RESUMEN
OBJECTIVE: Endothelial outgrowth cells (EOC) decrease inflammation and improve endothelial repair. Inflammation aggravates kidney injury in renal artery stenosis (RAS), and may account for its persistence upon revascularization. We hypothesized that EOC would decrease inflammatory (M1) macrophages and improve renal recovery in RAS. APPROACH AND RESULTS: Pigs with 10 weeks of RAS were studied 4 weeks after percutaneous transluminal renal angioplasty (PTRA+stenting) or sham, with or without adjunct intrarenal delivery of autologous EOC (10×10(6)), and compared with similarly treated normal controls (n=7 each). Single-kidney function, microvascular and tissue remodeling, inflammation, oxidative stress, and fibrosis were evaluated. Four weeks after PTRA, EOC were engrafted in injected RAS-kidneys. Stenotic-kidney glomerular filtration rate was restored in RAS+EOC, RAS+PTRA, and RAS+PTRA+EOC pigs, whereas stenotic-kidney blood flow and angiogenesis were improved and fibrosis attenuated only in EOC-treated pigs. Furthermore, EOC increased cell proliferation and decreased the ratio of M1 (inflammatory)/M2 (reparative) macrophages, as well as circulating levels and stenotic-kidney release of inflammatory cytokines. Cultured-EOC released microvesicles in vitro and induced phenotypic switch (M1-to-M2) in cultured monocytes, which was inhibited by vascular endothelial growth factor blockade. Finally, a single intrarenal injection of rh-vascular endothelial growth factor (0.05 µg/kg) in 7 additional RAS pigs also restored M1/M2 ratio 4 weeks later. CONCLUSIONS: Intrarenal infusion of EOC after PTRA induced a vascular endothelial growth factor-mediated attenuation in macrophages inflammatory phenotype, preserved microvascular architecture and function, and decreased inflammation and fibrosis in the stenotic kidney, suggesting a novel mechanism and therapeutic potential for adjunctive EOC delivery in experimental RAS to improve PTRA outcomes.
Asunto(s)
Células Endoteliales/fisiología , Riñón/fisiopatología , Macrófagos/fisiología , Obstrucción de la Arteria Renal/fisiopatología , Animales , Proliferación Celular , Fibrosis , Hemodinámica , Inflamación/etiología , Riñón/patología , Activación de Macrófagos , Fenotipo , Porcinos , Factor A de Crecimiento Endotelial Vascular/farmacologíaRESUMEN
OBJECTIVES: Magnetic resonance elastography (MRE) can noninvasively sample tissue stiffness in vivo. Renal fibrosis secondary to renal artery stenosis (RAS), which is aggravated in atherosclerotic RAS (ARAS), may increase its stiffness. An increase in cortical stiffness in vivo can be masked by intrinsic hemodynamic determinants, whereas renal medullary stiffness is less dependent on renal hemodynamics. Therefore, this study tested the hypothesis that MRE-determined medullary stiffness would correspond to the histological degree of medullary fibrosis in stenotic kidneys in RAS and detect its exacerbation in ARAS. MATERIALS AND METHODS: Seventeen pigs were studied 10 weeks after induction of unilateral RAS (n = 6), ARAS (n = 5), or sham (n = 6). Stiffness of the cortex and the medulla was determined through 3-dimensional MRE, and renal perfusion and function were determined using multidetector computed tomography. Kidney fibrosis was subsequently assessed ex vivo using the Masson trichrome staining. RESULTS: Renal stenotic cortex and medulla were significantly more fibrotic in RAS and ARAS compared with healthy kidney. However, MRE detected increased stiffness in RAS compared with the healthy kidney (12.7 ± 0.41 kPa vs 10.7 ± 0.18 kPa; P = 0.004) only in the medulla, which was further increased in ARAS (16.6 ± 1.3 kPa; P = 0.017 vs RAS). Magnetic resonance elastography-derived medullary, but not cortical, stiffness significantly correlated with histological degree of fibrosis, although cortical and medullary fibroses were correlated. Renal blood flow and function were similarly decreased in RAS and ARAS compared with the healthy kidney. CONCLUSIONS: Noninvasive 3-dimensional MRE detects increased renal medullary stiffness in RAS and ARAS in vivo, which correlates with its fibrosis ex vivo and may also reflect cortical fibrosis. Hence, MRE-derived medullary stiffness can be potentially useful in detecting renal fibrosis and track disease progression.
Asunto(s)
Diagnóstico por Imagen de Elasticidad , Riñón/irrigación sanguínea , Riñón/patología , Obstrucción de la Arteria Renal/complicaciones , Obstrucción de la Arteria Renal/diagnóstico por imagen , Animales , Femenino , Fibrosis/etiología , PorcinosRESUMEN
AIMS: Humanin (HN) is an endogenous mitochondrial-derived cytoprotective peptide that has shown protective effects against atherosclerosis and is expressed in human vessels. However, its effects on the progression of kidney disease are unknown. We hypothesized that HN would protect the kidney in the early phase of atherogenesis. MAIN METHODS: Forty-eight mice were studied in four groups (n=12 each). Twenty-four ApoE deficient mice were fed a 16-week high-cholesterol diet supplemented with saline or HN (4mg/kg/day, intraperitoneal). C57BL/6 mice were fed a normal diet supplemented with saline or HN. Microvascular architecture was assessed with micro-CT and vascular wall remodeling by alpha-SMA staining. The effects of HN on angiogenesis, inflammation, apoptosis and fibrosis were evaluated in the kidney tissue by Western blotting and histology. KEY FINDINGS: Cortical microvascular spatial density and media/lumen area ratio were significantly increased in high-cholesterol diet fed ApoE deficient mice, but restored by HN. HN up-regulated the renal expressions of anti-angiogenic proteins angiostatin and TSP-1, and inhibited angiopoietin-1. HN attenuated inflammation by down-regulating MCP-1, TNF-alpha and osteopontin. HN also tended to restore pSTAT3 and attenuated Bax expression, suggesting blunted apoptosis. Kidney collagen IV expression was alleviated by HN treatment. SIGNIFICANCE: HN attenuates renal microvascular remodeling, inflammation and apoptosis in the early stage of kidney disease in hypercholesterolemic ApoE(-/-) mice. HN may serve as a novel therapeutic target to mitigate kidney damage in early atherosclerosis.
