RESUMO
BACKGROUND: Acute kidney injury affects ~70% of asphyxiated newborns, and increases their risk of developing chronic kidney disease later in life. Acute kidney injury is driven by renal oxygen deprivation during asphyxia, thus we hypothesized that creatine administered antenatally would protect the kidney from the long-term effects of birth asphyxia. METHODS: Pregnant spiny mice were fed standard chow or chow supplemented with 5% creatine from 20-d gestation (midgestation). One day prior to term (37-d gestation), pups were delivered by caesarean or subjected to intrauterine asphyxia. Litters were allocated to one of two time-points. Kidneys were collected at 1 mo of age to estimate nephron number (stereology). Renal function (excretory profile and glomerular filtration rate) was measured at 3 mo of age, and kidneys then collected for assessment of glomerulosclerosis. RESULTS: Compared with controls, at 1 mo of age male (but not female) birth-asphyxia offspring had 20% fewer nephrons (P < 0.05). At 3 mo of age male birth-asphyxia offspring had 31% lower glomerular filtration rate (P < 0.05) and greater glomerular collagen IV content (P < 0.01). Antenatal creatine prevented these renal injuries arising from birth asphyxia. CONCLUSION: Maternal creatine supplementation during pregnancy may be an effective prophylactic to prevent birth asphyxia induced acute kidney injury and the emergence of chronic kidney disease.
Assuntos
Injúria Renal Aguda/prevenção & controle , Asfixia Neonatal/fisiopatologia , Creatina/uso terapêutico , Rim/fisiopatologia , Fenômenos Fisiológicos da Nutrição Materna , Injúria Renal Aguda/fisiopatologia , Animais , Animais Recém-Nascidos , Colágeno Tipo IV/metabolismo , Creatina/administração & dosagem , Suplementos Nutricionais , Feminino , Falência Renal Crônica/fisiopatologia , Falência Renal Crônica/prevenção & controle , Glomérulos Renais/fisiopatologia , Masculino , Camundongos , Néfrons/fisiopatologia , Tamanho do Órgão , Oxigênio/metabolismo , Gravidez , PrenhezRESUMO
We assessed the utility of synchrotron-radiation micro-computed tomography (micro-CT) for quantification of the radial geometry of the renal cortical vasculature. The kidneys of nine rats and six rabbits were perfusion fixed and the renal circulation filled with Microfil. In order to assess shrinkage of Microfil, rat kidneys were imaged at the Australian Synchrotron immediately upon tissue preparation and then post fixed in paraformaldehyde and reimaged 24 hours later. The Microfil shrank only 2-5% over the 24 hour period. All subsequent micro-CT imaging was completed within 24 hours of sample preparation. After micro-CT imaging, the kidneys were processed for histological analysis. In both rat and rabbit kidneys, vascular structures identified in histological sections could be identified in two-dimensional (2D) micro-CT images from the original kidney. Vascular morphology was similar in the two sets of images. Radial geometry quantified by manual analysis of 2D images from micro-CT was consistent with corresponding data generated by light microscopy. However, due to limited spatial resolution when imaging a whole organ using contrast-enhanced micro-CT, only arteries ≥100 and ≥60 µm in diameter, for the rat and rabbit respectively, could be assessed. We conclude that it is feasible and valid to use micro-CT to quantify vascular geometry of the renal cortical circulation in both the rat and rabbit. However, a combination of light microscopic and micro-CT approaches are required to evaluate the spatial relationships between intrarenal arteries and veins over an extensive range of vessel size.
Assuntos
Angiografia por Tomografia Computadorizada/métodos , Rim/diagnóstico por imagem , Microscopia/métodos , Artéria Renal/diagnóstico por imagem , Veias Renais/diagnóstico por imagem , Animais , Interpretação de Imagem Assistida por Computador , Técnicas In Vitro , Rim/irrigação sanguínea , Coelhos , Ratos , Especificidade da EspécieRESUMO
Oxygen tension (Po2) of urine in the bladder could be used to monitor risk of acute kidney injury if it varies with medullary Po2 Therefore, we examined this relationship and characterized oxygen diffusion across walls of the ureter and bladder in anesthetized rabbits. A computational model was then developed to predict medullary Po2 from bladder urine Po2 Both intravenous infusion of [Phe(2),Ile(3),Orn(8)]-vasopressin and infusion of N(G)-nitro-l-arginine reduced urinary Po2 and medullary Po2 (8-17%), yet had opposite effects on renal blood flow and urine flow. Changes in bladder urine Po2 during these stimuli correlated strongly with changes in medullary Po2 (within-rabbit r(2) = 0.87-0.90). Differences in the Po2 of saline infused into the ureter close to the kidney could be detected in the bladder, although this was diminished at lesser ureteric flow. Diffusion of oxygen across the wall of the bladder was very slow, so it was not considered in the computational model. The model predicts Po2 in the pelvic ureter (presumed to reflect medullary Po2) from known values of bladder urine Po2, urine flow, and arterial Po2 Simulations suggest that, across a physiological range of urine flow in anesthetized rabbits (0.1-0.5 ml/min for a single kidney), a change in bladder urine Po2 explains 10-50% of the change in pelvic urine/medullary Po2 Thus, it is possible to infer changes in medullary Po2 from changes in urinary Po2, so urinary Po2 may have utility as a real-time biomarker of risk of acute kidney injury.
