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BACKGROUND: The effects of hypertension and exercise training (T) on the sequential interplay between renin-angiotensin system (RAS), autonomic control and heart remodeling during the development of hypertension in spontaneously hypertensive rats (SHR), was evaluated.MethodsâandâResults:Time course changes of these parameters were recorded in 4-week-old SHR submitted to a T or sedentary (S) protocol. Wistar Kyoto rats served as controls. Hemodynamic recordings were obtained in conscious rats at experimental weeks 0, 1, 2, 4, and 8. The left ventricle (LV) was collected to evaluate RAS gene and protein expression, cardiomyocytes' hypertrophy and collagen accumulation. Pre-hypertensive SHR exhibited augmented AT1R gene expression; at 5 weeks, they presented with elevated pressure, increased LV angiotensinogen and ACE mRNA expression, followed by sympathoexcitation (from the 8thweek onwards). Marked AT1R protein content, myocytes's hypertrophy, collagen deposition and increased pressure variability were observed in 12-week-old sedentary SHR. In addition to attenuating all these effects, T activated Mas receptor expression augmented parasympathetic modulation of the heart, and delayed the onset and reduced the magnitude, but did not block the development of genetic hypertension. CONCLUSIONS: The close temporal relationship between changes in the LV ACE-Ang II-AT1R axis, autonomic control and cardiac remodeling at both the establishment of hypertension and during exercise training reveals the essential role played by the AT1R pathway in driving cardiac remodeling and autonomic modulation during the transition from the pre- to hypertensive phase.
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Terapia por Ejercicio , Corazón/inervación , Hipertensión/prevención & control , Prehipertensión/terapia , Receptor de Angiotensina Tipo 1/metabolismo , Sistema Nervioso Simpático/fisiopatología , Función Ventricular Izquierda , Remodelación Ventricular , Animales , Presión Sanguínea , Modelos Animales de Enfermedad , Regulación hacia Abajo , Hipertensión/genética , Hipertensión/metabolismo , Hipertensión/fisiopatología , Prehipertensión/genética , Prehipertensión/metabolismo , Prehipertensión/fisiopatología , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Receptor de Angiotensina Tipo 1/genética , Transducción de Señal , Factores de TiempoRESUMEN
INTRODUCTION: In spite of the great relevance of abdominal aortic aneurysm, its etiopathogenesis is not fully understood. The biomechanical and histological study of the aortic wall may contribute to this elucidation. METHODS: Seventy-five male Wistar rats were divided into 4 groups: control (CG), smoker (SG), diabetic (DG), and diabetic + smoker (DSG). The SG and DSG rats were exposed to cigarette smoke for 30 min/day, 5 days a week. Diabetes was induced by the intravenous injection of streptozotocin. After 16 weeks, the abdominal aorta was collected for biomechanical, histological, and matrix metalloproteinase 2 (MMP-2) activity analyses. RESULTS: The valid biomechanical tests of 52 specimens were analyzed: 11 in the CG, 10 in the DG, 16 in the SG, and 15 in the DSG. The biomechanical analysis of the fragments showed no differences between the control, DG, SG, and DSG. Collagen deposition also did not present a significant difference between the studied groups. The total count of elastic fibers was higher in diabetic rats (DG and DSG) than in the SG. The inflammatory response observed in all experimental groups was significantly more intense than in the CG. Compared to the DSG, MMP-2 activity showed a significant decrease in the DG. CONCLUSIONS: Resistance and elasticity did not present a difference between the CG and the DG, SG, and DSG. Compared to the CG, the total count of elastic fibers, fragmentation of the elastic lamina, pericellular matrix deposition, and cell loss/substitution in the tunica media showed significant alterations in the aortic walls of the DG, SG, and DSG. MMP-2 activity was lower in the DG aorta than in the DSG aorta.
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Aorta Abdominal/patología , Aneurisma de la Aorta Abdominal/etiología , Diabetes Mellitus Experimental/complicaciones , Humo/efectos adversos , Productos de Tabaco/efectos adversos , Animales , Aorta Abdominal/metabolismo , Aorta Abdominal/fisiopatología , Aneurisma de la Aorta Abdominal/metabolismo , Aneurisma de la Aorta Abdominal/patología , Aneurisma de la Aorta Abdominal/fisiopatología , Fenómenos Biomecánicos , Colágeno/metabolismo , Progresión de la Enfermedad , Tejido Elástico/patología , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Ratas Wistar , Flujo Sanguíneo Regional , Factores de Riesgo , Estrés Mecánico , Factores de TiempoRESUMEN
We tested the hypothesis that an increase in the anti-inflammatory cholinergic pathway, when induced by pyridostigmine (PY), may modulate subtypes of lymphocytes (CD4+, CD8+, FOXP3+) and macrophages (M1/M2) soon after myocardial infarction (MI) in rats. Wistar rats, randomly allocated to receive PY (40 mg·kg(-1)·day(-1)) in drinking water or to stay without treatment, were followed for 4 days and then were subjected to ligation of the left coronary artery. The groups-denominated as the pyridostigmine-treated infarcted (IP) and infarcted control (I) groups-were submitted to euthanasia 3 days after MI; the heart was removed for immunohistochemistry, and the peripheral blood and spleen were collected for flow cytometry analysis. Noninfarcted and untreated rats were used as controls (C Group). Echocardiographic measurements were registered on the second day after MI, and heart rate variability was measured on the third day after MI. The infarcted groups had similar MI areas, degrees of systolic dysfunction, blood pressures, and heart rates. Compared with the I Group, the IP Group showed a significant higher parasympathetic modulation and a lower sympathetic modulation, which were associated with a small, but significant, increase in diastolic function. The IP Group showed a significant increase in M2 macrophages and FOXP3(+)cells in the infarcted and peri-infarcted areas, a significantly higher frequency of circulating Treg cells (CD4(+)CD25(+)FOXP3(+)), and a less extreme decrease in conventional T cells (CD25(+)FOXP3(-)) compared with the I Group. Therefore, increasing cholinergic modulation with PY induces greater anti-inflammatory cell recruitment soon after MY in rats.