Asunto(s)
Apolipoproteínas E/genética , Hipercolesterolemia/tratamiento farmacológico , Péptidos y Proteínas de Señalización Intracelular/fisiología , Riñón/efectos de los fármacos , Animales , Apoptosis/efectos de los fármacos , Aterosclerosis/tratamiento farmacológico , Aterosclerosis/fisiopatología , Western Blotting , Progresión de la Enfermedad , Femenino , Hipercolesterolemia/fisiopatología , Inflamación/tratamiento farmacológico , Inflamación/fisiopatología , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Riñón/irrigación sanguínea , Riñón/metabolismo , Enfermedades Renales/tratamiento farmacológico , Enfermedades Renales/etiología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Microvasos/efectos de los fármacos , Neovascularización Patológica/tratamiento farmacológicoRESUMEN
Metabolic syndrome (MetS) is associated with glomerular hyperfiltration and is a risk factor for chronic kidney disease, but the underlying mechanisms are poorly defined. This study tested the hypothesis that increased glomerular filtration rate (GFR) in early MetS is associated with renal adiposity and microvascular proliferation. Twelve MetS-prone Ossabaw pigs were randomized to 10 wk of a standard (lean, n = 6) or atherogenic (MetS, n = 6) diet. Kidney hemodynamics and function, perirenal fat volume, and tubular dynamics were assessed in vivo by multidetector computed tomography (CT) and blood oxygen level-dependent (BOLD)-MRI. Microvascular architecture was assessed ex vivo with micro-CT. Candidate injury mechanisms were evaluated in kidney tissue by Western blotting and histology. Basal GFR, renal blood flow, and renal cortical perfusion and volume were elevated in the MetS group. Perirenal and kidney tissue fat, proximal-nephron intratubular fluid concentration, and endothelial nitric oxide synthase expression were increased in MetS. GFR levels correlated with tissue triglyceride levels. Elevated spatial density of 20- to 40-µm cortical microvessels was accompanied by mild oxidative stress, inflammation, and with proximal tubular vacuolization. Medullary size and perfusion were relatively preserved, and BOLD-MRI showed intact medullary tubular response to furosemide. Increased GFR in early MetS is associated with renal adiposity and microvascular proliferation, which involve mainly the renal cortex and precede significant activation of oxidative stress and inflammation. Renal adiposity and proliferative microvessels may represent novel therapeutic targets for preserving renal function in early MetS.
Asunto(s)
Adiposidad/fisiología , Capilares/fisiología , Tasa de Filtración Glomerular/fisiología , Riñón/fisiopatología , Síndrome Metabólico/fisiopatología , Animales , Presión Sanguínea/fisiología , Proliferación Celular , Dieta , Genotipo , Inflamación/fisiopatología , Riñón/metabolismo , Pruebas de Función Renal , Túbulos Renales/fisiología , Imagen por Resonancia Magnética , Síndrome Metabólico/patología , Neovascularización Fisiológica/fisiología , Estrés Oxidativo/fisiología , Oxígeno/sangre , Circulación Renal/efectos de los fármacos , Porcinos , Tomografía Computarizada por Rayos X , Triglicéridos/metabolismoRESUMEN
OBJECTIVES: : Magnetic resonance elastography (MRE) allows noninvasive assessment of tissue stiffness in vivo. Renal arterial stenosis (RAS), a narrowing of the renal artery, promotes irreversible tissue fibrosis that threatens kidney viability and may elevate tissue stiffness. However, kidney stiffness may also be affected by hemodynamic factors. This study tested the hypothesis that renal blood flow (RBF) is an important determinant of renal stiffness as measured by MRE. MATERIAL AND METHODS: : In 6 anesthetized pigs MRE studies were performed to determine cortical and medullary elasticity during acute graded decreases in RBF (by 20%, 40%, 60%, 80%, and 100% of baseline) achieved by a vascular occluder. Three sham-operated swine served as time control. Additional pigs were studied with MRE 6 weeks after induction of chronic unilateral RAS (n = 6) or control (n = 3). Kidney fibrosis was subsequently evaluated histologically by trichrome staining. RESULTS: : During acute RAS the stenotic cortex stiffness decreased (from 7.4 ± 0.3 to 4.8 ± 0.6 kPa, P = 0.02 vs. baseline) as RBF decreased. Furthermore, in pigs with chronic RAS (80% ± 5.4% stenosis) in which RBF was decreased by 60% ± 14% compared with controls, cortical stiffness was not significantly different from normal (7.4 ± 0.3 vs. 7.6 ± 0.3 kPa, P = 0.3), despite histologic evidence of renal tissue fibrosis. CONCLUSION: : Hemodynamic variables modulate kidney stiffness measured by MRE and may mask the presence of fibrosis. These results suggest that kidney turgor should be considered during interpretation of elasticity assessments.