy, while the number of circulating leucocytes was not. IL-17A levels positively correlated with endothelial activation and dysfunction. CONCLUSION: In the AIA model, local cryotherapy reduced systemic endothelial activation, immune cell infiltration, and endothelial dysfunction. Mechanistically, the reduction of circulating levels of IL-17A appears as the possible link between joint cooling and the remote vascular effects.
Asunto(s)
Artritis Experimental , Molécula 1 de Adhesión Intercelular , Animales , Crioterapia , Inflamación , Interleucina-17 , Interleucina-6 , ARN Mensajero , Ratas , Ratas Endogámicas Lew , Factor de Necrosis Tumoral alfaRESUMEN
Little is known on the cerebrovascular BDNF (brain-derived neurotrophic factor)/TrkB (tropomyosin related kinase B) pathway. This study investigated the contribution of endogenous endothelial BDNF to the control of vascular tone of rat middle cerebral artery (MCA) and the capacity of exogenous agonist of TrkB receptors to induce their relaxation. Endothelial cells constitutively expressed both BDNF and activated TrkB receptors. Supporting endothelial BDNF as an autocrine regulator of basal myogenic tone, incubation of MCA with the TrkB antagonist cyclotraxin B induced contraction as observed with incubation in the presence of inhibitors of nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF) production. Exposure of MCA with the TrkB agonist LM22A-4 that increased expression of TrkB receptors phosphorylated at tyrosine 816 induced relaxation of preconstricted MCA (EC50 6.7 × 10-8 mol/L) as efficiently than acetylcholine (EC50 5.3 × 10-8 mol/L). Finally, endothelium removal, exposure to a TrkB antagonist or to inhibitors of NO and EDHF production prevented the relaxant effect of LM22A-4. In conclusion, our study identified endothelial BDNF as a new autocrine regulator of vascular tone of MCA, thus making the endothelial BDNF/TrkB pathway an attractive target for strategies aiming to improve blood supply to the brain.
Asunto(s)
Células Endoteliales , Receptor trkB , Animales , Factores Biológicos , Células Endoteliales/metabolismo , Endotelio/metabolismo , Arteria Cerebral Media/metabolismo , Ratas , Receptor trkB/metabolismoRESUMEN
While brain-derived neurotrophic factor (BDNF) was previously reported to induce relaxation of conduit artery, whether the BDNF/TrkB (tropomyosin-related kinase) pathway is involved in the tone control of resistance arteries is not known. This study investigated TrkB receptors levels/localization and the vasomotor effect of the TrkB receptor agonist LM22A-4 in isolated third-order mesenteric arteries from rats. Immunostaining revealed the presence of both full-length and truncated TrkB receptors, especially at the endothelial level. By using wire myography, LM22A-4 induced vascular relaxation that was significantly decreased by cyclotraxin B as a non-competitive TrkB antagonist and fully prevented by endothelium removal. Inhibitors of NO, EDHF, PGI2 production and the PI3K/Akt pathways separately reduced LM22A-4 induced-relaxation. By contrast, inhibition of Raf/MEK, PLCγ and CaM/CaMKII pathways did not change the relaxant effect of LM22A-4. Interestingly, BDNF also induced an endothelium and TrkB-dependent relaxation. These results indicate that endothelial TrkB activation results in the relaxation of resistance vessels via PI3K/Akt-induced eNOS phosphorylation and production of EDHF and PGI2. These data are consistent with the contribution of the endothelial BDNF/TrkB pathway to the regulation of peripheral vascular tone. They also validate the use of LM22A-4 as a reliable pharmacological agent for studying the vascular effect of BDNF.