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Antiinflamatorios/farmacología , Linfocitos T CD8-positivos/efectos de los fármacos , Quimiotaxis de Leucocito/efectos de los fármacos , Neuronas Colinérgicas/efectos de los fármacos , Inhibidores de la Colinesterasa/farmacología , Macrófagos/efectos de los fármacos , Infarto del Miocardio/tratamiento farmacológico , Miocardio/inmunología , Bromuro de Piridostigmina/farmacología , Linfocitos T Reguladores/efectos de los fármacos , Animales , Biomarcadores/metabolismo , Presión Sanguínea/efectos de los fármacos , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Neuronas Colinérgicas/metabolismo , Neuronas Colinérgicas/patología , Modelos Animales de Enfermedad , Factores de Transcripción Forkhead/metabolismo , Frecuencia Cardíaca/efectos de los fármacos , Subunidad alfa del Receptor de Interleucina-2/metabolismo , Activación de Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Infarto del Miocardio/inmunología , Infarto del Miocardio/metabolismo , Infarto del Miocardio/patología , Infarto del Miocardio/fisiopatología , Miocardio/metabolismo , Miocardio/patología , Fenotipo , Ratas Endogámicas WKY , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/metabolismo , Linfocitos T Reguladores/inmunología , Linfocitos T Reguladores/metabolismo , Factores de Tiempo , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
BACKGROUND: The estrogen deficiency, abnormal lipid profile, weight gain and a sedentary lifestyle are factors associated with the increased prevalence of cardiovascular disease in menopausal women. However, physical exercise practice reduces some of these risk factors. Moreover, it has been shown that exercise has an impact on inflammation, in sympathetic activity and improves endothelial function. AIMS: The present study aims to evaluate the effects of moderate aerobic training on biochemical, morphological and physiological parameters in LDL Knockout mice with estrogen deprivation, evaluating the components of the ascending aortic wall. METHODS: The animals were randomly divided into six groups (n=5): sedentary control (SC), sedentary control ovariectomized (SCO), trained control ovariectomized (TCO), LDL-Knockout sedentary (KS), LDL-Knockout sedentary ovariectomized (KOS) and LDL-Knockout trained ovariectomized (KOT). The trained groups underwent a protocol of moderate training for 4 weeks on a treadmill with speed and progressive load. After training, blood samples were collected for biochemical assessments and the aorta was removed for dissection and histological morphometry study. In addition, the expression of angiotensin-converting enzyme (ACE) and angiotensin II proteins were examined by immunohistochemistry in all groups of animals. RESULTS: Changes of expressions of ACE and angiotensin II were found when the group was subjected to exercise. The concentrations of cholesterol and triglycerides were lower in the groups of animals with estrogen deprivation and dyslipidemia. In animals that performed exercises we found significant increase (p<0.05) in Vv[lam]; decrease in Vv[col] and CWT, and a tendency for decrease both in TS and IMT when compared to the SC groups. The histological morphometry findings showed consistency in the results of the aorta study when the ovariectomized group underwent the exercise protocol. CONCLUSION: We conclude that physical training contributed to reducing vessel rigidity and to improvements in vascular compliance, with the increase in volume density of elastic lamellae in the estrogen-deprived groups who had normal cholesterol levels.
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Aorta/fisiopatología , Angiotensina II/metabolismo , Animales , Aorta/metabolismo , Presión Sanguínea , Peso Corporal , Colesterol/sangre , Dislipidemias/metabolismo , Dislipidemias/patología , Módulo de Elasticidad , Estrógenos/sangre , Femenino , Frecuencia Cardíaca , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Modelos Animales , Ovariectomía , Peptidil-Dipeptidasa A/metabolismo , Condicionamiento Físico Animal , Resistencia a la Tracción , Triglicéridos/sangreRESUMEN
In the present study we evaluated the effects of short-term pyridostigmine bromide (0.14 mg/mL) treatment started early after myocardial infarction (MI) on left ventricular (LV) and autonomic functions in rats. Male Wistar rats were divided into control, pyridostigmine, infarcted and infarcted + pyridostigmine-treated groups. Pyridostigmine was administered in the drinking water, starting immediately after MI or sham operation, for 11 days. Left ventricular function was evaluated indirectly by echocardiography and directly by LV catheterization. Cardiovascular autonomic control was evaluated by baroreflex sensitivity (BRS), heart rate variability (HRV) and pharmacological blockade. All evaluations started after 7 days pyridostigmine treatment and were finalized after 11 days treatment. Pyridostigmine prevented the impairment of +dP/dT and reduced the MI area in infarcted + pyridostigmine compared with infarcted rats (7 ± 3% vs 17 ± 4%, respectively). Mean blood pressure was restored in infarcted + pyridostigmine compared with infarcted rats (103 ± 3 vs 94 ± 3 mmHg, respectively). In addition, compared with the infarcted group, pyridostigmine improved BRS, as evaluated by tachycardic (1.6 ± 0.2 vs 2.5 ± 0.2 b.p.m./mmHg, respectively) and bradycardic (-0.42 ± 0.01 vs -1.9 ± 0.1 b.p.m./mmHg) responses, and reduced the low frequency/high frequency ratio of HRV (0.81 ± 0.11 vs 0.24 ± 0.14, respectively). These improvements are probably associated with increased vagal tone and reduced sympathetic tone in infarcted + pyridostigmine compared with infarcted rats. In conclusion, the data suggest that short-term pyridostigmine treatment started early after MI can improve BRS, HRV and parasympathetic and sympathetic tone in experimental rats. These data may have potential clinical implications because autonomic markers have prognostic significance after MI.
