RESUMEN
Long-lasting disturbances in lipid and glucose metabolism present in metabolic syndrome (MetS) lead to serious cardiovascular diseases. The study was aimed to evaluate the effect of natural antioxidant vitamin E (VitE, 100 mg/kg/day, p.o.) on basal biochemical and physiological parameters characterizing MetS and on the changed function of the heart. Furthermore, the possible potentiation of VitE effect by synthetic pyridoindole antioxidant SMe1EC2 (SMe, 15 mg/kg/day, p.o.) was also tested. MetS was induced in hereditary hypertriglyceridemic rats (HTG) by the 5 weeks administration of high-fat fructose diet (HFFD: 1 % cholesterol, 7.5 % pork lard, 10 % fructose). The heart function was tested using Langendorff preparation under constant pressure. The functional parameters of isolated heart, dysrhythmias and evoked fibrillations were evaluated in conditions of ischemia-reperfusion. The HFFD increased body weight gain and serum levels of total cholesterol, low-density lipoproteins and blood glucose. The HFFD significantly increased heart flow and force of contraction, compared to standard diet (SD). During the reperfusion, the HFFD caused the increase of the ventricular premature beats number at the expense of decreasing the duration of serious dysrhythmias (ventricular tachycardias and fibrillations). The addition of VitE, SMe or their combination to the HFFD decreased body weight gain, depressed blood pressure, improved particular biochemical parameters. The combination of VitE and SMe suppressed the occurrence of serious dysrhythmias. Our data indicate that the HFFD-related disturbances led to alterations within pathophysiology in HTG rats. The results showed that a combination of antioxidants might have the potential to amend disorders accompanying MetS.
Asunto(s)
Síndrome Metabólico , Ratas , Animales , Síndrome Metabólico/complicaciones , Síndrome Metabólico/tratamiento farmacológico , Antioxidantes/farmacología , Glucemia/metabolismo , Aumento de Peso , Dieta Alta en Grasa , FructosaRESUMEN
Metabolic syndrome (MetS) belongs to the serious health complications expanding in cardiovascular diseases, obesity, insulin resistance, and hyperglycemia. In this study, hypertriacylglycerolemic rats fed a high-fat-fructose diet (HFFD) were used as an experimental model of MetS to explore the effect of tested compounds. Effects of a new prospective pyridoindole derivative coded SMe1EC2 and the natural polyphenol rutin were tested. Endothelial nitric oxide synthase (NOS3) and nuclear factor kappa B (NF-?B) expression were assessed in the left ventricle immunohistochemically and left ventricle activity was monitored in isolated perfused rat hearts. NOS3 activity in the left ventricle decreased markedly as a result of a HFFD. NOS3 expression was upregulated by both substances. NF-?B expression was increased in the MetS group in comparison to control rats and the expression further increased in the SMe1EC2 treatment. This compound significantly improved the coronary flow in comparison to the control group during reperfusion of the heart followed after ischemia. Further, it tended to increase left ventricular systolic pressure, heart product, rate of maximal contraction and relaxation, and coronary flow during baseline assessment. Moreover, the compound SMe1EC2 decreased the sensitivity of hearts to electrically induced ventricular fibrillation. Contrary to this rutin decreased coronary flow in reperfusion. Present results suggest that despite upregulation of NOS3 by both substances tested, pyridoindole SMe1EC2 rather than rutin could be suitable in treatment strategies of cardiovascular disorders in MetS-like conditions.
