RESUMEN
Hypertension is a leading risk factor for stroke, heart disease, chronic kidney disease, vascular cognitive impairment, and Alzheimer's disease. Previous genetic studies have nominated hundreds of genes linked to hypertension, and renal and cognitive diseases. Some have been advanced as candidate genes by showing that they can alter blood pressure or renal and cerebral vascular function in knockout animals; however, final validation of the causal variants and underlying mechanisms has remained elusive. This review chronicles 40 years of work, from the initial identification of adducin (ADD) as an ACTIN-binding protein suggested to increase blood pressure in Milan hypertensive rats, to the discovery of a mutation in ADD1 as a candidate gene for hypertension in rats that were subsequently linked to hypertension in man. More recently, a recessive K572Q mutation in ADD3 was identified in Fawn-Hooded Hypertensive (FHH) and Milan Normotensive (MNS) rats that develop renal disease, which is absent in resistant strains. ADD3 dimerizes with ADD1 to form functional ADD protein. The mutation in ADD3 disrupts a critical ACTIN-binding site necessary for its interactions with actin and spectrin to regulate the cytoskeleton. Studies using Add3 KO and transgenic strains, as well as a genetic complementation study in FHH and MNS rats, confirmed that the K572Q mutation in ADD3 plays a causal role in altering the myogenic response and autoregulation of renal and cerebral blood flow, resulting in increased susceptibility to hypertension-induced renal disease and cerebral vascular and cognitive dysfunction.
Asunto(s)
Proteínas de Unión a Calmodulina/genética , Predisposición Genética a la Enfermedad/genética , Hipertensión Renal/genética , Hipertensión/genética , Nefritis/genética , Medicina de Precisión/métodos , Animales , Presión Sanguínea/genética , Disfunción Cognitiva/genética , Modelos Animales de Enfermedad , Homeostasis/genética , Humanos , Mutación , Medicina de Precisión/tendencias , Ratas , Circulación Renal/genéticaRESUMEN
We have reported previously that renal hemodynamic abnormalities exist in the prediabetic stage of type II diabetic rats. At this prediabetic stage these rats have hyperinsulinemia, insulin resistance and metabolic syndrome. It is well known that insulin resistance is frequently associated with renal abnormalities, but the mechanism underlying this association has remained speculative. Although insulin is known to modify renal hemodynamics, little is known about the roles of insulin receptor substrates (IRS1, IRS2) in the renal actions of insulin. To address this issue, the effects of insulin on renal function and renal hemodynamics were investigated in C57BL/6 (WT: wild type), insulin receptor substrate 1- knockout (IRS1-/-), and IRS2-knockout (IRS2-/-) mice. IRS2-/-mice had elevated glucose level as expected. 24-h urine collections and serum creatinine revealed that creatinine clearance did not significantly differ between these groups. Albuminuria was found in IRS1-/-and IRS2-/-groups. We examined the effects on the IRS during the administration of Losartan, which is widely used for diabetic nephropathy. After the administration of Losartan the IRS displayed improved renal hemodynamics. Moreover, the subjects were also given Pioglitazone, which improves insulin resistance. Losartan significantly reduced albuminuria in both groups. Pioglitazone also showed similar results. We assessed the autoregulatory responses of the total renal blood flow (RBF), the superficial (SBF) and the deep renal cortical blood flow (DBF) with stepwise reductions of renal perfusion pressure (RPP), which was induced by a manual clamp on the abdominal aorta. During the clamp induced reductions of the RPP by 10 to 20mm HG, RBF, SBF and the DBF fell significantly more in the IRS1 and IRS2 than in the WT mice. Furthermore micropuncture studies showded that compared to the WT tubuloglomerular feedback (TGF) responses of the stop flow pressure (Psf) were reduced in both the IRS1 -/- and IRS2 -/-. The results of the IRS1 and IRS2 mice displayed the pressence of hemodynamic abnormalities. Losartan and Pioglitazone have shown the potential to improve these abnormalities. In conclusion the results indicate that IRS plays a major role in the stimulation of renal functions and renal hemodynamics in type type II diabetes.