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Colinérgicos/farmacología , Infarto del Miocardio/tratamiento farmacológico , Bromuro de Piridostigmina/farmacología , Animales , Barorreflejo/efectos de los fármacos , Presión Sanguínea/efectos de los fármacos , Ecocardiografía/métodos , Frecuencia Cardíaca/efectos de los fármacos , Ventrículos Cardíacos/efectos de los fármacos , Masculino , Ratas , Ratas Wistar , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
BACKGROUND: NOX4 activation has been implicated to have vasoprotective and blood pressure (BP)-lowering effects. Molecular mechanisms underlying this are unclear, but NOX4-induced regulation of the redox-sensitive Ca 2+ channel TRPM2 and effects on endothelial nitric oxide synthase (eNOS)-nitric oxide signalling may be important. METHOD: Wild-type and LinA3, renin-expressing hypertensive mice, were crossed with NOX4 knockout mice. Vascular function was measured by myography. Generation of superoxide (O 2- ) and hydrogen peroxide (H 2 O 2 ) were assessed by lucigenin and amplex red, respectively, and Ca 2+ influx by Cal-520 fluorescence in rat aortic endothelial cells (RAEC). RESULTS: BP was increased in NOX4KO, LinA3 and LinA3/NOX4KO mice. This was associated with endothelial dysfunction and vascular remodelling, with exaggerated effects in NOX4KO groups. The TRPM2 activator, ADPR, improved vascular relaxation in LinA3/NOX4KO mice, an effect recapitulated by H 2 O 2 . Inhibition of PARP and TRPM2 with olaparib and 2-APB, respectively, recapitulated endothelial dysfunction in NOX4KO. In endothelial cells, Ang II increased H 2 O 2 generation and Ca 2+ influx, effects reduced by TRPM2 siRNA, TRPM2 inhibitors (8-br-cADPR, 2-APB), olaparib and GKT137831 (NOX4 inhibitor). Ang II-induced eNOS activation was blocked by NOX4 and TRPM2 siRNA, GKT137831, PEG-catalase and 8-br-cADPR. CONCLUSION: Our findings indicate that NOX4-induced H 2 O 2 production activates PARP/TRPM2, Ca 2+ influx, eNOS activation and nitric oxide release in endothelial cells. NOX4 deficiency impairs Ca 2+ homeostasis leading to endothelial dysfunction, an effect exacerbated in hypertension. We define a novel pathway linking endothelial NOX4/H 2 O 2 to eNOS/nitric oxide through PARP/TRPM2/Ca 2+ . This vasoprotective pathway is perturbed when NOX4 is downregulated and may have significance in conditions associated with endothelial dysfunction, including hypertension.
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Hipertensión , Canales Catiónicos TRPM , Animales , Ratones , Ratas , Calcio/metabolismo , Células Endoteliales/metabolismo , Peróxido de Hidrógeno/farmacología , Hipertensión/metabolismo , Óxido Nítrico/metabolismo , Inhibidores de Poli(ADP-Ribosa) Polimerasas , Canales Catiónicos TRPM/genética , Canales Catiónicos TRPM/metabolismoRESUMEN
PURPOSE: To evaluate the effects of the experimental subcutaneous Walker-256 tumor and L-glutamine supplementation, an antioxidant, on the glomerular morphology of rats. METHODS: Twenty Wistar rats were distributed into four groups (n = 5): control (C); control treated with 2% L-glutamine (CG); rats with Walker-256 tumor (WT); and rats with Walker-256 tumor treated with 2% L-glutamine (WTG). Renal histological samples were submitted to periodic acid-Schiff and Masson's Trichrome staining to analyze glomerular density, morphometry of glomerular components and glomerulosclerosis; and to immunohistochemistry for fibroblast growth factor-2 (FGF-2). RESULTS: WT showed 50% reduction in body mass gain and cachexia index > 10%, while WTG demonstrated reduction in cachexia (p < 0.05). WT revealed reduction of glomerular density, increase in the glomerular tuft area, mesangial area, matrix in the glomerular tuft, decrease in the urinary space and synechia, and consequently higher glomerulosclerosis (p < 0.05). L-glutamine supplementation in the WTG improved glomerular density, and reduced glomerular tuft area, urinary space, mesangial area, and glomerulosclerosis compared to WT(p < 0.05). WT showed higher collagen area and FGF-2 expression compared to C (p < 0.05). WTG presented lower collagen fibers and FGF-2 expression compared to WT (p < 0.05). CONCLUSIONS: L-glutamine supplementation reduced cachexia and was beneficial for glomerular morphology of the rats, as well as it reduced kidney damage and improved the remaining glomeruli morphology.