Asunto(s)
Benzamidas/farmacología , Células Endoteliales/efectos de los fármacos , Arterias Mesentéricas/efectos de los fármacos , Receptor trkB/agonistas , Resistencia Vascular/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacología , Animales , Factores Biológicos/metabolismo , Relación Dosis-Respuesta a Droga , Células Endoteliales/metabolismo , Epoprostenol/metabolismo , Técnicas In Vitro , Masculino , Arterias Mesentéricas/metabolismo , Miografía , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo III/metabolismo , Fosfatidilinositol 3-Quinasa/metabolismo , Fosforilación , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas Wistar , Receptor trkB/metabolismo , Transducción de Señal/efectos de los fármacosRESUMEN
To determine the effect of glucocorticoids (GCs) on endothelial dysfunction (ED) and on traditional cardiovascular (CV) risk factors in the adjuvant-induced arthritis (AIA) rat model. At the first signs of AIA, a high dose (HD) [10 mg/kg/day, intraperitoneally (i.p.), GC-HD] or low dose (LD) (1 mg/kg/day, i.p., GC-LD) of prednisolone was administered for 3 weeks. Endothelial function was studied in aortic rings relaxed with acetylcholine (Ach) with or without inhibitors of nitric oxide synthase (NOS), cyclooxygenase 2 (COX-2), arginase, endothelium derived hyperpolarizing factor (EDHF) and superoxide anions ( O2-°) production. Aortic expression of endothelial NOS (eNOS), Ser1177-phospho-eNOS, COX-2, arginase-2, p22phox and p47phox was evaluated by Western blotting analysis. Arthritis scores, blood pressure, heart rate and blood levels of cytokines, triglycerides, cholesterol and glucose were measured. GC-HD but not GC-LD reduced arthritis score significantly and improved Ach-induced relaxation (P < 0·05). The positive effect of GC-HD resulted from increased NOS activity and EDHF production and decreased COX-2/arginase activities and O2-° production. These functional effects relied upon increased phospho-eNOS expression and decreased COX-2, arginase-2 and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase expression. Despite the lack of effect of GC-LD on ED, it increased NOS and EDHF and down-regulated O2-° pathways but did not change arginase and COX-2 pathways. GC-HD increased triglycerides levels and blood pressure significantly (P < 0·05). Both doses of GCs decreased to the same extent as plasma interleukin (IL)-1ß and tumour necrosis factor (TNF)-α levels (P < 0·05). Our data demonstrated that subchronic treatment with prednisolone improved endothelial function in AIA via pleiotropic effects on endothelial pathways. These effects occurred independently of the deleterious cardiometabolic effects and the impact of prednisolone on systemic inflammation.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Artritis Reumatoide/tratamiento farmacológico , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/fisiología , Glucocorticoides/administración & dosificación , Prednisolona/administración & dosificación , Acetilcolina/farmacología , Animales , Aorta/fisiopatología , Arginasa/farmacología , Artritis , Artritis Reumatoide/fisiopatología , Factores Biológicos/metabolismo , Glucemia/análisis , Presión Sanguínea , Enfermedades Cardiovasculares/diagnóstico , Colesterol/sangre , Ciclooxigenasa 2/farmacología , Citocinas/sangre , Frecuencia Cardíaca , Masculino , Distribución Aleatoria , Ratas , Superóxidos/metabolismo , Triglicéridos/sangreRESUMEN
AIM: Evidence that brain-derived neurotrophic factor (BDNF), a neurotrophin largely involved in cognition, is expressed by cerebral endothelial cells led us to explore in rats the contribution of the cerebral microvasculature to BDNF found in brain tissue and the link between cerebrovascular nitric oxide (NO) and BDNF production. METHODS: Brain BDNF protein levels were measured before and after in situ removal of the cerebral endothelium that was achieved by brain perfusion with a 0.2% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propane sulphonate) solution. BDNF protein and mRNA levels as well as levels of endothelial NO synthase phosphorylated at serine 1177 (P-eNOSser1177 ) were measured in cerebral microvessel-enriched fractions. These fractions were also exposed to glycerol trinitrate. Hypertension (spontaneously hypertensive rats) and physical exercise training were used as experimental approaches to modulate cerebrovascular endothelial NO production. RESULTS: CHAPS perfusion resulted in a marked decrease in brain BDNF levels. Hypertension decreased and exercise increased P-eNOSser1177 and BDNF protein levels. However, BDNF mRNA levels that were increased by exercise did not change after hypertension. Finally, in vitro exposure of cerebral microvessel-enriched fractions to glycerol trinitrate enhanced BDNF production. CONCLUSION: These data reveal that BDNF levels measured in brain homogenates correspond for a large part to BDNF present in cerebral endothelial cells and that cerebrovascular BDNF production is dependent on cerebrovascular endothelial eNOS activity. They provide a paradigm shift in the cellular source of brain BDNF and suggest a new approach to improve our understanding of the link between endothelial function and cognition.