Assuntos
Testes de Função Renal/métodos , Medula Renal/metabolismo , Modelos Biológicos , Oxigênio/urina , Bexiga Urinária/metabolismo , Micção/fisiologia , Animais , Simulação por Computador , Oxigênio/sangue , Coelhos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Recent evidence obtained from a rodent model of birth asphyxia shows that supplementation of the maternal diet with creatine during pregnancy protects the neonate from multi-organ damage. However, the effect of increasing creatine intake on creatine homeostasis and biosynthesis in females, particularly during pregnancy, is unknown. This study assessed the impact of creatine supplementation on creatine homeostasis, body composition, capacity for de novo creatine synthesis and renal excretory function in non-pregnant and pregnant spiny mice. Mid-gestation pregnant and virgin spiny mice were fed normal chow or chow supplemented with 5 % w/w creatine for 18 days. Weight gain, urinary creatine and electrolyte excretion were assessed during supplementation. At post mortem, body composition was assessed by Dual-energy X-ray absorptiometry, or tissues were collected to assess creatine content and mRNA expression of the creatine synthesising enzymes arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT) and the creatine transporter (CrT1). Protein expression of AGAT and GAMT was also assessed by Western blot. Key findings of this study include no changes in body weight or composition with creatine supplementation; increased urinary creatine excretion in supplemented spiny mice, with increased sodium (P < 0.001) and chloride (P < 0.05) excretion in pregnant dams after 3 days of supplementation; lowered renal AGAT mRNA (P < 0.001) and protein (P < 0.001) expressions, and lowered CrT1 mRNA expression in the kidney (P < 0.01) and brain (P < 0.001). Creatine supplementation had minimal impact on creatine homeostasis in either non-pregnant or pregnant spiny mice. Increasing maternal dietary creatine consumption could be a useful treatment for birth asphyxia.
Assuntos
Creatina , Suplementos Nutricionais , Homeostase/efeitos dos fármacos , Rim/metabolismo , Amidinotransferases/biossíntese , Animais , Creatina/farmacocinética , Creatina/farmacologia , Feminino , Guanidinoacetato N-Metiltransferase/biossíntese , Homeostase/fisiologia , Testes de Função Renal , Proteínas de Membrana Transportadoras/metabolismo , Camundongos , GravidezRESUMO
PURPOSE: Epidemiological and experimental studies demonstrate that intrauterine growth restriction (IUGR) followed by accelerated postnatal growth leads to increased risk of developing cardiac disease in adulthood. The aim of this study was to examine the effect of early life growth restriction on cardiac structure and function in young adult rats. METHODS: IUGR was induced in Wistar Kyoto dams through administration of a low protein diet (LPD; 8.7% casein) during pregnancy and lactation; controls received a normal protein diet (NPD; 20% casein). Cardiac function and structure were assessed in female NPD (n = 7) and LPD (n = 7) offspring at 18 weeks of age by echocardiography and pressure-volume techniques, and systolic blood pressure by tail-cuff sphygmomanometry. RESULTS: LPD offspring remained significantly smaller throughout life compared to controls. There were no differences in the levels of systolic blood pressure, left ventricular cardiac dimensions, heart rate, ejection fraction and fractional shortening of the cardiac muscle between the investigated groups. Aortic peak systolic velocity was significantly reduced in the LPD group (P = 0.02). CONCLUSION: Our findings support the idea that the programming of adult cardiovascular disease can be prevented or delayed in IUGR offspring when postnatal growth trajectory resembles that of in utero.