Asunto(s)
Corazón/efectos de los fármacos , Indoles/uso terapéutico , Síndrome Metabólico/tratamiento farmacológico , Óxido Nítrico Sintasa de Tipo III/metabolismo , Piridinas/uso terapéutico , Rutina/uso terapéutico , Animales , Biometría , Evaluación Preclínica de Medicamentos , Fructosa/efectos adversos , Indoles/farmacología , Masculino , Síndrome Metabólico/enzimología , Síndrome Metabólico/etiología , Miocardio/metabolismo , FN-kappa B/metabolismo , Piridinas/farmacología , Ratas Wistar , Rutina/farmacologíaRESUMEN
Decreasing high fat and high carbohydrate intake, together with the administration of natural bioactive drugs, is assumed to have a protective effect in the prevention and amelioration of the metabolic syndrome (MetS). The aim of the study was to evaluate effects of diet improvement and/or a phenolic compound (rosmarinic acid; RA) administration (100 mg/kg per d) on metabolic as well as functional changes of vessels and hippocampus caused by the MetS-like conditions. The MetS-like conditions were induced by a high-fat-fructose diet (HFFD) in Prague hereditary hypertriacylglycerolaemic (HTG) rats. The effect of diet improvement and RA administration was studied using biochemical and functional measurements. Consumption of HFFD by HTG rats resulted in the development of conditions like the MetS. The fat and fructose restriction from the diet led to amelioration of basic indicators of metabolic state in rats fed HFFD and to amendment parameters of glucose tolerance test and reduction of the IL-1ß serum levels. Moreover, aortic endothelial function was improved with an impact on blood pressure. The functional measurement of electrophysiology of the hippocampus showed that long-term potentiation of neuronal transmission course deteriorated after HFFD was improved by energy restriction. Oral administration of RA had a supporting effect not only on lipid and glucose metabolism but also on the vascular endothelium. Combination of both types of therapy induced beneficial effect on glucose tolerance and lipid peroxidation. Thus, combined improvement of diet habits and treatment with natural bioactive drugs is assumed to have protective effect in prevention and amelioration of the MetS.
RESUMEN
Expression of occludin and junctional adhesion molecule A (JAM-A), transmembrane proteins of tight junctions (TJs), was analysed to characterize endothelial paracellular permeability in the heart of rats subjected to a bolus of bacterial lipopolysaccharide (LPS) at a dose of 1 mg/kg. Potential protective effects of natural carotenoids (10 mg/kg/day) produced by yeast biomass Rhodosporidium kratochvilovae on expression of occludin and JAM-A also examined in LPS-injected rats (n = 6 per group). LPS decreased expression of occludin by 40% (P < 0.01), JAM-A by 36% (P < 0.001) and increased plasma levels of malondialdehyde (MDA) and lysosomal N-acetyl-b-D-glucosaminidase (NAGA) compared to controls. Ten-day diet rich in yeast biomass containing carotenoids (torularhodin, torulene, ß-carotene) attenuated LPS-induced changes in expression of TJ proteins as observed by increased expression of occludin by 30% (P < 0.05) and JAM-A by 61% (P < 0.001) to the control values. Carotenoids also reduced oxidative stress and cellular injury indicated by decreased levels of MDA and NAGA. The results show that diet rich in yeast biomass containing natural carotenoids could protect mechanisms regulating paracellular endothelial barrier function from LPS-induced damage in the heart.
Asunto(s)
Carotenoides/farmacología , Endotelio/efectos de los fármacos , Corazón/efectos de los fármacos , Lipopolisacáridos/farmacología , Levaduras/metabolismo , Animales , Biomasa , Modelos Animales de Enfermedad , Endotelio/metabolismo , Masculino , Malondialdehído/metabolismo , Ocludina/metabolismo , Estrés Oxidativo/efectos de los fármacos , Permeabilidad/efectos de los fármacos , Ratas , Ratas Wistar , Uniones Estrechas/efectos de los fármacos , Uniones Estrechas/metabolismo , alfa-N-Acetilgalactosaminidasa/metabolismoRESUMEN
We investigated whether changes in gap junction alpha-1 protein (Cx43) expression may be associated with macrophage-induced inflammation in the heart of spontaneously hypertensive rats (SHR). To examine mutual interactions of macrophage infiltration with Cx43 expression and redistribution, we applied a bolus of bacterial lipopolysaccharide (LPS) to SHR and age-matched normotensive Wistar rats. The results demonstrated association of Cx43 downregulation with increased infiltration of cardiac CD-68 macrophages and upregulation of nuclear factor-κB (NFκB) and tumor necrosis factor-α (TNF-α) expression in the heart of SHR. LPS application to SHR caused further degradation and redistribution of Cx43 accompanied with extensively increased macrophage infiltration and NFκB and TNF-α expression. LPS administration to Wistar rats resulted in elevation of cardiac CD-68 macrophages but it did not significantly affect total Cx43 expression. Our results are suggestive of regulation of Cx43 expression with macrophages-related inflammation in the heart of SHR. The data also indicate that SHR can be more sensitive to LPS than are normotensive rats.