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Factor Neurotrófico Derivado del Encéfalo/metabolismo , Encéfalo/metabolismo , Células Endoteliales/metabolismo , Microvasos/metabolismo , Animales , Western Blotting , Encéfalo/irrigación sanguínea , Inmunohistoquímica , Óxido Nítrico/metabolismo , Reacción en Cadena de la Polimerasa , Ratas , Ratas Endogámicas SHR , Ratas WistarRESUMEN
The enzyme arginase catalyses the divalent cation dependent hydrolysis of L-arginine to produce L-ornithine and urea. Two isoforms of arginases have been identified in mammalian (including human) cells. Moreover, some infectious pathogens (e.g. Leishmania) synthesize their own arginase. Work over the last decades has revealed that elevated arginase activity both decreases cellular availability in nitric oxide (NO) by competing with NO synthases (NOS) and increases concentration in L-ornithine, a precursor in the biosynthesis of polyamines which are important for cell differentiation and proliferation. From these data emerged the concept that selective arginase inhibitors might be a valuable strategy for treatment of various diseases associated with decreased NO and/or increased polyamines production. Consistent with this, recent research provides compelling evidence supporting the beneficial effects of arginase inhibitors in cardiovascular diseases (hypertension, ischemia reperfusion injury, atherosclerosis, diabetes mellitus), asthma, cancer, immunologically-mediated diseases or leishmaniasis. Despite active programs to identify potent arginase inhibitors, effective chemical compounds with reliable pharmacokinetics and toxicological properties are rare. The present review summarizes available data on the discovery of new arginase inhibitors from natural origin. Current knowledge on plant-derived compounds or extracts with arginase inhibitory properties as well as available data on structure-activity relationship (SAR) will be presented. Lastly, the present review will open up new prospects in order to improve the discovery of novel arginase inhibitors from natural sources.
Asunto(s)
Arginasa/antagonistas & inhibidores , Inhibidores Enzimáticos/química , Animales , Arginasa/metabolismo , Supervivencia Celular/efectos de los fármacos , Inhibidores Enzimáticos/metabolismo , Humanos , Leishmania/enzimología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa/metabolismo , Extractos Vegetales/química , Extractos Vegetales/farmacología , Plantas/química , Plantas/metabolismo , Proteínas Protozoarias/antagonistas & inhibidores , Proteínas Protozoarias/metabolismo , Relación Estructura-ActividadRESUMEN
Our group demonstrated recently that arginase I inhibition reduces endothelial dysfunction and blood pressure rising in spontaneously hypertensive rats [C. Demougeot, A. Prigent-Tessier, C. Marie, A. Berthelot, J. Hypertens. 23 (2005) 971; C. Demougeot, A. Prigent-Tessier, T. Bagnost, C. Andre, Y. Guillaume, M. Bouhaddi, C. Marie, A. Berthelot, Life Sci. 80 (2007) 1128]. This discovery opens interesting perspectives in the development of new drugs against hypertension. As well, in a previous paper [T. Bagnost, Y.C. Guillaume, M. Thomassin, J.F. Robert, A. Berthelot, A. Xicluna, C. Andre, J. Chromatogr. B: Analyt. Technol. Biomed. Life Sci. 856 (2007) 113], a novel biochromatographic column was developed in our laboratory for studying the binding of N(omega)-hydroxy-nor-l-arginine (nor-NOHA), an arginase inhibitor, with this enzyme. In this manuscript, using this novel biochromatographic concept, the effect of magnesium chloride on the nor-NOHA/arginase binding was analyzed for the first time. This study demonstrated that the salt ions interacted with arginase and played a great role in the nor-NOHA/arginase association. For a salt concentration (x) in the medium less than 3mM, the nor-NOHA/arginase binding decreased with x due to a decrease of the charge-charge interactions between nor-NOHA and its arginase binding site. Above 3mM of salt in the medium, the affinity of nor-NOHA to arginase increased slightly with x because the net number of ions (n) (Mg(2+) or Cl(-)) released or bound upon complex formation is low. As well, it was clearly demonstrated, that above 3 mM the n value depend on the salt concentration in the bulk solvent and was approximately nil for x=12 mM. This dependence was due to a gradual and conformational change of the arginase enzyme which around 12 mM adopted a less flexible structure; its binding site was thus less accessible to nor-NOHA and nor-NOHA-arginase association decreased slightly.