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Glutamina , Neoplasias , Ratas , Animales , Ratas Wistar , Glutamina/farmacología , Caquexia/metabolismo , Caquexia/patología , Factor 2 de Crecimiento de Fibroblastos , Suplementos Dietéticos , ColágenoRESUMEN
BACKGROUND: Exercise training (ET) has been used as a nonpharmacological strategy for treatment of diabetes and myocardial infarction (MI) separately. We evaluated the effects ET on functional and molecular left ventricular (LV) parameters as well as on autonomic function and mortality in diabetics after MI. METHODS AND RESULTS: Male Wistar rats were divided into control (C), sedentary-diabetic infarcted (SDI), and trained-diabetic infarcted (TDI) groups. MI was induced after 15 days of streptozotocin-diabetes induction. Seven days after MI, the trained group underwent ET protocol (90 days, 50-70% maximal oxygen consumption-VO(2)max). LV function was evaluated noninvasively and invasively; baroreflex sensitivity, pulse interval variability, cardiac output, tissue blood flows, VEGF mRNA and protein, HIF1-α mRNA, and Ca(2+) handling proteins were measured. MI area was reduced in TDI (21 ± 4%) compared with SDI (38 ± 4%). ET induced improvement in cardiac function, hemodynamics, and tissue blood flows. These changes were probable consequences of a better expression of Ca(2+) handling proteins, increased VEGF mRNA and protein expression as well as improvement in autonomic function, that resulted in reduction of mortality in TDI (33%) compared with SDI (68%) animals. CONCLUSIONS: ET reduced cardiac and peripheral dysfunction and preserved autonomic control in diabetic infarcted rats. Consequently, these changes resulted in improved VO(2)max and survival after MI.
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Sistema Nervioso Autónomo , Circulación Coronaria , Corazón/fisiopatología , Infarto del Miocardio/patología , Condicionamiento Físico Animal , Análisis de Varianza , Animales , Calcio/metabolismo , Gasto Cardíaco , Hemodinámica , Masculino , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/prevención & control , Consumo de Oxígeno , Ratas , Ratas Wistar , Factores de Tiempo , Ultrasonografía , Factor A de Crecimiento Endotelial VascularRESUMEN
AIMS: To test the effects of early exercise training (ET) on left ventricular (LV) and autonomic functions, haemodynamics, tissues blood flows (BFs), maximal oxygen consumption (VO(2) max), and mortality after myocardial infarction (MI) in rats. METHODS AND RESULTS: Male Wistar rats were divided into: control (C), sedentary-infarcted (SI), and trained-infarcted (TI). One week after MI, TI group underwent an ET protocol (90 days, 50-70% VO(2) max). Left ventricular function was evaluated non-invasively and invasively. Baroreflex sensitivity, heart rate variability, and pulse interval were measured. Cardiac output (CO) and regional BFs were determined using coloured microspheres. Infarcted area was reduced in TI (19 ± 6%) compared with SI (34 ± 5%) after ET. Exercise training improved the LV and autonomic functions, the CO and regional BF changes induced by MI, as well as increased SERCA2 expression and mRNA vascular endothelial growth factor levels. These changes brought about by ET resulted in mortality rate reduction in the TI (13%) group compared with the SI (54%) group. CONCLUSION: Early aerobic ET reduced cardiac and peripheral dysfunctions and preserved cardiovascular autonomic control after MI in trained rats. Consequently, these ET-induced changes resulted in improved functional capacity and survival after MI.
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Enfermedades del Sistema Nervioso Autónomo/fisiopatología , Infarto del Miocardio/fisiopatología , Condicionamiento Físico Animal , Animales , Barorreflejo/fisiología , Peso Corporal , Gasto Cardíaco/fisiología , Ecocardiografía , Frecuencia Cardíaca/fisiología , Estimación de Kaplan-Meier , Masculino , Microesferas , Infarto del Miocardio/mortalidad , Consumo de Oxígeno/fisiología , Ratas , Ratas Wistar , Flujo Sanguíneo Regional/fisiología , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Disfunción Ventricular Izquierda/fisiopatologíaRESUMEN
Angiotensin-converting enzyme I (ACE) is a key part of the renin-angiotensin system. Its main function is to regulate blood pressure and the balance of salts in the body. Somatic ACE has two domains, N-C-, each of which has a catalytic site that exhibits 60%sequence identity. The N-domain has a specific action in the hydrolysis of beta-amyloid bodies and angiotensin (1-7), which activates the MAS receptor and triggers anti-thrombotic and anti-inflammatory actions. Our goal was to obtain the catalytic site Ala361 to Gly468 of the N domain region, csACEN, without needing purification by chromatography. We employed a method that uses an Elastin-like Polypeptide (ELP) and Intein sequences linked to the peptide of interest. The more differential for obtaining the pure peptide was the cultivation temperatures in the synthesis of ELPcsACEN at 37 °C, with a significant increase in expression. In the purification by ELP precipitation, we recorded the highest efficiency in the concentrations of 0.57 M and 0.8 M of ammonium sulfate buffer. Intein autocleavage study allows removal of the ELP sequence at acidic pH, with the buffers MES and Tris-HCl The present study defined the best conditions for obtaining pure csACEN that the literature has not yet described for peptides. Obtaining pure csACEN aims at future studies for therapeutic use in hypertension, Alzheimer's, and oncology.