Asunto(s)
Conexina 43/metabolismo , Hipertensión/metabolismo , Miocardio/metabolismo , Animales , Hipertensión/patología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/fisiología , Masculino , Microscopía Electrónica de Transmisión , Miocardio/patología , Miocardio/ultraestructura , Ratas Endogámicas SHR , Ratas WistarRESUMEN
People with metabolic syndrome have higher risk of cardiovascular diseases then those without. The aim of the work was to investigate whether high fat diet administered to Prague hereditary hypertriglyceridemic (HTG) rats can induce signs of metabolic syndrome (MetS). Our results showed that HTG rats fed high fat diet (HTGch) had disturbed glucose metabolism and also lipid metabolism - increased serum triacylglycerols (TAG), total cholesterol (Ch), low-density lipoprotein-Ch (LDL-Ch), and decreased high-density lipoprotein-Ch (HDL-Ch). Their livers proved markers of developing steatosis. Moreover, HTGch had increased blood pressure, yet the vascular endothelium was not significantly damaged. All these changes were accompanied with oxidative stress and tissue damage identified as increased liver concentrations of thiobarbituric acid reactive substances (TBARS) and activity of the lysosomal enzyme N-acetyl-D-glucosaminidase (NAGA). We assume that the model used may be suitable for the study of MetS with no evidence of obesity. Prolongation of the high fat diet duration might have a major impact on all parameters tested, especially on vascular endothelial function.
Asunto(s)
Dieta Alta en Grasa/efectos adversos , Modelos Animales de Enfermedad , Hipertrigliceridemia/sangre , Hipertrigliceridemia/etiología , Síndrome Metabólico/sangre , Síndrome Metabólico/etiología , Animales , HDL-Colesterol/sangre , LDL-Colesterol/sangre , Masculino , Ratas , Ratas WistarRESUMEN
Omega-3 fatty acids (omega3FA) are known to reduce hypertriglyceridemia- and inflammation-induced vascular wall diseases. However, mechanisms of their effects are not completely clear. We examined, whether 10-day omega3FA diet can reduce bacterial lipopolysaccharide-induced changes in expression of gap junction protein connexin40 (Cx40) in the aorta of hereditary hypertriglyceridemic (hHTG) rats. After administration of a single dose of lipopolysaccharide (LPS, 1 mg/kg, i.p.) to adult hHTG rats, animals were fed with omega3FA diet (30 mg/kg/day) for 10 days. LPS decreased Cx40 expression that was associated with reduced acetylcholine-induced relaxation of aorta. Omega3FA administration to LPS rats had partial anti-inflammatory effects, associated with increased Cx40 expression and improved endothelium dependent relaxation of the aorta. Our results suggest that 10-day omega3FA diet could protect endothelium-dependent relaxation of the aorta of hHTG rats against LPS-induced damage through the modulation of endothelial Cx40 expression.
Asunto(s)
Aorta/efectos de los fármacos , Conexinas/metabolismo , Ácidos Grasos Omega-3/uso terapéutico , Hipertrigliceridemia/dietoterapia , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Aorta/metabolismo , Western Blotting , Ácidos Grasos Omega-3/farmacología , Hipertrigliceridemia/congénito , Hipertrigliceridemia/metabolismo , Lipopolisacáridos , Masculino , Ratas , Proteína alfa-5 de Unión ComunicanteRESUMEN
Connexin (Cx)-channels can represent one of targets of omega-3 fatty acids (n-3 PUFA) in protection of cardiovascular system against injury. We investigated the anti-inflammatory effect of 10-day n-3 PUFA intake (30 mg/kg/day for 10 days) on expression of Cx40 isoform in the aorta of Wistar rats injected with a single dose of lipopolysaccharide (LPS, 1 mg/kg, i.p.). LPS resulted in up-regulation of Cx40 expression in the aorta associated with reduced endothelium-dependent relaxation. LPS increased levels of inflammatory markers C-reactive protein and malondialdehyde in circulation as well as NOS activity and CD68 expression in aortic tissue indicating presence of moderate inflammation. N-3 PUFA supplementation decreased expression of both Cx40 and CD68 in aortic tissue and suppressed concentrations of C-reactive protein and malondialdehyde of endotoxemic rats. N-3 PUFA did not improve NO-dependent relaxation of aorta and NOS activity in LPS rats. The results indicate the involvement of Cx40 in development of LPS-induced endothelium-dependent functional impairment of the aorta and partial health benefits of n-3 PUFA diet associated with improved Cx40 expression.