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Arginasa/metabolismo , Arginina/análogos & derivados , Cloruro de Magnesio/farmacología , Arginina/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Unión Proteica/efectos de los fármacos , TermodinámicaRESUMEN
Phytoestrogens, naturally occurring plant compounds having oestrogenic and/or anti-oestrogenic activity, are present in many human foodstuffs including hop. Moderate intakes of isoflavonoid phytoestrogens have been associated with a reduction in cardiovascular diseases incidence. So, it is possible that hop (Humulus Lupulus L.) might similarly contribute to the reported health-beneficial effects of moderate beer consumption. Thus, the purpose of this study was to investigate in vitro effects of aqueous hop extract on thoracic vascular reactivity in Sprague Dawley male and female rats. Endothelium-intact thoracic arterial rings from male rats (MALE, n=8), sham-ovariectomized (Sham OVX) female (n=8) and ovariectomized (OVX) female rats (n=8) were used. We assessed the relaxation induced by aqueous hop extract (10(-9), 10(-2)g/l) in aortic rings precontracted with norepinephrine (10(-7)M), in the absence or in the presence of l-NAME (10(-4)M), indomethacin (10(-5)M), thapsigargin (10(-4)M), iberiotoxin (3.10(-8)M), apamin (3.10(-8)M) and TEA (3.10(-4)M). Aqueous hop extract induced relaxation of endothelium-intact thoracic arterial rings in MALE and Sham OVX rats, whereas a weak effect was observed in OVX rats. This vasorelaxation was strongly inhibited in presence of l-NAME, indomethacin and thapsigargin. These data indicated that aqueous hop extract-induced vasodilation, in male and intact female rats, is mediated by NOS activation, cyclooxygenase products and Ca(2+) pathways. Moreover, our results suggested that effect of hop in enhancing vascular reactivity was independent of gender but strongly related to hormonal status.
Asunto(s)
Aorta Torácica/efectos de los fármacos , Humulus/química , Extractos Vegetales/farmacología , Vasodilatación/efectos de los fármacos , Vasodilatadores/farmacología , Animales , Inhibidores Enzimáticos/farmacología , Femenino , Técnicas In Vitro , Indometacina/farmacología , Masculino , NG-Nitroarginina Metil Éster/farmacología , Norepinefrina/farmacología , Ovariectomía , Bloqueadores de los Canales de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Factores Sexuales , Tapsigargina/farmacología , Vasoconstricción/efectos de los fármacos , Vasoconstrictores/farmacologíaRESUMEN
The purpose of this study was to investigate the effects of a 8-week of swim training on total plasma homocysteine and cysteine levels in 16 male Sprague-Dawley rats aged 17 weeks. We also evaluated the activity of hepatic cystathionine beta-synthase (CBS), an enzyme involved in the metabolism of Hcy, the concentration of plasma glutathione, taurine, and a fraction of vitamin B6: the pyridoxal 5-phosphate (PLP). After one week of acclimatization, rats were randomly divided into two groups: 8 non-trained (NTR) and 8 trained rats (TR). Following the training period, body weight gain was lower in TR than in NTR. Plasma homocysteine did not differ among groups while significantly lower plasma cysteine and taurine levels were found in TR (157.83 +/- 8.6 micromol/L; 133.01 +/- 9.32 micromol/L; P < 0.05) compared with data of NTR (176.19 +/- 4.9 micromol/L; 162.57 +/- 8.16 micromol/L; P < 0.05). No significant changes in hepatic CBS activity were observed in TR compared with NTR. Moreover, values for plasma glutathione and PLP concentrations were not affected by training.These results indicate that training reduces plasma cysteine and taurine levels whereas it does not modify other studied parameters. Thus, physical training may regulate cysteine metabolism.
Asunto(s)
Cisteína/sangre , Homocisteína/sangre , Condicionamiento Físico Animal , Natación/fisiología , Animales , Cistationina betasintasa/metabolismo , Glutatión/sangre , Hígado/enzimología , Masculino , Condicionamiento Físico Animal/métodos , Fosfato de Piridoxal/sangre , Ratas , Ratas Sprague-Dawley , Taurina/sangreRESUMEN
We examined the efficacy of the liposoluble iron chelator 2,2'-dipyridyl (DP) in reducing histological damage in rats submitted to cerebral ischemia and the mechanisms involved in the potential cytoprotection. For this purpose, DP (20 mg/kg, i.p.) was administered 15 min before and 1 h after induction of cortical photothrombotic vascular occlusion in rat. Histological studies were performed to assess infarct volume (at days 1 and 3 postischemia) and astromicroglial activation (at day 3 postischemia). Damage to endothelial and neuronal cells was evaluated at day 1 postischemia by quantitative measurements of Evans Blue extravasation and N-acetylaspartate levels, respectively. Cerebral blood flow was recorded in the ischemic core by laser-Doppler flowmetry within the 15 min to 2 h period after photothrombosis. At 4-h postischemia, radical oxygen species (ROS) production was evaluated by measuring brain glutathione concentrations. The cortical expression of the proteins heme oxygenase-1 (HO-1) and hypoxia-inducible factor-1alpha (HIF-1alpha) was analyzed by Western blotting at day 1 postischemia. Infarct volume and ischemic damage to endothelial and neuronal cells were significantly reduced by DP treatment. This cytoprotection was associated with a reduction in ROS production, perfusion deficits, and astrocytic activation. DP treatment also resulted in significant changes in HO-1 (+100%) and HIF-1alpha (-50%) protein expression at the level of the ischemic core. These results report the efficacy of the liposoluble iron chelator DP in reducing histological damage induced by permanent focal ischemia.