Assuntos
Retardo do Crescimento Fetal/fisiopatologia , Ventrículos do Coração/fisiopatologia , Fenômenos Fisiológicos da Nutrição Materna , Animais , Pressão Sanguínea , Doenças Cardiovasculares/prevenção & controle , Dieta com Restrição de Proteínas/efeitos adversos , Feminino , Retardo do Crescimento Fetal/etiologia , Frequência Cardíaca , Ventrículos do Coração/embriologia , Lactação , Masculino , Miocárdio/metabolismo , Gravidez , Ratos , Ratos Endogâmicos WKYRESUMO
BACKGROUND: Pregnancy induces adaptations in maternal metabolism to meet the increased need for nutrients by the placenta and fetus. Creatine is an important intracellular metabolite obtained from the diet and also synthesised endogenously. Experimental evidence suggests that the fetus relies on a maternal supply of creatine for much of gestation. However, the impact of pregnancy on maternal creatine homeostasis is unclear. We hypothesise that alteration of maternal creatine homeostasis occurs during pregnancy to ensure adequate levels of this essential substrate are available for maternal tissues, the placenta and fetus. This study aimed to describe maternal creatine homeostasis from mid to late gestation in the precocial spiny mouse. METHODS: Plasma creatine concentration and urinary excretion were measured from mid to late gestation in pregnant (n = 8) and age-matched virgin female spiny mice (n = 6). At term, body composition and organ weights were assessed and tissue total creatine content determined. mRNA expression of the creatine synthesising enzymes arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), and the creatine transporter (CrT1) were assessed by RT-qPCR. Protein expression of AGAT and GAMT was also assessed by western blot analysis. RESULTS: Plasma creatine and renal creatine excretion decreased significantly from mid to late gestation (P < 0.001, P < 0.05, respectively). Pregnancy resulted in increased lean tissue (P < 0.01), kidney (P < 0.01), liver (P < 0.01) and heart (P < 0.05) mass at term. CrT1 expression was increased in the heart (P < 0.05) and skeletal muscle (P < 0.05) at term compared to non-pregnant tissues, and creatine content of the heart (P < 0.05) and kidney (P < 0.001) were also increased at this time. CrT1 mRNA expression was down-regulated in the liver (<0.01) and brain (<0.01) of pregnant spiny mice at term. Renal AGAT mRNA (P < 0.01) and protein (P < 0.05) expression were both significantly up-regulated at term, with decreased expression of AGAT mRNA (<0.01) and GAMT protein (<0.05) observed in the term pregnant heart. Brain AGAT (<0.01) and GAMT (<0.001) mRNA expression were also decreased at term. CONCLUSION: Change of maternal creatine status (increased creatine synthesis and reduced creatine excretion) may be a necessary adjustment of maternal physiology to pregnancy to meet the metabolic demands of maternal tissues, the placenta and developing fetus.
Assuntos
Amidinotransferases/genética , Creatina/metabolismo , Guanidinoacetato N-Metiltransferase/genética , Homeostase/genética , Proteínas de Membrana Transportadoras/genética , Gravidez/metabolismo , RNA Mensageiro/metabolismo , Amidinotransferases/metabolismo , Animais , Western Blotting , Feminino , Regulação da Expressão Gênica , Guanidinoacetato N-Metiltransferase/metabolismo , Murinae , Gravidez/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Rodent models of renal physiology and pathology are crucial to our understanding of the molecular, histological and functional sequelae that contribute to kidney diseases. One of the most important measures of renal function is glomerular filtration rate (GFR). While the accurate determination of GFR is pivotal to understanding the progression of disease and/or the benefits of treatment strategies, in rodents the conventional methods for assessment of GFR are inconvenient and cumbersome, not the least because they involve stress and often anaesthesia. The legitimacy of assay-based assessment of plasma and urine markers of GFR in mice has also been heavily scrutinized for their insensitivity to minor declines in GFR and inaccurate detection of renal biomarkers. While infusion-based clearance methods of GFR assessment are thus the gold standard in terms of accuracy, they are limited by the fact that they are primarily non-recovery procedures. This presents a dilemma when trying to document the progression of renal disease, as these measures cannot be taken in the same experimental subject. Here we review a technique of transcutaneous measurement of fluorescein isothiocyanate-labelled sinistrin to calculate GFR in small rodents, using a non-invasive clearance device (NIC-Kidney Device). This is a recently validated non-invasive technique for measuring GFR in small rodents that allows for the real-time measurement of GFR in conscious animals, without the need for plasma and urine assays.
Assuntos
Tamanho Corporal , Taxa de Filtração Glomerular , Rim/fisiologia , Animais , Fluoresceínas/farmacocinética , Corantes Fluorescentes/farmacocinética , Rim/metabolismo , Camundongos Endogâmicos C57BL , Modelos Animais , Modelos Biológicos , Oligossacarídeos/farmacocinética , Reprodutibilidade dos Testes , Espectrometria de FluorescênciaRESUMO
Renal arterial-to-venous (AV) oxygen shunting limits oxygen delivery to renal tissue. To better understand how oxygen in arterial blood can bypass renal tissue, we quantified the radial geometry of AV pairs and how it differs according to arterial diameter and anatomic location. We then estimated diffusion of oxygen in the vicinity of arteries of typical geometry using a computational model. The kidneys of six rats were perfusion fixed, and the vasculature was filled with silicone rubber (Microfil). A single section was chosen from each kidney, and all arteries (n = 1,628) were identified. Intrarenal arteries were largely divisible into two "types," characterized by the presence or absence of a close physical relationship with a paired vein. Arteries with a close physical relationship with a paired vein were more likely to have a larger rather than smaller diameter, and more likely to be in the inner-cortex than the mid- or outer cortex. Computational simulations indicated that direct diffusion of oxygen from an artery to a paired vein can only occur when the two vessels have a close physical relationship. However, even in the absence of this close relationship oxygen can diffuse from an artery to periarteriolar capillaries and venules. Thus AV oxygen shunting in the proximal preglomerular circulation is dominated by direct diffusion of oxygen to a paired vein. In the distal preglomerular circulation, it may be sustained by diffusion of oxygen from arteries to capillaries and venules close to the artery wall, which is subsequently transported to renal veins by convection.