Asunto(s)
Antiinflamatorios/farmacología , Aorta/efectos de los fármacos , Conexinas/metabolismo , Ácidos Grasos Omega-3/farmacología , Acetilglucosaminidasa/metabolismo , Animales , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Aorta/metabolismo , Aorta/fisiología , Aorta/ultraestructura , Proteína C-Reactiva/metabolismo , Células Endoteliales/efectos de los fármacos , Células Endoteliales/ultraestructura , Lipopolisacáridos , Masculino , Malondialdehído/metabolismo , Microscopía Electrónica de Transmisión , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/metabolismo , Ratas , Ratas Wistar , Vasodilatación/fisiología , Proteína alfa-5 de Unión ComunicanteRESUMEN
In the present study, we investigated the potential protective effect of selected natural substances in a rat model of heart and mesenteric ischemia-reperfusion (I/R). Experiments were performed on isolated Langendorff-perfused rat hearts, subjected to 30-min global ischemia, followed by 30-min reperfusion. Arbutin, curcumin, rosmarinic acid and extract of Mentha x villosa were applied in the concentration of 1 × 10â»5 mol/l 10 min before the onset of ischemia and during reperfusion, through the perfusion medium. Mesenteric ischemia was induced by clamping the superior mesenteric artery (SMA) for 60 min, subsequent reperfusion lasted 30 min. Production of reactive oxygen species (ROS) by SMA ex vivo was determined by luminol-enhanced chemiluminiscence (CL). The effect of the substances was tested after their incubation with tissue. Curcumin and extract of Mentha x villosa were found to be the most effective in reducing reperfusion-induced dysrhythmias--ventricular tachycardia and fibrillation. This effect was accompanied by bradycardic effect. The mesenteric I/R induced an increase in CL in vascular tissue which was dampened by substances tested. All substances tested were found to have antioxidant properties, as demonstrated by a reduction in ROS production in mesenteric vessels. This effect was confirmed in curcumin and extract of Mentha x villosa which reduced reperfusion dyshythmias.
Asunto(s)
Preparaciones de Plantas/uso terapéutico , Polifenoles/uso terapéutico , Daño por Reperfusión/tratamiento farmacológico , Animales , Antioxidantes/uso terapéutico , Arbutina/uso terapéutico , Cinamatos/uso terapéutico , Curcumina/uso terapéutico , Depsidos/uso terapéutico , Masculino , Mentha/química , Arteria Mesentérica Superior/efectos de los fármacos , Arteria Mesentérica Superior/fisiopatología , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Fitoterapia , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno , Daño por Reperfusión/fisiopatología , Taquicardia Ventricular/tratamiento farmacológico , Fibrilación Ventricular/tratamiento farmacológico , Ácido RosmarínicoRESUMEN
Local inflammation and respiratory burst of polymorphonuclear leukocytes generate reactive oxygen species (ROS). The aim of our study was to analyze the effects of peritoneal neutrophils on changes of the muscle tension of isolated aorta and compare their effects with those of different ROS. While native neutrophils did not influence muscle tension, the N-formyl-methionyl-leucyl-phenylalanine activated ones evoked a biphasic response on the KCl-precontracted aorta. The effects of activated neutrophils were in both respects similar to those evoked by xanthine/xanthine oxidase (X/XO) and differed from the effects evoked by H(2)O(2) and Fe(2)SO(4)/H(2)O(2). Using H(2)O(2) we demonstrated that the effects of ROS were dependent on the KCl induced initial tension. To exclude the effect of extensive depolarization the action of different ROS was studied also on tissues precontracted by phenylephrine. Under such condition activated neutrophils caused a marked contraction similar to that evoked by X/XO. Their effects differed however, from those of H(2)O(2) and Fe(2)SO(4)/ascorbic acid. These findings and elimination of activated neutrophil-induced contractions as well as the chemiluminiscence by superoxide dismutase suggest that the primarily activated neutrophil-released ROS was superoxide, which can be transformed to peroxynitrite, and other ROS including H(2)O(2). Reduction of all followed-up contractions caused by nordihydroguaiaretic acid, indicate that 5-lipoxygenase metabolites unselectively reduce contractions. In contrast, selective inhibition of activated neutrophil-evoked contraction by indomethacin suggests that cyclooxygenase metabolites are involved mainly in their action on vascular smooth muscle.