Asunto(s)
2,2'-Dipiridil/uso terapéutico , Isquemia Encefálica/tratamiento farmacológico , Quelantes del Hierro/uso terapéutico , Accidente Cerebrovascular/tratamiento farmacológico , 2,2'-Dipiridil/química , Animales , Astrocitos/efectos de los fármacos , Astrocitos/patología , Isquemia Encefálica/patología , Supervivencia Celular/efectos de los fármacos , Corteza Cerebral/patología , Infarto Cerebral/patología , Circulación Cerebrovascular/efectos de los fármacos , Proteínas de Unión al ADN/metabolismo , Glutatión/metabolismo , Hemo Oxigenasa (Desciclizante)/biosíntesis , Hemo-Oxigenasa 1 , Factor 1 Inducible por Hipoxia , Subunidad alfa del Factor 1 Inducible por Hipoxia , Quelantes del Hierro/química , Rayos Láser , Masculino , Neuroglía/efectos de los fármacos , Neuroglía/patología , Neuronas/patología , Proteínas Nucleares/metabolismo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Accidente Cerebrovascular/patología , Trombosis/tratamiento farmacológico , Trombosis/patología , Factores de Transcripción/metabolismoRESUMEN
Rats fed with low (0.015%), normal (0.08%) or high (0.32%) magnesium (Mg) diet for 5-6 weeks were subjected to photothrombosis-induced infarction. As compared to normal diet, Mg deprivation increased by 45% infarct volume at 24 h after photothrombosis but did not modify the lesion at 4 h after photothrombosis. Mg supplementation did not protect from infarction whatever the time point examined. No differences in pre-ischemic systolic blood pressure and glycemia as well as in post-ischemic kaliemia, calcemia and plasma antioxidant activity were observed between groups. However, plasma total Mg level correlated with plasma antioxidant activity at 4 h after photothrombosis. These results demonstrate that brains from Mg deficient rats are more susceptible to permanent focal ischemia than rats fed with normal or high Mg diet.
Asunto(s)
Isquemia Encefálica/sangre , Deficiencia de Magnesio/sangre , Magnesio/sangre , Accidente Cerebrovascular/sangre , Animales , Isquemia Encefálica/dietoterapia , Magnesio/farmacología , Magnesio/uso terapéutico , Masculino , Ratas , Ratas Wistar , Accidente Cerebrovascular/dietoterapiaRESUMEN
Inducible nitric oxide synthase (iNOS) protects heart against ischemia/reperfusion injury. However, it is unknown whether the beneficial effects of iNOS are mediated by the interaction of NO with radical oxygen species (ROS). To address this issue, we examined the effects of liposoluble iron-induced ROS generation in isolated perfused hearts from rats treated with lipopolysaccharide (LPS). LPS administration (10 mg/kg, i.p., 6 h before heart removal) induced iNOS expression and increased NO production as indicated by a 3-fold elevation of nitrite level in coronary effluents relative to control hearts. An enhanced expression of hemeoxygenase 1 protein was also observed in septic hearts compared to control. Iron-induced perfusion and contractile deficits were ameliorated by LPS with more important coronary than myocardial benefits. In iron-loaded hearts, oxidative stress as measured by the 2,3 dihydroxybenzoic acid/salicylic acid concentration ratio in cardiac tissue was 23% lower in septic than in control heart although the difference did not reach significance. In addition, the presence of the NO synthase inhibitor N-nitro-L-arginine in the perfusion medium totally blocked NO production but did not reverse the protective effects of LPS. The results indicate that LPS protects from iron-induced cardiac dysfunction by mechanisms independent on ex vivo NO production and suggest that NO acts as a trigger rather than a direct mediator of the cardioprotective effects of LPS in heart exposed to iron.