Assuntos
Difusão , Córtex Renal/irrigação sanguínea , Modelos Biológicos , Oxigênio/sangue , Animais , Masculino , Ratos Sprague-Dawley , Circulação RenalRESUMO
Sex hormones regulate the renin-angiotensin system. For example, estrogen enhances expression of the angiotensin type 2 receptor. We hypothesized that activation of the angiotensin type 2 receptor shifts the chronic pressure-natriuresis relationship leftward in females compared with males and that this effect is lost with age. Mean arterial pressure was measured by radiotelemetry in adult (4 mo old) and aged (14 mo old) wild-type and angiotensin type 2 receptor knockout male and female mice. Chronic pressure-natriuresis curves were constructed while mice were maintained on a normal-salt (0.26%) diet and following 6 days of high salt (5.0%) diet. Mean arterial pressure was lower in adult wild-type females than males (88 ± 1 and 97 ± 1 mmHg, respectively), a difference that was maintained with age, but was absent in adult knockout mice. In wild-type females, the chronic pressure-natriuresis relationship was shifted leftward compared with knockout females, an effect that was lost with age. In males, the chronic pressure-natriuresis relationship was not influenced by angiotensin type 2 receptor deficiency. Compared with age-matched females, the chronic pressure-natriuresis relationships in male mice were shifted rightward. Renal expression of the angiotensin type 2 receptor was fourfold greater in adult wild-type females than males. With age, the angiotensin type 2 receptor-to-angiotensin type 1 receptor balance was reduced in females. Conversely, in males, angiotensin receptor expression did not vary significantly with age. In conclusion, the angiotensin type 2 receptor modulates the chronic pressure-natriuresis relationship in an age- and sex-dependent manner.
Assuntos
Natriurese/fisiologia , Receptor Tipo 2 de Angiotensina/fisiologia , Fatores Etários , Animais , Pressão Sanguínea , Feminino , Masculino , Camundongos , Camundongos Knockout , Receptor Tipo 2 de Angiotensina/genética , Receptor Tipo 2 de Angiotensina/metabolismo , Fatores SexuaisRESUMO
BACKGROUND: Acute kidney injury (AKI) is a major complication for infants following an asphyxic insult at birth. We aimed to determine if kidney structure and function were affected in an animal model of birth asphyxia and if maternal dietary creatine supplementation could provide an energy reserve to the fetal kidney, maintaining cellular respiration during asphyxia and preventing AKI. METHODS: Pregnant spiny mice were maintained on normal chow or chow supplemented with creatine from day 20 gestation. On day 38 (term ~39 d), pups were delivered by cesarean section (c-section) or subjected to intrauterine asphyxia. Twenty-four hours after insult, kidneys were collected for histological or molecular analysis. Urine and plasma were also collected for biochemical analysis. RESULTS: AKI was evident at 24 h after birth asphyxia, with a higher incidence of shrunken glomeruli (P < 0.02), disturbance to tubular arrangement, tubular dilatation, a twofold increase (P < 0.02) in expression of Ngal (early marker of kidney injury), and decreased expression of the podocyte differentiation marker nephrin. Maternal creatine supplementation prevented the glomerular and tubular abnormalities observed in the kidney at 24 h and the increased expression of Ngal. CONCLUSION: Maternal creatine supplementation may prove useful in ameliorating kidney injury associated with birth asphyxia.
Assuntos
Injúria Renal Aguda/prevenção & controle , Asfixia Neonatal/tratamento farmacológico , Creatina/administração & dosagem , Suplementos Nutricionais , Rim/efeitos dos fármacos , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/genética , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Injúria Renal Aguda/fisiopatologia , Animais , Asfixia Neonatal/complicações , Biomarcadores/metabolismo , Citoproteção , Modelos Animais de Doenças , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Idade Gestacional , Humanos , Recém-Nascido , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Lipocalinas/metabolismo , Proteínas de Membrana/metabolismo , Murinae , Gravidez , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
While low nephron number is associated with increased risk of developing cardiovascular and renal disease, the functional consequences of a high nephron number are unknown. We tested the hypothesis that a high nephron number provides protection against hypertensive and renal insults. Mean arterial pressure (MAP) and renal function were characterized in male wild-type (WT) and transforming growth factor-ß2 heterozygous (Tgfb2(+/-)) mice under basal conditions and following a chronic high-salt diet. Kidneys were collected for unbiased stereological analysis. Baseline MAP and renal function were indistinguishable between genotypes. The chronic high-salt diet (5% NaCl for 4 wk followed by 8% NaCl for 4 wk) led to similar step-wise increases in urine volume, Na(+) excretion, and albuminuria in the genotypes. The 5% NaCl diet induced modest and similar increases in MAP (3.5 ± 1.6 and 3.4 ± 0.8 mmHg in WT and Tgfb2(+/-), respectively). After the step up to the 8% NaCl diet, MAP increased further in WT (+15.9 ± 5.1 mmHg), but not Tgfb2(+/-) (-0.1 ± 1.0 mmHg), mice. Nephron number was 30% greater in Tgfb2(+/-) than WT mice and was not affected by the chronic high-salt diet. Mean glomerular volume was lower in Tgfb2(+/-) than WT mice, and the chronic high-salt diet induced significant glomerular hypertrophy. In a separate cohort of mice, an acute, 7-day, 8% NaCl diet induced similar rises in MAP in the genotypes. This is the first study to examine the physiological characteristics of a model of high nephron number, and the findings are consistent with this phenotype providing protection against chronic, but not acute, hypertensive insults.