Asunto(s)
Aorta Torácica/fisiología , Tono Muscular/fisiología , Músculo Liso Vascular/fisiología , Neutrófilos/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Animales , Aorta Torácica/efectos de los fármacos , Cobayas , Técnicas In Vitro , Masculino , Tono Muscular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , N-Formilmetionina Leucil-Fenilalanina/farmacología , Neutrófilos/efectos de los fármacos , Fenilefrina/farmacología , Cloruro de Potasio/farmacología , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/farmacología , Vasoconstricción/efectos de los fármacos , Vasoconstricción/fisiología , Vasodilatación/efectos de los fármacos , Vasodilatación/fisiología , Xantina/metabolismo , Xantina Oxidasa/metabolismoRESUMEN
Hypertension alters expression of connexin-43 (Cx43) in cardiovascular system. The aim of the study was to investigate the effect of omega-3 polyunsaturated fatty acids (30 mg/day for 2 months) on expression of Cx43 in the aorta of 1-year-old male spontaneously hypertensive rats (SHR). Spatial distribution and expression of Cx43 in aortic wall of SHR and age-matched Lewis rats were determined by immunofluorescent method and Western blot. NO synthase (NOS) activity and endothelium-dependent relaxation of the aorta were measured as well. Immunofluorescent pattern of Cx43 was identified in endothelial and smooth muscle cells of the aorta of all experimental groups studied. However, local decrease in the number and intensity of fluorescent spots and reduced phosphorylation of Cx43 were observed in SHR in contrast to normotensive LEW. Omega-3 fatty acid diet increased Cx43 immunolabeling in endothelium and media of SHR comparing to untreated ones. Parallel, 3-fatty acids significantly elevated phosphorylation of Cx43 in the aorta of SHR (p<0.001). Despite the omega-3 fatty acids reduced blood pressure and stimulated aortic NOS activity in SHR, endothelium-dependent relaxation of the aorta did not significantly change. Results indicate that the aorta of old SHR might partially benefit from 3-PUFA supplementation due to increased Cx43 phosphorylation, NOS activity and decreased blood pressure.
Asunto(s)
Envejecimiento/metabolismo , Aorta Torácica/metabolismo , Conexina 43/metabolismo , Ácidos Grasos Omega-3/uso terapéutico , Hipertensión/metabolismo , Animales , Aorta Torácica/efectos de los fármacos , Aorta Torácica/enzimología , Presión Sanguínea/efectos de los fármacos , Western Blotting , Endotelio Vascular/efectos de los fármacos , Endotelio Vascular/enzimología , Endotelio Vascular/metabolismo , Ácidos Grasos Omega-3/administración & dosificación , Ácidos Grasos Omega-3/farmacología , Técnica del Anticuerpo Fluorescente , Hipertensión/tratamiento farmacológico , Hipertensión/fisiopatología , Masculino , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/enzimología , Músculo Liso Vascular/metabolismo , Óxido Nítrico Sintasa/metabolismo , Fosforilación , Ratas , Ratas Endogámicas Lew , Ratas Endogámicas SHRAsunto(s)
Antagonistas de los Receptores Histamínicos H1/farmacología , Mesenterio , Feniramina/farmacología , Daño por Reperfusión , Animales , Bromofeniramina/farmacología , Clorfeniramina/farmacología , Masculino , Mesenterio/efectos de los fármacos , Mesenterio/patología , Neutrófilos/inmunología , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/patología , Daño por Reperfusión/prevención & controlRESUMEN
