RESUMEN
PURPOSE: Euterpe oleracea Mart. (açaí) seed extract (ASE), through its anti-hypertensive, antioxidant and anti-inflammatory properties, may be useful to treat or prevent human diseases. Several evidences suggest that oxidative stress and inflammation contribute to the pathogenesis of diabetic nephropathy; therefore, we tested the hypothesis that ASE (200 mg/kg-1day-1) prevents diabetes and hypertension-related oxidative stress and inflammation, attenuating renal injury. METHODS: Male rats with streptozotocin (STZ)-induced diabetes (D), and spontaneously hypertensive rats with STZ-induced diabetes (DH) were treated daily with tap water or ASE (D + ASE and DH + ASE, respectively) for 45 days. The control (C) and hypertensive (H) animals received water. RESULTS: The elevated serum levels of urea and creatinine in D and DH, and increased albumin excretion in HD were reduced by ASE. Total glomeruli number in D and DH, were increased by ASE that also reduced renal fibrosis in both groups by decreasing collagen IV and TGF-ß1 expression. ASE improved biomarkers of renal filtration barrier (podocin and nephrin) in D and DH groups and prevented the increased expression of caspase-3, IL-6, TNF-α and MCP-1 in both groups. ASE reduced oxidative damage markers (TBARS, carbonyl levels and 8-isoprostane) in D and DH associated with a decrease in Nox 4 and p47 subunit expression and increase in antioxidant enzyme activity in both groups (SOD, catalase and GPx). CONCLUSION: ASE substantially reduced renal injury and prevented renal dysfunction by reducing inflammation, oxidative stress and improving the renal filtration barrier, providing a nutritional resource for prevention of diabetic and hypertensive-related nephropathy.
Asunto(s)
Antioxidantes/uso terapéutico , Nefropatías Diabéticas/prevención & control , Suplementos Dietéticos , Euterpe/química , Extractos Vegetales/uso terapéutico , Insuficiencia Renal/prevención & control , Semillas/química , Animales , Antiinflamatorios no Esteroideos/uso terapéutico , Antihipertensivos/uso terapéutico , Apoptosis , Biomarcadores/sangre , Biomarcadores/metabolismo , Biomarcadores/orina , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/dietoterapia , Diabetes Mellitus Experimental/inmunología , Nefropatías Diabéticas/complicaciones , Nefropatías Diabéticas/metabolismo , Nefropatías Diabéticas/patología , Fibrosis , Barrera de Filtración Glomerular/inmunología , Barrera de Filtración Glomerular/metabolismo , Barrera de Filtración Glomerular/patología , Barrera de Filtración Glomerular/fisiopatología , Hipertensión/complicaciones , Hipertensión/dietoterapia , Hipertensión/inmunología , Hipertensión/fisiopatología , Mediadores de Inflamación/sangre , Mediadores de Inflamación/metabolismo , Riñón/inmunología , Riñón/metabolismo , Riñón/patología , Riñón/fisiopatología , Estrés Oxidativo , Ratas Endogámicas SHR , Insuficiencia Renal/complicaciones , Insuficiencia Renal/etiología , Insuficiencia Renal/metabolismoRESUMEN
The glomerular filtration barrier, has historically only been spatially resolved using electron microscopy due to the nanometer-scale dimensions of these structures. Recently, it was shown that the nanoscale distribution of proteins in the slit diaphragm can be resolved by fluorescence based stimulated emission depletion microscopy, in combination with optical clearing. Fluorescence microscopy has advantages over electron microscopy in terms of multiplex imaging of different epitopes, and also the amount of volumetric data that can be extracted from thicker samples. However, stimulated emission depletion microscopy is still a costly technique commonly not available to most life science researchers. An imaging technique with which the glomerular filtration barrier can be visualized using more standard fluorescence imaging techniques is thus desirable. Recent studies have shown that biological tissue samples can be isotropically expanded, revealing nanoscale localizations of multiple epitopes using confocal microscopy. Here we show that kidney samples can be expanded sufficiently to study the finest elements of the filtration barrier using confocal microscopy. Thus, our result opens up the possibility to study protein distributions and foot process morphology on the effective nanometer-scale.