Assuntos
Hipertensão/induzido quimicamente , Hipertensão/prevenção & controle , Néfrons/citologia , Néfrons/fisiologia , Cloreto de Sódio/efeitos adversos , Animais , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Contagem de Células , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Genótipo , Heterozigoto , Hipertensão/patologia , Masculino , Camundongos , Camundongos Mutantes , Néfrons/efeitos dos fármacos , Fenótipo , Cloreto de Sódio/farmacologia , Fator de Crescimento Transformador beta2/genética , Fator de Crescimento Transformador beta2/fisiologiaRESUMO
Colony-stimulating factor (CSF)-1 controls the survival, proliferation, and differentiation of macrophages, which are recognized as scavengers and agents of the innate and the acquired immune systems. Because of their plasticity, macrophages are endowed with many other essential roles during development and tissue homeostasis. We present evidence that CSF-1 plays an important trophic role in postnatal organ growth and kidney repair. Notably, the injection of CSF-1 postnatally enhanced kidney weight and volume and was associated with increased numbers of tissue macrophages. Moreover, CSF-1 promotes postnatal renal repair in mice after ischemia-reperfusion injury by recruiting and influencing macrophages toward a reparative state. CSF-1 treatment rapidly accelerated renal repair with tubular epithelial cell replacement, attenuation of interstitial fibrosis, and functional recovery. Analysis of macrophages from CSF-1-treated kidneys showed increased expression of insulin-like growth factor-1 and anti-inflammatory genes that are known CSF-1 targets. Taken together, these data suggest that CSF-1 is important in kidney growth and the promotion of endogenous repair and resolution of inflammatory injury.
Assuntos
Rim/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Macrófagos/efeitos dos fármacos , Traumatismo por Reperfusão/prevenção & controle , Injúria Renal Aguda/patologia , Injúria Renal Aguda/fisiopatologia , Injúria Renal Aguda/prevenção & controle , Animais , Animais Recém-Nascidos , Peso Corporal/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Colágeno/metabolismo , Perfilação da Expressão Gênica , Rim/irrigação sanguínea , Rim/metabolismo , Rim/patologia , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Tamanho do Órgão/efeitos dos fármacos , Fenótipo , Recuperação de Função Fisiológica , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/fisiopatologiaRESUMO
Development of the kidney can be altered in utero in response to a suboptimal environment. The intrarenal factors that have been most well characterized as being sensitive to programming events are kidney mass/nephron endowment, the renin-angiotensin system, tubular sodium handling, and the renal sympathetic nerves. Newborns that have been subjected to an adverse intrauterine environment may thus begin life at a distinct disadvantage, in terms of renal function, at a time when the kidney must take over the primary role for extracellular fluid homeostasis from the placenta. A poor beginning, causing renal programming, has been linked to increased risk of hypertension and renal disease in adulthood. However, although a cause for concern, increasingly, evidence demonstrates that renal programming is not a fait accompli in terms of future cardiovascular and renal disease. A greater understanding of postnatal renal maturation and the impact of secondary factors (genes, sex, diet, stress, and disease) on this process is required to predict which babies are at risk of increased cardiovascular and renal disease as adults and to be able to devise preventative measures.
Assuntos
Rim/embriologia , Rim/fisiopatologia , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Animais , Doenças Cardiovasculares/epidemiologia , Feminino , Homeostase/fisiologia , Humanos , Nefropatias/epidemiologia , Gravidez , Fatores de RiscoRESUMO
The extent to which a reduced nephron endowment contributes to hypertension and renal disease is confounded in models created by intrauterine insults that also demonstrate other phenotypes. Furthermore, recent data suggest that a reduced nephron endowment provides the "first hit" and simply increases the susceptibility to injurious stimuli. Thus we examined nephron number, glomerular volume, conscious mean arterial pressure (MAP), and renal function in a genetic model of reduced nephron endowment before and after a high-salt (5%) diet. One-yr-old glial cell line-derived neurotrophic factor wild-type (WT) mice, heterozygous (HET) mice born with two kidneys (HET2K), and HET mice born with one kidney (HET1K) were used. Nephron number was 25% lower in HET2K and 65% lower in HET1K than WT mice. Glomeruli hypertrophied in both HET groups by 33%, resulting in total glomerular volumes that were similar between HET2K and WT mice but remained 50% lower in HET1K mice. On a normal-salt diet, 24-h MAP was not different between WT, HET2K, and HET1K mice (102 +/- 1, 103 +/- 1, and 102 +/- 2 mmHg). On a high-salt diet, MAP increased 9.1 +/- 1.9 mmHg in HET1K mice (P < 0.05) and 5.4 +/- 0.9 mmHg in HET2K mice (P < 0.05) and did not change significantly in WT mice. Creatinine clearance was 60% higher in WT mice but 30% lower in HET2K and HET1K mice fed a high-salt diet than in controls maintained on a normal-salt diet. Thus a reduction in nephron number (or total glomerular volume) alone does not lead to hypertension or kidney disease in aged mice, but exposure to high salt uncovers a hypertensive and renal phenotype.