Diabetes mellitus is associated with many complications including retinopathy, nephropathy, neuropathy and angiopathy. Increased cardiovascular risk is accompanied with diabetes-induced endothelial dysfunction. Pharmacological agents with endothelium-protective effects may decrease cardiovascular complications. In present study sulodexide (glycosaminoglycans composed from heparin-like and dermatan fractions) was chosen to evaluate its protective properties on endothelial dysfunction in diabetes. Effect of sulodexide treatment (SLX, 100 UI/kg/day, i.p.) in 5 and 10 weeks lasting streptozotocin-induced diabetes (30 mg/kg/day, i.p. administered for three consecutive days) was investigated. Animals were divided into four groups: control (injected with saline solution), control-treated with sulodexide (SLX), diabetic (DM) and diabetic-treated with sulodexide (DM+SLX). The pre-prandial and postprandial plasma glucose levels, number of circulating endothelial cells (EC) and acetylcholine-induced relaxation of isolated aorta and mesenteric artery were evaluated. Streptozotocin elicited hyperglycemia irrespective of SLX treatment. Streptozotocin-induced diabetes enhanced the number of circulating endothelial cells compared to controls. SLX treatment decreased the number of EC in 10-week diabetes. Acetylcholine-induced relaxation of mesenteric arteries was significantly impaired in 5 and 10-week diabetes. SLX administration improved relaxation to acetylcholine in 5 and 10-week diabetes. Diabetes impaired acetylcholine-induced relaxation of rat aorta irrespective of SLX treatment. Our results demonstrate that SLX treatment lowers the number of circulating endothelial cells and improves endothelium-dependent relaxation in small arteries. These findings suggest endothelium-protective effect of sulodexide in streptozotocin-induced diabetes.
Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Angiopatías Diabéticas/prevención & control , Endotelio Vascular/efectos de los fármacos , Glicosaminoglicanos/farmacología , Hipoglucemiantes/farmacología , Vasodilatación/efectos de los fármacos , Acetilcolina/farmacología , Animales , Glucemia/efectos de los fármacos , Diabetes Mellitus Experimental/fisiopatología , Angiopatías Diabéticas/fisiopatología , Relación Dosis-Respuesta a Droga , Endotelio Vascular/fisiopatología , Masculino , Ratas , Ratas Wistar , Factores de Tiempo , Vasodilatadores/farmacologíaAsunto(s)
Antagonistas de los Receptores Histamínicos H1/metabolismo , Isquemia , Mesenterio , Daño por Reperfusión , Animales , Benzotiepinas/farmacología , Loratadina/farmacología , Masculino , Mesenterio/irrigación sanguínea , Mesenterio/efectos de los fármacos , Mesenterio/patología , Modelos Biológicos , Ratas , Ratas Wistar , Daño por Reperfusión/prevención & controlRESUMEN
This study examined the effects of the pyridoindole compound stobadine on intestinal and vascular injury following mesenteric ischemia/reperfusion (I/R) in rats. Ischemia was induced by occlusion of the superior mesenteric artery (SMA) for 60 min, followed by 30 min reperfusion. To characterize gut impairment, some parameters of intestinal damage and biochemical variables, such as GSH content, activity of a lysosomal enzyme N-acetyl-beta-D-glucuronidase and activity of gamma-glutamyl transpeptidase, were determined. Vascular I/R-induced damage was evaluated as changes in acetylcholine evoked relaxation of mesenteric artery rings under isometric conditions. A method of amplified chemiluminescence (CL) was used to detect production of reactive oxygen species (ROS). Following I/R, pronounced intestinal injury of various intensities was observed, with maximal changes occurring in the terminal ileum. The effect of I/R was expressed mainly as increased vascular permeability, with protein leakage and subsequent hemorrhagic injury of the intestine as well as impaired endothelium-dependent SMA relaxation. Vessel dysfunction was manifested by a decrease of the maximal relaxation response to acetylcholine. An increase of CL, indicative of increased ROS production, was observed in both intestinal and vascular tissue. A novel antioxidant, stobadine, was found to reduce the increased vascular permeability and the extent of small intestine injury caused by I/R, to improve biochemical alterations accompanying I/R, to protect endothelial-dependent relaxation of mesenteric arteries, and to attenuate the CL response. The observed beneficial effect of stobadine indicates its possible application in the preventive and/or therapeutic approach to I/R-induced pathologies.