Asunto(s)
Barrera de Filtración Glomerular/patología , Glomerulonefritis/patología , Microscopía Confocal , Microscopía Fluorescente , Expansión de Tejido/métodos , Animales , Autoanticuerpos , Biomarcadores/metabolismo , Colágeno Tipo IV/inmunología , Colágeno Tipo IV/metabolismo , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente Indirecta , Barrera de Filtración Glomerular/inmunología , Barrera de Filtración Glomerular/metabolismo , Glomerulonefritis/inmunología , Glomerulonefritis/metabolismo , Péptidos y Proteínas de Señalización Intracelular/inmunología , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/inmunología , Proteínas de la Membrana/metabolismo , Ratones Transgénicos , RatasRESUMEN
AIMS: Growing evidences suggest that acute hyperglycemia is strongly related to kidney injury. Our study aimed to investigate the effects of acute hyperglycemia on kidney glomerular and tubular impairment in non-diabetic conscious rats. METHODS: Non-diabetic conscious rats were randomly subjected to 6h of saline (control group) or high glucose (acute hyperglycemia group) infusion. Blood glucose was maintained at 16.0-18.0 mmol/L in acute hyperglycemia group. Renal structure and function alterations, systemic/renal inflammation and oxidative stress markers were assessed, and apoptosis markers of renal inherent cells were evaluated. RESULTS: Acute hyperglycemia caused significant injury to structure of glomerular filtration barrier, tubular epithelial cells and peritubular vascular endothelial cells. It increased urinary microalbumin (68.01 ± 27.09 µg/24h vs 33.81 ± 13.81 µg/24h , P=0.014), ß2-microglobulin, Cystatin C, urinary and serous neutrophil gelatinase-associated lipocalin levels (P < 0.05). Acute hyperglycemia decreased megalin and cubilin expression, activated systemic and renal oxidative stress as well as inflammation and promoted renal inherent cell apoptosis. CONCLUSIONS: Acute hyperglycemia causes significant injury to kidney function and structure. Compared with damages of glomerular filtration barrier, renal tubular injury may contribute more to acute hyperglycemia induced proteinuria. Activation of inflammation especially renal inflammation, oxidative stress and enhanced apoptosis may be the underlying mechanisms.
Asunto(s)
Apoptosis , Hiperglucemia/fisiopatología , Túbulos Renales/fisiopatología , Estrés Oxidativo , Insuficiencia Renal/etiología , Animales , Biomarcadores/sangre , Biomarcadores/metabolismo , Biomarcadores/orina , Glucemia/análisis , Barrera de Filtración Glomerular/inmunología , Barrera de Filtración Glomerular/metabolismo , Barrera de Filtración Glomerular/fisiopatología , Barrera de Filtración Glomerular/ultraestructura , Técnica de Clampeo de la Glucosa , Hiperglucemia/inmunología , Hiperglucemia/metabolismo , Hiperglucemia/patología , Glomérulos Renales/inmunología , Glomérulos Renales/metabolismo , Glomérulos Renales/fisiopatología , Glomérulos Renales/ultraestructura , Túbulos Renales/inmunología , Túbulos Renales/metabolismo , Túbulos Renales/ultraestructura , Masculino , Microscopía Electrónica de Transmisión , Nefritis/etiología , Especificidad de Órganos , Proteinuria/etiología , Distribución Aleatoria , Ratas Sprague-Dawley , Insuficiencia Renal/fisiopatología , Índice de Severidad de la EnfermedadRESUMEN
Podocytes are essential to the structure and function of the glomerular filtration barrier; however, they also exhibit increased expression of MHC class II molecules under inflammatory conditions, and they remove Ig and immune complexes from the glomerular basement membrane (GBM). This finding suggests that podocytes may act as antigen-presenting cells, taking up and processing antigens to initiate specific T cell responses, similar to professional hematopoietic cells such as dendritic cells or macrophages. Here, MHC-antigen complexes expressed exclusively on podocytes of transgenic mice were sufficient to activate CD8+ T cells in vivo. In addition, deleting MHC class II exclusively on podocytes prevented the induction of experimental anti-GBM nephritis. Podocytes ingested soluble and particulate antigens, activated CD4+ T cells, and crosspresented exogenous antigen on MHC class I molecules to CD8+ T cells. In conclusion, podocytes participate in the antigen-specific activation of adaptive immune responses, providing a potential target for immunotherapies of inflammatory kidney diseases and transplant rejection.