Asunto(s)
Corazón/efectos de los fármacos , Hierro/metabolismo , Miocardio/enzimología , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/metabolismo , Animales , Arginina/farmacología , Ventrículos Cardíacos/efectos de los fármacos , Hidroxibenzoatos/metabolismo , Radical Hidroxilo , Immunoblotting , Lipopolisacáridos/metabolismo , Contracción Miocárdica/efectos de los fármacos , Óxido Nítrico/química , Óxido Nítrico Sintasa de Tipo II , Nitritos/metabolismo , Nitroarginina/química , Estrés Oxidativo , Perfusión , Ratas , Ratas Wistar , Daño por Reperfusión/prevención & control , Ácido Salicílico/metabolismo , Factores de TiempoRESUMEN
For a better understanding of the role of iron imbalance in neuropathology, a liposoluble iron complex (ferric hydroxyquinoline, FHQ) was injected into striatum of rats. The effects of two modalities of iron injections on brain damage, hydroxyl radical (*OH) production (assessed by the salicylate method coupled to microdialysis) and tissue reactive iron level (evaluated ex vivo by the propensity of the injected structure for lipid peroxidation) were examined. Rapid injection of FHQ (10 nmoles of 5 mM FHQ pH 3 solution over 1-min period) but not that of corresponding vehicle led to extensive damage associated with increased tissue free iron level in the injected region. Conversely, neither lesion nor free iron accumulation was observed after slow FHQ injection (10 nmoles of a 100 microM FHQ pH 7 solution over 1-h period) as compared to corresponding vehicle injection. Production of *OH was induced by slow FHQ injection but not by rapid FHQ injection, probably as a result of in vivo abolition of iron-induced *OH formation by acid pH. Indeed, rapid injection of FAC pH 7 (ferric ammonium citrate, 5 mM in saline) was associated with *OH formation whereas rapid injection of FAC pH 3 did not. Our results identify the rate of iron delivery to cells as an important determinant of iron toxicity and do not support a major role for extracellular *OH in damage associated with intracerebral iron injection.
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Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Compuestos Férricos/administración & dosificación , Compuestos Férricos/farmacocinética , Hidroxiquinolinas/administración & dosificación , Hidroxiquinolinas/farmacocinética , Hierro/administración & dosificación , Hierro/farmacocinética , Animales , Apomorfina/farmacología , Muerte Celular/efectos de los fármacos , Células Cultivadas , Cuerpo Estriado/lesiones , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/patología , Compuestos Férricos/toxicidad , Radical Hidroxilo/metabolismo , Hidroxiquinolinas/toxicidad , Inyecciones , Hierro/toxicidad , Peroxidación de Lípido , Masculino , Ratas , Ratas Wistar , SolubilidadRESUMEN
Chemical preconditioning using the mitochondrial toxin, 3-nitropropionic acid (3-NP) has been reported to induce neuroprotection against subsequent global ischemia. To investigate the underlying mechanisms, Mongolian gerbils were pretreated with either vehicle or 3-NP at the dose of 3 or 10 mg/kg, intraperitoneal, 3 days prior to a 5-min bilateral carotid artery occlusion followed by either 48 h or 7 days of blood recirculation. Neuronal damage was assessed by a cresyl violet/fuchsin acid staining. Induction of heat shock protein 72 (HSP72) and manganese superoxide dismutase (MnSOD) expression was evaluated by Western blotting. Astroglial and microglial activation was detected by immunohistochemistry (glial fibrillary acid protein) and by histochemistry (isolectin B4 staining), respectively. Present data show that the hippocampal neuronal damage induced by ischemia were of similar extent between the vehicle- and 3-NP-treated gerbils, whatever the dose tested, indicating that 3-NP did not induce tolerance to transient forebrain ischemia under our experimental conditions. The lack of difference in the post-ischemic level of HSP72 and MnSOD protein expression and in the intensity of astroglial and microglial activation represents further indirect indications of the absence of 3-NP preconditioning effect. In conclusion, although chemical preconditioning with 3-NP is a well-established phenomenon at least in vitro and in models of focal ischemia, the relevance of 3-NP as a preconditioning molecule towards global brain ischemia remains an open question.