Asunto(s)
Antioxidantes/farmacología , Carbolinas/farmacología , Intestino Delgado/efectos de los fármacos , Arteria Mesentérica Superior/efectos de los fármacos , Daño por Reperfusión/tratamiento farmacológico , Acetilcolina , Animales , Permeabilidad Capilar , Endotelio Vascular , Glucuronidasa/efectos de los fármacos , Glucuronidasa/metabolismo , Glutatión/efectos de los fármacos , Glutatión/metabolismo , Íleon/efectos de los fármacos , Íleon/fisiopatología , Intestino Delgado/patología , Luminiscencia , Arteria Mesentérica Superior/fisiopatología , Transporte de Proteínas , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/fisiopatología , gamma-Glutamiltransferasa/efectos de los fármacos , gamma-Glutamiltransferasa/metabolismoRESUMEN
In this study we investigated functional changes in the femoral artery and ultrastructural alterations in mesenteric vessels and capillaries in the rat model of multiple low dose streptozotocin (STZ)-induced diabetes. Participation of oxidative stress in this model of diabetes was established by studying the effect of the pyridoindole antioxidant stobadine (STB) on diabetes-induced impairment. Experimental diabetes was induced by i.v. bolus of STZ (20 mg/kg) given for three consecutive days to male rats. At the 12(th) week following STZ administration, the animals revealed typical signs of diabetes, such as polyphagia, polydypsia and polyuria. There was no weight gain in the diabetic groups throughout the experiment. No exitus occurred in any group. Diabetes was characterised with high levels of plasma glucose, no significant changes in lipid metabolism, decreased serum levels of glutathione, increased serum levels of the lysosomal enzyme N-acetyl-beta-D-glucosaminidase (NAGA), injured endothelial relaxant capacity of the femoral artery and alterations in ultrastructure of mesenteric arteries and capillaries. Antioxidant STB in the dose of 25 mg/kg body weight i.p. (5 times per week) did not influence glucose levels, however, it mitigated biochemical, functional and ultrastructural changes induced by diabetes, suggesting a role of reactive oxygen species in diabetes-induced tissue damage.
Asunto(s)
Vasos Sanguíneos/efectos de los fármacos , Diabetes Mellitus Experimental/patología , Diabetes Mellitus Experimental/fisiopatología , Animales , Antioxidantes/farmacología , Vasos Sanguíneos/patología , Vasos Sanguíneos/fisiopatología , Carbolinas/farmacología , Arteria Femoral/efectos de los fármacos , Arteria Femoral/fisiopatología , Técnicas In Vitro , Masculino , Arterias Mesentéricas/efectos de los fármacos , Arterias Mesentéricas/patología , Microscopía Electrónica , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Estreptozocina/administración & dosificación , Estreptozocina/toxicidad , Vasodilatación/efectos de los fármacosRESUMEN
(2-benzyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indole-8-yl)-acetic acid (compound 1), a novel aldose reductase inhibitor, was assayed for efficacy and selectivity to inhibit rat lens aldose reductase under in vitro conditions by using enzyme preparations obtained from diabetic animals. The inhibitory efficiency was characterized by IC(50) in micromolar region. Enzyme kinetics analysis revealed uncompetitive type of inhibition, both in relation to the D,L-glyceraldehyde substrate and to the NADPH cofactor. In testing for selectivity, comparisons to rat kidney aldehyde reductase, an enzyme with the highest homology to aldose reductase, was used. The inhibition selectivity of the compound tested was characterized by selectivity factor around 20 and was even slightly improved under conditions of prolonged experimental diabetes. These findings were identical with those in the control rats. To conclude, the inhibitory mode, efficacy and selectivity of compound 1, a novel aldose reductase inhibitor, was preserved even under the conditions of prolonged STZ-induced experimental diabetes of rats.
Asunto(s)
Acetatos/farmacología , Aldehído Reductasa/antagonistas & inhibidores , Diabetes Mellitus Experimental/inducido químicamente , Diabetes Mellitus Experimental/enzimología , Indoles/farmacología , Riñón/enzimología , Cristalino/enzimología , Animales , Femenino , Masculino , Ratas , Ratas Wistar , EstreptozocinaRESUMEN
A rat model of transient occlusion of the superior mesenteric artery was used to study the intestinal and vascular injury induced by ischaemia/ reperfusion (I/R). A pronounced intestinal injury was observed, ranging from hyperaemia to severe haemorrhagic necrosis and bleeding. The length of the damaged intestinal segments reached 58.6% of the small intestine with a decrease of the wet weight in the I/R group. Sham operation resulted in 100% survival, I/R decreased survival to 40% after 24 h. Following I/R a significant increase of vascular permeability was observed in the small intestine. Gamma-glutamyl transpeptidase activity decreased aborally in sham operated rats and I/R reduced it further in all parts of the small intestine. I/R resulted in damaged endothelium-dependent relaxation of mesenteric artery rings. This was manifested by decreased maximal responses of arterial preparations to acetylcholine as well as decreased pD2 values. The results confirmed and specified the presumed effect of I/R on the small intestine and on vascular functions.