Asunto(s)
Diabetes Mellitus Tipo 2/genética , Nefropatías Diabéticas/genética , Hiperinsulinismo/genética , Proteínas Sustrato del Receptor de Insulina/genética , Albuminuria/sangre , Albuminuria/genética , Animales , Glucemia/genética , Creatinina/sangre , Diabetes Mellitus Tipo 2/sangre , Diabetes Mellitus Tipo 2/patología , Nefropatías Diabéticas/sangre , Nefropatías Diabéticas/patología , Monitorización Hemodinámica/métodos , Hemodinámica , Hiperinsulinismo/sangre , Hiperinsulinismo/patología , Insulina/sangre , Resistencia a la Insulina/genética , Riñón/irrigación sanguínea , Riñón/metabolismo , Riñón/patología , Masculino , Ratones , Ratones Noqueados , Estado Prediabético/sangre , Estado Prediabético/genética , Estado Prediabético/patología , Ratas , Circulación Renal/genética , Transducción de Señal/genéticaRESUMEN
BACKGROUND: The genes and mechanisms involved in the association between diabetes or hypertension and CKD risk are unclear. Previous studies have implicated a role for γ-adducin (ADD3), a cytoskeletal protein encoded by Add3. METHODS: We investigated renal vascular function in vitro and in vivo and the susceptibility to CKD in rats with wild-type or mutated Add3 and in genetically modified rats with overexpression or knockout of ADD3. We also studied glomeruli and primary renal vascular smooth muscle cells isolated from these rats. RESULTS: This study identified a K572Q mutation in ADD3 in fawn-hooded hypertensive (FHH) rats-a mutation previously reported in Milan normotensive (MNS) rats that also develop kidney disease. Using molecular dynamic simulations, we found that this mutation destabilizes a critical ADD3-ACTIN binding site. A reduction of ADD3 expression in membrane fractions prepared from the kidney and renal vascular smooth muscle cells of FHH rats was associated with the disruption of the F-actin cytoskeleton. Compared with renal vascular smooth muscle cells from Add3 transgenic rats, those from FHH rats had elevated membrane expression of BKα and BK channel current. FHH and Add3 knockout rats exhibited impairments in the myogenic response of afferent arterioles and in renal blood flow autoregulation, which were rescued in Add3 transgenic rats. We confirmed these findings in a genetic complementation study that involved crossing FHH and MNS rats that share the ADD3 mutation. Add3 transgenic rats showed attenuation of proteinuria, glomerular injury, and kidney fibrosis with aging and mineralocorticoid-induced hypertension. CONCLUSIONS: This is the first report that a mutation in ADD3 that alters ACTIN binding causes renal vascular dysfunction and promotes the susceptibility to kidney disease.
Asunto(s)
Proteínas de Unión a Calmodulina/genética , Hipertensión/complicaciones , Enfermedades Renales/etiología , Mutación/efectos de los fármacos , Circulación Renal/genética , Animales , Modelos Animales de Enfermedad , Homeostasis , Hipertensión/genética , Glomérulos Renales/metabolismo , Glomérulos Renales/patología , Masculino , Músculo Liso Vascular/fisiología , Miocitos del Músculo Liso/metabolismo , Ratas , Ratas Sprague-Dawley , Ratas TransgénicasRESUMEN
20-Hydroxy-5,8,11,14-eicosatetraenoic acid (20-HETE) has been linked to pro-hypertensive and anti-hypertensive actions through its ability to promote vasoconstriction and inhibit Na transport in the ascending limb of the loop of Henle, respectively. In this study, we assessed the effects of 20-HETE blockade on blood pressure, renal hemodynamics, and urinary sodium excretion in Cyp4a14(-/-) male mice, which display androgen-driven 20-HETE-dependent hypertension. Administration of 2,5,8,11,14,17-hexaoxanonadecan-19-yl 20-hydroxyicosa-6(Z),15(Z)-dienoate (20-SOLA), a water-soluble 20-HETE antagonist, in the drinking water normalized the blood pressure of male Cyp4a14(-/-) hypertensive mice (±124 vs. ±153 mmHg) while having no effect on age-matched normotensive wild-type (WT) male mice. Hypertension in Cyp4a14(-/-) male mice was accompanied by decreased renal perfusion and reduced glomerular filtration rates, which were corrected by treatment with 20-SOLA. Interestingly, Cyp4a14(-/-) male mice treated with 20-SOLA displayed increased urinary sodium excretion that was paralleled by the reduction of blood pressure suggestive of an antinatriuretic activity of endogenous 20-HETE in the hypertensive mice. This interpretation is in line with the observation that the natriuretic response to acute isotonic saline loading in hypertensive Cyp4a14(-/-) male mice was significantly impaired relative to that in WT mice; this impairment was corrected by 20-SOLA treatment. Hence, endogenous 20-HETE appears to promote sodium conservation in hypertensive Cyp4a14(-/-) male mice, presumably, as a result of associated changes in renal hemodynamics and/or direct stimulatory action on tubular sodium reabsorption.