Assuntos
Pressão Sanguínea , Hipertensão/etiologia , Nefropatias/etiologia , Néfrons/fisiopatologia , Cloreto de Sódio na Dieta/efeitos adversos , Animais , Pressão Sanguínea/genética , Ritmo Circadiano , Creatinina/sangue , Modelos Animais de Doenças , Feminino , Genótipo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/genética , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Taxa de Filtração Glomerular , Heterozigoto , Hipertensão/genética , Hipertensão/metabolismo , Hipertensão/patologia , Hipertensão/fisiopatologia , Hipertrofia , Nefropatias/genética , Nefropatias/metabolismo , Nefropatias/patologia , Nefropatias/fisiopatologia , Testes de Função Renal , Glomérulos Renais/anormalidades , Glomérulos Renais/metabolismo , Glomérulos Renais/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Néfrons/anormalidades , Néfrons/metabolismo , Concentração Osmolar , Fenótipo , Renina/sangue , Sódio/sangue , MicçãoRESUMO
AIMS: Renal inflammation, leading to fibrosis and impaired function is a major contributor to the development of hypertension. The NLRP3 inflammasome mediates inflammation in several chronic diseases by processing the cytokines pro-interleukin (IL)-1ß and pro-IL-18. In this study, we investigated whether MCC950, a recently-identified inhibitor of NLRP3 activity, reduces blood pressure (BP), renal inflammation, fibrosis and dysfunction in mice with established hypertension. METHODS AND RESULTS: C57BL6/J mice were made hypertensive by uninephrectomy and treatment with deoxycorticosterone acetate (2.4 mg/day, s.c.) and 0.9% NaCl in the drinking water (1K/DOCA/salt). Normotensive controls were uninephrectomized and received normal drinking water. Ten days later, mice were treated with MCC950 (10 mg/kg/day, s.c.) or vehicle (saline, s.c.) for up to 25 days. BP was monitored by tail-cuff or radiotelemetry; renal function by biochemical analysis of 24-h urine collections; and kidney inflammation/pathology was assessed by real-time PCR for inflammatory gene expression, flow cytometry for leucocyte influx, and Picrosirius red histology for collagen. Over the 10 days post-surgery, 1K/DOCA/salt-treated mice became hypertensive, developed impaired renal function, and displayed elevated renal levels of inflammatory markers, collagen and immune cells. MCC950 treatment from day 10 attenuated 1K/DOCA/salt-induced increases in renal expression of inflammasome subunits (NLRP3, ASC, pro-caspase-1) and inflammatory/injury markers (pro-IL-18, pro-IL-1ß, IL-17A, TNF-α, osteopontin, ICAM-1, VCAM-1, CCL2, vimentin), each by 25-40%. MCC950 reduced interstitial collagen and accumulation of certain leucocyte subsets in kidneys of 1K/DOCA/salt-treated mice, including CD206+ (M2-like) macrophages and interferon-gamma-producing T cells. Finally, MCC950 partially reversed 1K/DOCA/salt-induced elevations in BP, urine output, osmolality, [Na+], and albuminuria (each by 20-25%). None of the above parameters were altered by MCC950 in normotensive mice. CONCLUSION: MCC950 was effective at reducing BP and limiting renal inflammation, fibrosis and dysfunction in mice with established hypertension. This study provides proof-of-concept that pharmacological inhibition of the NLRP3 inflammasome is a viable anti-hypertensive strategy.