Asunto(s)
Intestinos/irrigación sanguínea , Isquemia/fisiopatología , Daño por Reperfusión/fisiopatología , Análisis de Varianza , Animales , Modelos Animales de Enfermedad , Masculino , Arterias Mesentéricas , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/cirugíaRESUMEN
Metabolism, monitored via in situ catalytic enzyme histochemistry and fine structure, was studied in the myocardium of chronic diabetic male Wistar rats administered L-arginine (12.8 mg/100 g/day) for 24 weeks. Diabetes was induced with a single i.v. injection of 55 mg/kg streptozotocin. After 6 months, the tissue of the left ventricle was processed for electron microscope examination and transmural tissue blocks were frozen for enzyme histochemistry. In diabetic myocardium, heterogeneous ischemia-like subcellular alterations of cardiomyocytes and capillaries were observed, together with interstitial fibrosis. This structural remodeling was accompanied by significantly decreased activity of endothelial nitric oxide synthase (NOS) and heterogeneously decreased activities of glycogen phosphorylase (GlPh), hydroxybutyrate dehydrogenase (HBDH) and adenosine triphophatases (ATPases) throughout the myocardium. In arginine-treated diabetic rats, there was evidence of protected structural integrity of endothelial cells and attenuated structural disturbances of cardiomyocytes. This was associated with the markedly preserved histochemical activities of all detected enzymes in comparison with nontreated diabetic rats (NOS 98.7 +/- 10.5% vs. 35.4 +/- 4.1%; ATPases 82.7 +/- 9.1% vs. 69.3 +/- 5.2%; GlPh 65.2 +/- 8.3% vs. 45.5 +/- 3.8%; HBDH 68.9 +/- 8.5% vs. 44.1 +/- 6.7% of control values). The results indicate that long-term supplementation of L-arginine may account for the reduction of diabetes-induced myocardial structural remodeling.
Asunto(s)
Arginina/uso terapéutico , Diabetes Mellitus Experimental/metabolismo , Miocardio/metabolismo , Remodelación Ventricular/efectos de los fármacos , Animales , Diabetes Mellitus Experimental/tratamiento farmacológico , Masculino , Miocardio/enzimología , Miocardio/ultraestructura , Óxido Nítrico Sintasa/metabolismo , Ratas , Ratas WistarRESUMEN
We studied the influence of hyperglycemia lasting 1, 4, 6 and 8 months on the reactivity and ultrastructure of the aorta in Wistar rats. Moreover, the effect of the pyridoindole antioxidant stobadine ((-)-cis-2,8-dimethyl-2,3,4,4a,5,9b-hexahydro-1H-pyrido[4,3-b]indole) on the changes induced by the 8-month hyperglycemia were studied. Hyperglycemia was induced by streptozotocin (STZ, 55 mg/kg i.v.). In the functional study, responses to KCl, acetylcholine (ACh), noradrenaline (NA) and hydrogen peroxide were evaluated under isometric conditions. The first changes in aortic reactivity started after 1 month of hyperglycemia and were exhibited by significantly increased NA-induced contractions. Relaxant responses to acetylcholine were decreased, although not significantly. Prolongation of hyperglycemia to 4, 6 and 8 months did not cause any additional significant changes in responsiveness to NA. Decreased ACh-induced relaxation and increased contractile responses to H2O2 were observed in month 4. The functional responses were not substantially deteriorated by prolongation of hyperglycemia to 6 and 8 months. Ultrastructural examination of the diabetic aorta showed disturbances in normal tissue organization. An 8-month supplementation of stobadine in diabetic rats resulted in the protection of aortic function as well as its ultrastructure. These results suggest that abnormalities occurring in the aorta of diabetic rats might result from the damaging effects of oxygen free radicals.