Asunto(s)
Convulsivantes/farmacología , Hipocampo/fisiopatología , Ataque Isquémico Transitorio/fisiopatología , Precondicionamiento Isquémico , Propionatos/farmacología , Adaptación Fisiológica/fisiología , Animales , Astrocitos/química , Western Blotting , Gerbillinae , Proteína Ácida Fibrilar de la Glía/análisis , Proteínas del Choque Térmico HSP72 , Proteínas de Choque Térmico/análisis , Hipocampo/metabolismo , Ataque Isquémico Transitorio/metabolismo , Lectinas , Masculino , Microglía/química , Neuronas/química , Neuronas/enzimología , Nitrocompuestos , Superóxido Dismutasa/análisisRESUMEN
To evaluate the contribution of cellular dysfunction and neuronal loss to brain N-acetylaspartate (NAA) depletion, NAA was measured in brain tissue by HPLC and UV detection in rats subjected to cerebral injury, associated or not with cell death. When lesion was induced by intracarotid injection of microspheres, the fall in NAA was related to the degree of embolization and to the severity of brain oedema. When striatal lesion was induced by local injection of malonate, the larger the lesion volume, the higher the NAA depletion. However, reduction of brain oedema and striatal lesion by treatment with the lipophilic iron chelator dipyridyl (20 mg/kg, 1 h before and every 8 h after embolization) and the inducible nitric oxide synthase inhibitor aminoguanidine (100 mg/kg given 1 h before malonate and then every 9 h), respectively, failed to ameliorate the fall in NAA. Moreover, after systemic administration of 3-nitropropionic acid, a marked reversible fall in NAA striatal content was observed despite the lack of tissue necrosis. Overall results show that cellular dysfunction can cause higher reductions in NAA level than neuronal loss, thus making of NAA quantification a potential tool for visualizing the penumbra area in stroke patients.
Asunto(s)
Ácido Aspártico/análogos & derivados , Ácido Aspártico/análisis , Química Encefálica , Edema Encefálico/metabolismo , Isquemia Encefálica/metabolismo , 2,2'-Dipiridil/administración & dosificación , 2,2'-Dipiridil/farmacología , 2,2'-Dipiridil/uso terapéutico , Enfermedad Aguda , Animales , Biomarcadores , Edema Encefálico/etiología , Edema Encefálico/patología , Isquemia Encefálica/etiología , Isquemia Encefálica/patología , Arterias Carótidas , Muerte Celular , Cromatografía Líquida de Alta Presión , Cuerpo Estriado/efectos de los fármacos , Cuerpo Estriado/metabolismo , Cuerpo Estriado/patología , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Guanidinas/administración & dosificación , Guanidinas/farmacología , Guanidinas/uso terapéutico , Inyecciones Intraarteriales , Quelantes del Hierro/administración & dosificación , Quelantes del Hierro/farmacología , Quelantes del Hierro/uso terapéutico , Masculino , Malonatos/toxicidad , Microesferas , Mitocondrias/efectos de los fármacos , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Fármacos Neuroprotectores/administración & dosificación , Fármacos Neuroprotectores/farmacología , Fármacos Neuroprotectores/uso terapéutico , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa/antagonistas & inhibidores , Nitrocompuestos , Estrés Oxidativo , Premedicación , Propionatos/toxicidad , Ratas , Ratas Wistar , Espectrofotometría Ultravioleta , Succinato Deshidrogenasa/antagonistas & inhibidoresRESUMEN
Hypoxic preconditioning has been shown to induce neuroprotection against a subsequent damaging insult. In order to study the underlying molecular and cellular mechanisms of hypoxic preconditioning, we investigated, in gerbil hippocampus, the effects in vivo of transient exposure to hypoxia (4% O(2) for 6 min followed by either 48 h or 7 days of reoxygenation) (i) on the induction of 72 kDa heat shock protein (HSP72), heme oxygenase-1 (HO-1) and manganese superoxide dismutase (Mn SOD) as assessed by Western immunoblotting and (ii) on the astroglial and microglial activation as detected by both immunohistochemistry and Western immunoblotting for GFAP, and histochemistry for isolectin B4, respectively. Our data show that, although hypoxia and subsequent reoxygenation led to neither neuronal damage nor HSP72 induction in gerbil hippocampus, it induced a progressive and sustained expression of HO-1 and Mn SOD. As expected from the absence of neuronal death, hypoxia was not associated with microglial activation but led to a significant astrocytic activation. These findings demonstrate that transient hypoxia enhances the antioxidative enzymatic defenses of the brain, which are susceptible to increased tolerance against a subsequent damaging insult.