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Elastina , Inteínas , Angiotensinas , Dominio Catalítico , Elastina/química , Elastina/metabolismo , Péptidos/químicaRESUMEN
This study aimed to analyze the effects of the quercetin (100 mg/kg), 1% glutamine and 1% α-tocopherol antioxidants in the myocardium of rats with streptozotocin-induced diabetes mellitus. Twenty male rats were subdivided into four groups (n = 5): N (normoglycemic); D (diabetic); NT (normoglycemic treated with antioxidants); and DT (diabetic treated with antioxidants) treated for 60 days. Clinical parameters, oxidative stress markers, inflammatory cytokines, myocardial collagen fibers and immunoexpression of superoxide dismutase 1 (SOD-1), glutathione peroxidase-1 (GPx-1), interleukin-1ß (IL-1-ß), transforming growth factor-beta (TGF-ß), and fibroblast growth factor-2 (FGF-2) were evaluated. Results showed reduced body weight, hyperphagia, polydipsia and hyperglycemic state in groups D and DT. The levels of glutathione (GSH) were higher in NT and DT compared to N (p < 0.01) and D (p < 0.001) groups, respectively. Greater GSH levels were found in DT when compared to N animals (p < 0.001). In DT, there was an increase in IL-10 in relation to N, D and NT (p < 0.05), while GPx-1 expression was similar to N and lower compared to D (p < 0.001). TGF-ß expression in DT was greater than N (p < 0.001) group, whereas FGF-2 in DT was higher than in the other groups (p < 0.001). A significant reduction in collagen fibers (type I) was found in DT compared to D (p < 0.05). The associated administration of quercetin, glutamine and α-tocopherol increased the levels of circulating interleukin-10 (IL-10) and GSH, and reduced the number of type I collagen fibers. Combined use of systemic quercetin, glutamine and alpha-tocopherol attenuates myocardial fibrosis in diabetic rats.
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Diabetes Mellitus Experimental , Quercetina , Animales , Antioxidantes/metabolismo , Colágeno/metabolismo , Diabetes Mellitus Experimental/metabolismo , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Fibrosis , Glutamina/metabolismo , Glutatión/metabolismo , Interleucina-10/metabolismo , Masculino , Estrés Oxidativo , Quercetina/farmacología , Quercetina/uso terapéutico , Ratas , Ratas Wistar , Superóxido Dismutasa/metabolismo , Factor de Crecimiento Transformador beta/metabolismo , alfa-Tocoferol/farmacología , alfa-Tocoferol/uso terapéuticoRESUMEN
Physiological conditions of low leptin levels like those observed during negative energy balance are usually characterized by the suppression of luteinizing hormone (LH) secretion and fertility. Leptin administration restores LH levels and reproductive function. Leptin action on LH secretion is thought to be mediated by the brain. However, the neuronal population that mediates this effect is still undefined. The hypothalamic ventral premammillary nucleus (PMV) neurons express a dense concentration of leptin receptors and project to brain areas related to reproductive control. Therefore, we hypothesized that the PMV is well located to mediate leptin action on LH secretion. To test our hypothesis, we performed bilateral excitotoxic lesions of the PMV in adult female rats. PMV-lesioned animals displayed a clear disruption of the estrous cycle, remaining in anestrus for 15-20 d. After apparent recovery of cyclicity, animals perfused in the afternoon of proestrus showed decreased Fos immunoreactivity in the anteroventral periventricular nucleus and in gonadotropin releasing hormone neurons. PMV-lesioned animals also displayed decreased estrogen and LH secretion on proestrus. Lesions caused no changes in mean food intake and body weight up to 7 weeks after surgery. We further tested the ability of leptin to induce LH secretion in PMV-lesioned fasted animals. We found that complete lesions of the PMV precluded leptin stimulation of LH secretion on fasting. Our findings demonstrate that the PMV is a key site linking changing levels of leptin and coordinated control of reproduction.
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Ayuno/metabolismo , Leptina/metabolismo , Hormona Luteinizante/metabolismo , Núcleo Hipotalámico Ventromedial/metabolismo , Animales , Ayuno/sangre , Femenino , Leptina/sangre , Hormona Luteinizante/antagonistas & inhibidores , Hormona Luteinizante/sangre , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Reproducción/fisiología , Núcleo Hipotalámico Ventromedial/patologíaRESUMEN
1. Impairmant of baroreflex sensitivity (BRS) has been implicated in the reduction of heart rate variability (HRV) and in the increased risk of death after myocardial infarction (MI). In the present study, we investigated whether the additional impairment in BRS induced by sinoaortic baroreceptor denervation (SAD) in MI rats is associated with changes in the low-frequency (LF) component of HRV and increased mortality rate. 2. Rats were randomly divided into four groups: control, MI, denervated (SAD) and SAD + MI rats. Left ventricular (LV) function was evaluated by echocardiography. Autonomic components were assessed by power spectral analysis and BRS. 3. Myocardial infarction (90 days) reduced ejection fraction (by approximately 42%) in both the MI and SAD + MI groups; however, an increase in LV mass and diastolic dysfunction were observed only in the SAD + MI group. Furthermore, BRS, HRV and the LF power of HRV were reduced after MI, with an exacerbated reduction seen in SAD + MI rats. The LF component of blood pressure variability (BPV) was increased in the MI, SAD and SAD + MI groups compared with the control group. Mortality was higher in the MI groups compared with the non-infarcted groups, with an additional increase in mortality in the SAD + MI group compared with the MI group. Correlations were obtained between BRS and the LF component of HRV and between LV mass and the LF component of BPV. 4. Together, the results indicate that the abolishment of BRS induced by SAD in MI rats further reduces the LF band of HRV, resulting in a worse cardiac remodelling and increased mortality in these rats. These data highlight the importance of this mechanism in the prognosis of patients after an ischaemic event.