Assuntos
Anti-Inflamatórios/farmacologia , Anti-Hipertensivos/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Furanos/farmacologia , Hipertensão/prevenção & controle , Rim/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Cloreto de Sódio na Dieta , Sulfonamidas/farmacologia , Albuminúria/etiologia , Albuminúria/metabolismo , Albuminúria/fisiopatologia , Albuminúria/prevenção & controle , Animais , Quimiotaxia de Leucócito/efeitos dos fármacos , Colágeno/metabolismo , Acetato de Desoxicorticosterona , Modelos Animais de Doenças , Fibrose , Compostos Heterocíclicos de 4 ou mais Anéis , Hipertensão/etiologia , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Indenos , Mediadores da Inflamação/metabolismo , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nefrectomia , Transdução de Sinais , Sulfonas , Subpopulações de Linfócitos T/efeitos dos fármacos , Subpopulações de Linfócitos T/metabolismoRESUMO
The cellular origins of glomerulosclerosis involve activation of parietal epithelial cells (PECs) and progressive podocyte depletion. While mammalian target of rapamycin-mediated (mTOR-mediated) podocyte hypertrophy is recognized as an important signaling pathway in the context of glomerular disease, the role of podocyte hypertrophy as a compensatory mechanism preventing PEC activation and glomerulosclerosis remains poorly understood. In this study, we show that glomerular mTOR and PEC activation-related genes were both upregulated and intercorrelated in biopsies from patients with focal segmental glomerulosclerosis (FSGS) and diabetic nephropathy, suggesting both compensatory and pathological roles. Advanced morphometric analyses in murine and human tissues identified podocyte hypertrophy as a compensatory mechanism aiming to regulate glomerular functional integrity in response to somatic growth, podocyte depletion, and even glomerulosclerosis - all of this in the absence of detectable podocyte regeneration. In mice, pharmacological inhibition of mTOR signaling during acute podocyte loss impaired hypertrophy of remaining podocytes, resulting in unexpected albuminuria, PEC activation, and glomerulosclerosis. Exacerbated and persistent podocyte hypertrophy enabled a vicious cycle of podocyte loss and PEC activation, suggesting a limit to its beneficial effects. In summary, our data highlight a critical protective role of mTOR-mediated podocyte hypertrophy following podocyte loss in order to preserve glomerular integrity, preventing PEC activation and glomerulosclerosis.
Assuntos
Albuminúria/induzido quimicamente , Nefropatias Diabéticas/patologia , Everolimo/efeitos adversos , Glomerulosclerose Segmentar e Focal/patologia , Serina-Treonina Quinases TOR/metabolismo , Idoso , Idoso de 80 Anos ou mais , Animais , Biópsia , Células Cultivadas , Pré-Escolar , Conjuntos de Dados como Assunto , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/patologia , Nefropatias Diabéticas/tratamento farmacológico , Células Epiteliais/patologia , Everolimo/administração & dosagem , Feminino , Perfilação da Expressão Gênica , Humanos , Hipertrofia/tratamento farmacológico , Hipertrofia/patologia , Lactente , Masculino , Camundongos , Camundongos Knockout , Pessoa de Meia-Idade , Podócitos , Cultura Primária de Células , Regeneração , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Estreptozocina/toxicidade , Serina-Treonina Quinases TOR/análise , Serina-Treonina Quinases TOR/antagonistas & inibidores , Proteína 1 do Complexo Esclerose Tuberosa/genética , Proteína 1 do Complexo Esclerose Tuberosa/metabolismo , Regulação para Cima , Adulto JovemRESUMO
The influence of endogenous endothelins on the neural control of renal function is poorly understood. We therefore studied the effects of endothelin blockade (combined ET(A) and ET(B) receptor antagonism using TAK-044) on the acute and prolonged effects of renal nerve stimulation in rabbits, measuring renal blood flow, glomerular filtration rate (GFR), urine flow and sodium excretion. Brief (3 min) stimulation over 0.5-8 Hz produced frequency-dependent reductions in total renal blood flow, cortical blood flow and, less markedly, medullary blood flow. TAK-044 did not significantly alter basal total renal blood flow or cortical blood flow, or their responses to nerve stimulation, but significantly increased basal medullary blood flow (P<0.01) and increased the slope of the stimulation frequency-medullary blood flow relationship (P<0.05). Prolonged (20 min) stimulation at 0, 0.5 and 2 Hz produced frequency-dependent reductions in total renal blood flow, GFR, urine flow and sodium excretion, but not medullary blood flow. Pretreatment with TAK-044 did not significantly alter these responses. Thus, endogenous endothelins do not appear to either augment or lessen the effects of renal nerve activation on total renal blood flow, cortical blood flow, GFR or sodium excretion. The apparent ability of TAK-044 to enhance medullary blood flow responses to renal nerve stimulation may reflect an action of endogenous endothelins to blunt neurally mediated vasoconstriction in the medullary circulation. Alternatively, it may simply be secondary to the effects of endogenous endothelins on basal medullary blood flow.
Assuntos
Endotelinas/metabolismo , Rim/irrigação sanguínea , Rim/inervação , Rim/metabolismo , Circulação Renal/fisiologia , Anestesia , Animais , Estimulação Elétrica , Antagonistas dos Receptores de Endotelina , Taxa de Filtração Glomerular/efeitos dos fármacos , Masculino , Peptídeos Cíclicos/farmacologia , Coelhos , Circulação Renal/efeitos dos fármacosRESUMO
Recently, a combined probe has been developed capable of simultaneous measurement of local tissue pO2 (fluorescence oximetry) and microvascular perfusion (laser Doppler flux) within the same local region. The aim of the current study was to test the utility of these combined probes to measure pO2 and perfusion in the kidney. Studies were performed in anesthetized, artificially ventilated rabbits (n=7). Baseline measurements of renal medullary perfusion and pO2 obtained using combined probes (537 +/- 110 units & 28.7 +/- 6.l mmHg, respectively) were indistinguishable from those obtained using independent probes (435 +/- 102 units & 26.9 +/- 6.4 mmHg). Baseline measurements of renal cortical pO2 were also similar between combined (9.7 +/- 1.6 mmHg) and independent probes (9.5 +/- 2.3 mmHg). Baseline levels of cortical perfusion however, were significantly greater when measured using independent probes (1130 +/- 114 units) compared to combined probes (622 +/- 59 units; P < 0.02). Relative changes in perfusion and pO2 resulting from graded stimulation of the renal nerves were not significantly different when measured using combined probes to those obtained using independent probes. We conclude that combined probes are equally suitable to independent probes for tissue pO2 and microvascular perfusion measurement in the kidney. Our results raise some concerns regarding the accuracy of these OxyLite fluorescence probes for pO2 measurement in the kidney, particularly within the renal cortex.