Asunto(s)
Células Presentadoras de Antígenos/citología , Células Presentadoras de Antígenos/inmunología , Barrera de Filtración Glomerular/inmunología , Podocitos/citología , Podocitos/inmunología , Inmunidad Adaptativa/inmunología , Animales , Enfermedad por Anticuerpos Antimembrana Basal Glomerular/inmunología , Enfermedad por Anticuerpos Antimembrana Basal Glomerular/patología , Presentación de Antígeno/inmunología , Linfocitos T CD8-positivos/citología , Linfocitos T CD8-positivos/inmunología , Comunicación Celular/inmunología , Femenino , Membrana Basal Glomerular/citología , Membrana Basal Glomerular/inmunología , Membrana Basal Glomerular/metabolismo , Rechazo de Injerto/inmunología , Rechazo de Injerto/patología , Antígenos de Histocompatibilidad Clase II/inmunología , Antígenos de Histocompatibilidad Clase II/metabolismo , Inmunoglobulina G/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos CBA , Ratones Noqueados , Microesferas , Ovalbúmina/inmunología , Ovalbúmina/farmacocinética , Fagocitosis/inmunología , Podocitos/metabolismoRESUMEN
AIMS/HYPOTHESIS: High intraglomerular pressure causes renal inflammation in experimental models of diabetes. Our objective was to determine whether renal hyperfiltration, a surrogate for intraglomerular hypertension, is associated with increased excretion of urinary cytokines/chemokines in patients with type 1 diabetes mellitus. METHODS: Blood pressure, renal haemodynamic function (inulin and para-aminohippurate clearances for glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), respectively) and urine samples were obtained during clamped euglycaemia in individuals with type 1 diabetes with either hyperfiltration (GFR determined using inulin [GFRINULIN] ≥ 135 ml min⻹ 1.73 m⻲, n = 28) or normofiltration (n = 21) and healthy control individuals (n = 18). RESULTS: Baseline clinical characteristics, dietary sodium and protein intake and blood pressure levels were similar in the diabetic and healthy control groups. In addition, HbA1c levels were similar in the two diabetic groups. As expected baseline GFR was higher in hyperfilterers than either normofiltering diabetic patients or healthy control patients (165 ± 9 vs 113 ± 2 and 116 ± 4 ml min⻹ 1.73 m⻲, respectively, p < 0.01). ERPF and renal blood flow were also comparatively higher and renal vascular resistance was lower in hyperfiltering patients (p < 0.01). Hyperfiltering diabetic patients had higher excretion rates for eotaxin, IFNα2, macrophage-derived chemokine, platelet-derived growth factor (PDGF)-AA, PDGF-AB/BB and granulocyte-macrophage colony-stimulating factor (p ≤ 0.01). Urinary monocyte chemoattractant protein (MCP)-1 and RANTES (regulated on activation, normal T expressed and secreted) excretion was also higher in hyperfiltering vs normofiltering diabetic individuals (p < 0.01) and fibroblast growth factor-2, MCP-3 and CD40K excretion was elevated in hyperfiltering diabetic individuals vs healthy controls (p < 0.01). CONCLUSIONS/INTERPRETATION: Renal hyperfiltration is associated with increased urinary excretion of inflammatory cytokines/chemokines in patients with uncomplicated type 1 diabetes.