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Enfermedades del Sistema Nervioso Autónomo/mortalidad , Enfermedades del Sistema Nervioso Autónomo/fisiopatología , Hemodinámica/fisiología , Infarto del Miocardio/mortalidad , Infarto del Miocardio/fisiopatología , Remodelación Ventricular/fisiología , Animales , Aorta/inervación , Desnervación Autonómica/efectos adversos , Desnervación Autonómica/mortalidad , Enfermedades del Sistema Nervioso Autónomo/complicaciones , Enfermedades del Sistema Nervioso Autónomo/diagnóstico , Barorreflejo/fisiología , Presión Sanguínea/fisiología , Seno Carotídeo/inervación , Frecuencia Cardíaca/fisiología , Hipertrofia Ventricular Izquierda/fisiopatología , Masculino , Infarto del Miocardio/complicaciones , Infarto del Miocardio/diagnóstico , Tamaño de los Órganos , Presorreceptores/cirugía , Pronóstico , Distribución Aleatoria , Ratas , Ratas Wistar , Factores de Riesgo , Volumen Sistólico/fisiología , Nervio Vago/cirugíaRESUMEN
Increased generation of reactive oxygen species (ROS) and altered Ca2+ handling cause vascular damage in hypertension. Mechanisms linking these systems are unclear, but TRPM2 (transient receptor potential melastatin 2) could be important because TRPM2 is a ROS sensor and a regulator of Ca2+ and Na+ transport. We hypothesized that TRPM2 is a point of cross-talk between redox and Ca2+ signaling in vascular smooth muscle cells (VSMC) and that in hypertension ROS mediated-TRPM2 activation increases [Ca2+]i through processes involving NCX (Na+/Ca2+ exchanger). VSMCs from hypertensive and normotensive individuals and isolated arteries from wild type and hypertensive mice (LinA3) were studied. Generation of superoxide anion and hydrogen peroxide (H2O2) was increased in hypertensive VSMCs, effects associated with activation of redox-sensitive PARP1 (poly [ADP-ribose] polymerase 1), a TRPM2 regulator. Ang II (angiotensin II) increased Ca2+ and Na+ influx with exaggerated responses in hypertension. These effects were attenuated by catalase-polyethylene glycol -catalase and TRPM2 inhibitors (2-APB, 8-Br-cADPR olaparib). TRPM2 siRNA decreased Ca2+ in hypertensive VSMCs. NCX inhibitors (Benzamil, KB-R7943, YM244769) normalized Ca2+ hyper-responsiveness and MLC20 phosphorylation in hypertensive VSMCs. In arteries from LinA3 mice, exaggerated agonist (U46619, Ang II, phenylephrine)-induced vasoconstriction was decreased by TRPM2 and NCX inhibitors. In conclusion, activation of ROS-dependent PARP1-regulated TRPM2 contributes to vascular Ca2+ and Na+ influx in part through NCX. We identify a novel pathway linking ROS to Ca2+ signaling through TRPM2/NCX in human VSMCs and suggest that oxidative stress-induced upregulation of this pathway may be a new player in hypertension-associated vascular dysfunction.
Asunto(s)
Señalización del Calcio/fisiología , Hipertensión/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Canales Catiónicos TRPM/metabolismo , Animales , Calcio/metabolismo , Humanos , Peróxido de Hidrógeno/metabolismo , Ratones , Músculo Liso Vascular/metabolismo , Miocitos del Músculo Liso/metabolismo , Oxidación-Reducción , Estrés Oxidativo , Fosforilación , Poli(ADP-Ribosa) Polimerasa-1/metabolismo , Canales Catiónicos TRPM/genéticaRESUMEN
Pulse wave velocity (PWV) has become a gold standard index to quantify the stiffness of the aorta and is a predictor of cardiovascular events. A recent paper compared the pOpmètreâ, a device for measuring the finger-toe PWV, with other techniques and demonstrated its accuracy and validity. However, human devices do not allow the advancement of our knowledge on conditioning mechanisms. Based on its human validation, a new device, pOpetâ 1.0 system was designed for estimation of PWV in small animals and this present study aimed to standardize the pOpetâ 1.0 for estimation of arterial stiffness in rats, and to confirm its liability and stability as well as the reproducibility of assessments. Therefore several precautions were taken into consideration like as the correct position of the animal and photodiodes according to manufacturers' suggestions. Results indicated that estimation of PWV through the new pOpetâ 1.0 device exhibits good internal consistency, stability and objectivity in all tests performed between days and evaluators. Importantly, data suggest for the first time that this new device is able to detect changes in arterial stiffness that are conditioned by age and pressure-related arterial remodeling. ⢠This new pOpetâ device is able to detect changes in vessel structure. ⢠This new pOpetâ device exhibits good internal consistency, stability and objectivity in all tests performed ⢠Correct position of the animal and photodiodes are crucial to obtain a very stable signal.