Assuntos
Rim/metabolismo , Fluxometria por Laser-Doppler , Oximetria , Oxigênio/metabolismo , Animais , Hemodinâmica , Rim/irrigação sanguínea , Fluxometria por Laser-Doppler/instrumentação , Fluxometria por Laser-Doppler/métodos , Masculino , Oximetria/instrumentação , Oximetria/métodos , Perfusão , Coelhos , Fluxo Sanguíneo RegionalRESUMO
OBJECTIVE: To investigate the role of endothelin in noradrenaline-induced hypertension in rats. DESIGN: The dose-response relationship of chronic noradrenaline infusion on arterial pressure was characterized to identify a dose that would produce sustained hypertension, and the effect of combined endothelin ETA and ETB receptor blockade (TAK-044) on the response to this dose was then examined. METHODS AND RESULTS: Noradrenaline (or vehicle) was infused intravenously at 1 (subpressor acutely), 24 or 48 microg/kg per h (acute pressor response of 9 +/- 1 and 11 +/- 1 mmHg, respectively) for a 14-day infusion, and blood pressure was measured by radiotelemetry. Noradrenaline infusion at 1 microg/kg per h did not produce a 'slow pressor' rise in blood pressure. During noradrenaline infusions at 24 and 48 microg/kg per h, mean arterial pressure peaked initially on days 2-3 (+10 +/- 1 and 14 +/- 2 mmHg, respectively; P < 0.01), fell towards basal levels after day 3, and then began to rise again at days 5-6 only with 48 microg/kg per h, being 10 +/- 1 mmHg above control levels at days 13-14 (P < 0.05). TAK-044 treatment did not alter the magnitude of the initial (13 +/- 1 mmHg) or eventual (12 +/- 2 mmHg) rise in blood pressure achieved in response to 14 days' infusion of noradrenaline at 48 microg/kg per h, but abolished the transient fall. CONCLUSION: Chronic noradrenaline infusion at acutely pressor doses leads either to a transient blood pressure elevation at a moderate dose, or to a triphasic but sustained hypertension at a higher dose, with a temporary escape from the hypertension apparently mediated by endothelin.
Assuntos
Endotelinas/fisiologia , Hipertensão/induzido quimicamente , Norepinefrina/farmacologia , Animais , Pressão Sanguínea/efeitos dos fármacos , Relação Dose-Resposta a Droga , Frequência Cardíaca/efeitos dos fármacos , Masculino , Peptídeos Cíclicos/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor de Endotelina B/fisiologiaRESUMO
OBJECTIVE: To determine the effects of chronic denervation on renal vascular structure and function in young adult spontaneously hypertensive rats (SHRs). DESIGN: Unilateral renal denervation (SHRUDx) or sham-operation (SHRS) was performed in SHRs at 6 weeks of age. At 10 weeks, rats were allocated to one of three procedures designed to examine renal vascular structure and function. A further group underwent bilateral renal denervation. METHODS: In SHRUDx or SHRS groups, either the kidneys were perfusion-fixed for stereological estimates of artery wall and lumen dimensions or for vascular casting to determine arteriole lumen diameters, or the rats were anaesthetized for estimation of glomerular capillary pressure. RESULTS: Chronic unilateral renal denervation had no significant effect on the development of hypertension between 6 and 10 weeks of age, as previously reported, but resulted in luminal narrowing of the interlobular artery (denervated group 52 +/- 2 mum, sham-operated group 64 +/- 1 mum; P < 0.01 for interaction between strain and treatment), without alterations in interlobular or arcuate artery wall dimensions. There were no significant effects on either afferent or efferent arteriole lumen diameters. Estimated glomerular capillary pressure was significantly lower in the denervated kidneys of SHRUDx (47 +/- 1 mmHg) compared with kidneys of the SHRS (50 +/- 1 mmHg; P < 0.04). Mean arterial pressure was approximately 12 mmHg lower in the bilaterally denervated SHRs than in the sham-operated SHRs. CONCLUSIONS: Although bilateral denervation attenuated the development of hypertension in SHRs, unilateral denervation did not, indicating that one neurally intact kidney was sufficient to drive the normal development of SHR hypertension, but only with apparent prohypertensive compensatory changes in the denervated kidney.