Asunto(s)
Hemo Oxigenasa (Desciclizante)/biosíntesis , Hipocampo/metabolismo , Hipoxia Encefálica/metabolismo , Neuroglía/metabolismo , Superóxido Dismutasa/biosíntesis , Animales , Astrocitos/citología , Astrocitos/metabolismo , Western Blotting , Gerbillinae , Proteína Ácida Fibrilar de la Glía/metabolismo , Proteínas del Choque Térmico HSP72 , Proteínas de Choque Térmico/biosíntesis , Hemo-Oxigenasa 1 , Hipocampo/citología , Inmunohistoquímica , Lectinas/metabolismo , Masculino , Neuroglía/citología , Neuronas/citología , Neuronas/metabolismo , Estrés FisiológicoRESUMEN
Iron imbalance has been implicated in oxidative injury associated with many brain diseases. The present study investigated the importance of iron location in hydroxyl radical (.OH) generation and the link between .OH production evaluated by the salicylate method and lipid peroxidation monitored by thiobarbituric acid-reactive substances assay. Brain slices were exposed to increasing doses (2, 10 and 50 microM) of Fe(III) that was complexed either to a lipophilic (8-hydroxyquinoline, HQ) or to a hydrophilic (ammoniacal citrate) ligand. Both iron complexes resulted in an increased salicylate hydroxylation and lipid peroxidation, these effects being significantly more potent in presence of Fe(III)-HQ. Salicylate hydroxylation was linearly correlated to the intensity of TBARS formation but the slope of the curve was found to be higher with Fe(III)-HQ. The present results demonstrate that 1) cell-associated reactive iron is more prone than extracellular iron to induce .OH generation, 2) the level of lipid peroxidation depending on the site of .OH production, cannot be used as an index of the level of total .OH formation, 3) the salicylate method is a convenient method to detect .OH formed intracellularly, at least in vitro.
Asunto(s)
Encéfalo/metabolismo , Radical Hidroxilo/metabolismo , Hierro/metabolismo , Animales , Encéfalo/efectos de los fármacos , Ácido Cítrico/metabolismo , Ácido Cítrico/farmacología , Compuestos Férricos/metabolismo , Compuestos Férricos/farmacología , Peroxidación de Lípido , Masculino , Oxidación-Reducción , Oxiquinolina/metabolismo , Oxiquinolina/farmacología , Ratas , Ratas Wistar , Ácido Salicílico/metabolismo , Tiobarbitúricos/metabolismoRESUMEN
BACKGROUND AND PURPOSE: To evaluate the functional prognostic value of proton magnetic resonance spectroscopy performed within the 5 days of an infarction of the middle cerebral artery territory, compared with previously demonstrated prognostic factors. METHODS: Proton magnetic resonance spectroscopy was performed on 77 consecutive non-comatosed patients during the acute stage of middle cerebral artery infarction. The functional status was determined for each patient via the Orgogozo score. Proton magnetic resonance spectroscopic data were acquired in the infarction and in contra-lateral normal tissue and the results were expressed as metabolite ratios. Correlations were evaluated between the Orgogozo score at day 1 and day 30, the age, the sex, the volume of the infarction, and the metabolic ratios. RESULTS: In a monovariate analysis, the decrease of the NAA/choline ratio was correlated with a low Orgogozo score at days 1 and 30 (P<0.05) and with a large infarction (P<0.05). A stepwise analysis showed a significant relationship between the Orgogozo score at day 30 and the Orgogozo score at day 1, the sex, the volume of infarction, and the NAA/Cho ratio within the infarction. CONCLUSIONS: Our work demonstrates that a good clinical outcome at day 30 depends on a good initial clinical score at day 1, a small volume of infarction, a small decrease of NAA/Cho, and being of the female gender.
Asunto(s)
Infarto de la Arteria Cerebral Media/diagnóstico , Espectroscopía de Resonancia Magnética , Adulto , Anciano , Femenino , Humanos , Infarto de la Arteria Cerebral Media/terapia , Masculino , Persona de Mediana Edad , Análisis Multivariante , Valor Predictivo de las Pruebas , Estudios Prospectivos , Protones , Factores de Tiempo , Resultado del TratamientoRESUMEN
Two cases in which the superior venae cavae ran an extramediastinal course are presented. This abnormality, which was discovered on catheterisation, had no haemodynamic consequences. It consisted of two superior venae cavae with little or no intermediate trunk; this explains the lack of attachment' of the superior venae cavae to the mediastinum.