RESUMEN
AIMS: Transient Receptor Potential Melastatin 7 (TRPM7) cation channel is a chanzyme (channel + kinase) that influences cellular Mg2+ homeostasis and vascular signalling. However, the pathophysiological significance of TRPM7 in the cardiovascular system is unclear. The aim of this study was to investigate the role of this chanzyme in the cardiovascular system focusing on inflammation and fibrosis. METHODS AND RESULTS: TRPM7-deficient mice with deletion of the kinase domain (TRPM7+/Δkinase) were studied and molecular mechanisms investigated in TRPM7+/Δkinase bone marrow-derived macrophages (BMDM) and co-culture systems with cardiac fibroblasts. TRPM7-deficient mice had significant cardiac hypertrophy, fibrosis, and inflammation. Cardiac collagen and fibronectin content, expression of pro-inflammatory mediators (SMAD3, TGFß) and cytokines [interleukin (IL)-6, IL-10, IL-12, tumour necrosis factor-α] and phosphorylation of the pro-inflammatory signalling molecule Stat1, were increased in TRPM7+/Δkinase mice. These processes were associated with infiltration of inflammatory cells (F4/80+CD206+ cardiac macrophages) and increased galectin-3 expression. Cardiac [Mg2+]i, but not [Ca2+]i, was reduced in TRPM7+/Δkinase mice. Calpain, a downstream TRPM7 target, was upregulated (increased expression and activation) in TRPM7+/Δkinase hearts. Vascular functional and inflammatory responses, assessed in vivo by intra-vital microscopy, demonstrated impaired neutrophil rolling, increased neutrophil: endothelial attachment and transmigration of leucocytes in TRPM7+/Δkinase mice. TRPM7+/Δkinase BMDMs had increased levels of galectin-3, IL-10, and IL-6. In co-culture systems, TRPM7+/Δkinase macrophages increased expression of fibronectin, proliferating cell nuclear antigen, and TGFß in cardiac fibroblasts from wild-type mice, effects ameliorated by MgCl2 treatment. CONCLUSIONS: We identify a novel anti-inflammatory and anti-fibrotic role for TRPM7 and suggest that its protective effects are mediated, in part, through Mg2+-sensitive processes.
Asunto(s)
Cardiomegalia/metabolismo , Cardiomiopatías/metabolismo , Mediadores de Inflamación/metabolismo , Inflamación/metabolismo , Miocardio/metabolismo , Canales Catiónicos TRPM/metabolismo , Remodelación Ventricular , Animales , Cardiomegalia/genética , Cardiomegalia/patología , Cardiomegalia/fisiopatología , Cardiomiopatías/genética , Cardiomiopatías/patología , Cardiomiopatías/fisiopatología , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Fibroblastos/metabolismo , Fibroblastos/patología , Fibrosis , Inflamación/genética , Inflamación/patología , Inflamación/fisiopatología , Rodamiento de Leucocito , Macrófagos/metabolismo , Macrófagos/patología , Magnesio/metabolismo , Masculino , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Noqueados , Miocardio/patología , Transducción de Señal , Canales Catiónicos TRPM/deficiencia , Canales Catiónicos TRPM/genética , Migración Transendotelial y TransepitelialRESUMEN
We tested the hypothesis that small changes in angiotensin I-converting enzyme (ACE) expression can alter the vascular response to injury. Male mice containing one, two, three, and four copies of the Ace gene with no detectable vascular abnormality or changes in blood pressure were submitted to cuff-induced femoral artery injury. Femoral thickening was higher in 3- and 4-copy mice (42.4 +/- 4.3% and 45.7 +/- 6.5%, respectively) compared with 1- and 2-copy mice (8.3 +/- 1.3% and 8.5 +/- 0.9%, respectively). Femoral ACE levels from control and injured vessels were assessed in 1- and 3-copy Ace mice, which represent the extremes of the observed response. ACE vascular activity was higher in 3- vs. 1-copy Ace mice (2.4-fold, P < 0.05) in the control uninjured vessel. Upon injury, ACE activity significantly increased in both groups [2.41-fold and 2.14-fold (P < 0.05) for 1- and 3-copy groups, respectively] but reached higher levels in 3- vs. 1-copy Ace mice (P < 0.05). Pharmacological interventions were then used as a counterproof and to indirectly assess the role of angiotensin II (ANG II) on this response. Interestingly, ACE inhibition (enalapril) and ANG II AT(1) receptor blocker (losartan) reduced intima thickening in 3-copy mice to 1-copy mouse values (P < 0.05) while ANG II treatment significantly increased intima thickening in 1-copy mice to 3-copy mouse levels (P < 0.05). Together, these data indicate that small physiologically relevant changes in ACE, not associated with basal vascular abnormalities or blood pressure levels, do influence the magnitude of cuff-induced neointima thickening in mice.
Asunto(s)
Dosificación de Gen , Peptidil-Dipeptidasa A/genética , Túnica Íntima/patología , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Arterias Carótidas/efectos de los fármacos , Arterias Carótidas/enzimología , Arterias Carótidas/patología , Traumatismos de las Arterias Carótidas/fisiopatología , Constricción , Enalapril/farmacología , Arteria Femoral/efectos de los fármacos , Arteria Femoral/enzimología , Arteria Femoral/lesiones , Losartán/farmacología , Ratones , Ratones Transgénicos , Peptidil-Dipeptidasa A/metabolismo , Sistema Renina-Angiotensina/efectos de los fármacos , Sistema Renina-Angiotensina/fisiología , Espectrometría de Fluorescencia , Túnica Íntima/efectos de los fármacos , Túnica Íntima/fisiopatologíaRESUMEN
Angiotensin II plays important functions in cardiovascular system mediating actions leading to inflammatory responses such as activation of VSMC in order to produce ROS, inflammatory cytokines, chemokines, and adhesion molecules. Changes in angiotensin II production could stimulate the recruitment and activation of myeloid cells initiating local inflammatory response without effect on BP. We aimed to verify if angiotensin II induces an inflammatory response in the aorta and if it correlates with variations in BP. C57Bl/6 mice treated with saline solution (0.9%, control group) or angiotensin II (30ng/kg, Ang II group) were used. BP and HR levels were measured. Immunohistochemistry for IL1-ß, TGF-ß, iNOS, CD45, and α-actin was performed in the aorta. BP and HR do not change. A biphasic response was observed both for IL1-ß and TGF-ß expression and also for the presence of CD45 positive cells, with an acute increase (between 30 and 60 minutes) and a second increase, between 24 and 48 hours. Positive staining for iNOS increased in the earlier period (30 minutes) in perivascular adipose tissue and in a longer period (48 hours) in tunica adventitia. Immunoblotting to α-actin showed no alterations, suggesting that the applied dose of angiotensin II does not alter the aortic VSMCs phenotype. The results suggest that angiotensin II, even at doses that do not alter BP, induces the expression of inflammatory markers and migration of inflammatory cells into the aorta of normotensive mice. Thus, angiotensin II may increase the propensity to develop a cardiovascular injury, even in normotensive individuals.
Asunto(s)
Angiotensina II/administración & dosificación , Aorta/efectos de los fármacos , Presión Sanguínea/genética , Enfermedades Cardiovasculares/genética , Actinas/genética , Angiotensina II/efectos adversos , Animales , Aorta/metabolismo , Presión Sanguínea/efectos de los fármacos , Enfermedades Cardiovasculares/inducido químicamente , Enfermedades Cardiovasculares/patología , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Interleucina-1beta/genética , Antígenos Comunes de Leucocito/genética , Ratones , Músculo Liso Vascular/efectos de los fármacos , Músculo Liso Vascular/metabolismo , Óxido Nítrico Sintasa de Tipo II/genética , Solución Salina/administración & dosificación , Factor de Crecimiento Transformador beta/genéticaRESUMEN
This work aimed to evaluate cardiac morphology/function and histological changes induced by bone marrow cells (BMCs) and cultured mesenchymal stem cells (MSCs) injected at the myocardium of spontaneously hypertensive rats (SHR) submitted to surgical coronary occlusion. Female syngeneic adult SHR, submitted (MI) or not (C) to coronary occlusion, were treated 24 h later with in situ injections of normal medium (NM), or with MSCs (MSC) or BMCs (BM) from male rats. The animals were evaluated after 1 and 30 days by echocardiography, histology of heart sections and PCR for the Y chromosome. Improved ejection fraction and reduced left ventricle infarcted area were observed in MSC rats as compared to the other experimental groups. Treated groups had significantly reduced lesion tissue score, increased capillary density and normal (not-atrophied) myocytes, as compared to NM and C groups. The survival rate was higher in C, NM and MSC groups as compared to MI and BM groups. In situ injection of both MSCs and BMCs resulted in improved cardiac morphology, in a more physiological model of myocardial infarction represented by surgical coronary occlusion of spontaneously hypertensive rats. Only treatment with MSCs, however, ameliorated left ventricle dysfunction, suggesting a positive role of these cells in heart remodeling in infarcted hypertensive subjects.
Asunto(s)
Células de la Médula Ósea/citología , Trasplante de Células , Hipertensión/cirugía , Células Madre Mesenquimatosas/citología , Infarto del Miocardio/cirugía , Animales , Secuencia de Bases , Sistema Cardiovascular/fisiopatología , Cartilla de ADN , Modelos Animales de Enfermedad , Ecocardiografía , Femenino , Inmunofenotipificación , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/fisiopatología , Reacción en Cadena de la Polimerasa , Ratas , Ratas Endogámicas SHRRESUMEN
1. Heart regeneration after myocardial infarction (MI) can occur after cell therapy, but the mechanisms, cell types and delivery methods responsible for this improvement are still under investigation. In the present study, we evaluated the impact of systemic delivery of bone marrow cells (BMC) and cultivated mesenchymal stem cells (MSC) on cardiac morphology, function and mortality in spontaneously hypertensive rats (SHR) submitted to coronary occlusion. 2. Female syngeneic adult SHR, submitted or not (control group; C) to MI, were treated with intravenous injection of MSC (MI + MSC) or BMC (MI + BM) from male rats and evaluated after 1, 15 and 30 days by echocardiography. Systolic blood pressure (SBP), functional capacity, histology, mortality rate and polymerase chain reaction for the Y chromosome were also analysed. 3. Myocardial infarction induced a decrease in SBP and BMC, but not MSC, prevented this decrease. An improvement in functional capacity and ejection fraction (38 +/- 4, 39 +/- 3 and 58 +/- 2% for MI, MI + MSC and MI + BM, respectively; P < 0.05), as well as a reduction of the left ventricle infarcted area, were observed in rats from the MI + BM group compared with the other three groups. Treated animals had a significantly reduced lesion tissue score. The mortality rate in the C, MI + BM, MI + MSC and MI groups was 0, 0, 16.7 and 44.4%, respectively (P < 0.05 for the MI + MSC and MI groups compared with the C and MI + BM groups). 4. The results of the present study suggest that systemic administration of BMC can improve left ventricular function, functional capacity and, consequently, reduce mortality in an animal model of MI associated with hypertension. We speculate that the cells transiently home to the myocardium, releasing paracrine factors that recruit host cells to repair the lesion.