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Familia 4 del Citocromo P450/genética , Ácidos Hidroxieicosatetraenoicos/antagonistas & inhibidores , Natriuresis/efectos de los fármacos , Animales , Antihipertensivos/farmacología , Femenino , Tasa de Filtración Glomerular/genética , Túbulos Renales/metabolismo , Masculino , Ratones , Ratones Noqueados , Polietilenglicoles , Circulación Renal/genética , Siloxanos , Sodio/orinaRESUMEN
Countercurrent systems have evolved in a variety of biological systems that allow transfer of heat, gases, and solutes. For example, in the renal medulla, the countercurrent arrangement of vascular and tubular elements facilitates the trapping of urea and other solutes in the inner medulla, which in turn enables the formation of concentrated urine. Arteries and veins in the cortex are also arranged in a countercurrent fashion, as are descending and ascending vasa recta in the medulla. For countercurrent diffusion to occur, barriers to diffusion must be small. This appears to be characteristic of larger vessels in the renal cortex. There must also be gradients in the concentration of molecules between afferent and efferent vessels, with the transport of molecules possible in either direction. Such gradients exist for oxygen in both the cortex and medulla, but there is little evidence that large gradients exist for other molecules such as carbon dioxide, nitric oxide, superoxide, hydrogen sulfide, and ammonia. There is some experimental evidence for arterial-to-venous (AV) oxygen shunting. Mathematical models also provide evidence for oxygen shunting in both the cortex and medulla. However, the quantitative significance of AV oxygen shunting remains a matter of controversy. Thus, whereas the countercurrent arrangement of vasa recta in the medulla appears to have evolved as a consequence of the evolution of Henle's loop, the evolutionary significance of the intimate countercurrent arrangement of blood vessels in the renal cortex remains an enigma.
Asunto(s)
Evolución Biológica , Gases/sangre , Riñón/irrigación sanguínea , Riñón/fisiología , Circulación Renal/genética , Urea/sangre , Animales , Transporte Biológico Activo/genética , Humanos , Arteria Renal , Venas RenalesRESUMEN
In male rats, androgen supplements increase 20-hydroxyeicosatetraenoic acid (20-HETE) via cytochrome P-450 (CYP)4A ω-hydroxylase and cause an increase in blood pressure (BP). In the present study, we determined the roles of 20-HETE and CYP4A2 on the elevated BP in hyperandrogenemic female rats. Chronic dihydrotestosterone (DHT) increased mean arterial pressure (MAP) in female Sprague-Dawley rats (96 ± 2 vs. 108 ± 2 mmHg, P < 0.05) and was associated with increased renal microvascular CYP4A2 mRNA expression (15-fold), endogenous renal 20-HETE (5-fold), and ω-hydroxylase activity (3-fold). Chronic DHT also increased MAP in low salt-fed Dahl salt-resistant female rats (81 ± 4 vs. 95 ± 1 mmHg, P < 0.05) but had no effect on MAP in Dahl salt-sensitive female rats (154 ± 3 vs. 153 ± 3 mmHg), which are known to be 20-HETE deficient. To test the role of CYP4A2, female CYP4A2(-/-) and SS.5(Bn) (wild type) rats were treated with DHT. DHT increased MAP in SS.5(Bn) female rats (104 ± 1 vs. 128 ± 1 mmHg, P < 0.05) but had no effect in CYP4A2(-/-) female rats (118 ± 1 vs. 120 ± 1 mmHg). Renal microvascular 20-HETE was reduced in control CYP4A2(-/-) female rats and was increased with DHT in SS.5(Bn) female rats (6-fold) but not CYP4A2(-/-) female rats. ω-Hydroxylase activity was 40% lower in control CYP4A2(-/-) female rats than in SS.5(Bn) female rats, and DHT decreased ω-hydroxylase activity in SS.5(Bn) female rats (by 50%) but significantly increased ω-hydroxylase activity in CYP4A2(-/-) female rats (3-fold). These data suggest that 20-HETE via CYP4A2 contributes to the elevation in BP in hyperandrogenemic female rats. The data also suggest that 20-HETE synthesis inhibition may be effective in treating the elevated BP in women with hyperandrogenemia, such as women with polycystic ovary syndrome.
Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Ácidos Hidroxieicosatetraenoicos/metabolismo , Hiperandrogenismo/metabolismo , Hipertensión/metabolismo , Andrógenos/toxicidad , Animales , Peso Corporal/genética , Sistema Enzimático del Citocromo P-450/genética , Dihidrotestosterona/toxicidad , Femenino , Eliminación de Gen , Técnicas de Inactivación de Genes , Ácidos Hidroxieicosatetraenoicos/genética , Hiperandrogenismo/genética , Hipertensión/genética , Microcirculación/genética , Ratas , Ratas Endogámicas Dahl , Circulación Renal/genética , Esteroides/sangreRESUMEN
Blood filtration in the kidney glomerulus is essential for physiological homeostasis. The filtration apparatus of the kidney glomerulus is composed of three distinct components: the fenestrated endothelial cells, the glomerular basement membrane, and interdigitating foot processes of podocytes that form the slit diaphragm. Recent studies have demonstrated that podocytes play a crucial role in blood filtration and in the pathogenesis of proteinuria and glomerular sclerosis; however, the molecular mechanisms that organize the podocyte filtration barrier are not fully understood. In this study, we suggest that tight junction protein 1 (Tjp1 or ZO-1), which is encoded by Tjp1 gene, plays an essential role in establishing the podocyte filtration barrier. The podocyte-specific deletion of Tjp1 down-regulated the expression of podocyte membrane proteins, impaired the interdigitation of the foot processes and the formation of the slit diaphragm, resulting in glomerular dysfunction. We found the possibility that podocyte filtration barrier requires the integration of two independent units, the pre-existing epithelial junction components and the newly synthesized podocyte-specific components, at the final stage in glomerular morphogenesis, for which Tjp1 is indispensable. Together with previous findings that Tjp1 expression was decreased in glomerular diseases in human and animal models, our results indicate that the suppression of Tjp1 could directly aggravate glomerular disorders, highlights Tjp1 as a potential therapeutic target.
Asunto(s)
Células Endoteliales/metabolismo , Podocitos/metabolismo , Circulación Renal/genética , Proteína de la Zonula Occludens-1/biosíntesis , Animales , Células Endoteliales/patología , Regulación de la Expresión Génica , Membrana Basal Glomerular/metabolismo , Humanos , Glomérulos Renales/metabolismo , Proteínas de la Membrana/biosíntesis , Ratones , Ratones Noqueados , Podocitos/patología , Proteína de la Zonula Occludens-1/genéticaRESUMEN
Smooth muscle contraction is activated primarily by phosphorylation at Ser19 of the regulatory light chain subunits (LC20) of myosin II, catalysed by Ca(2+)/calmodulin-dependent myosin light chain kinase. Ca(2+)-independent contraction can be induced by inhibition of myosin light chain phosphatase, which correlates with diphosphorylation of LC20 at Ser19 and Thr18, catalysed by integrin-linked kinase (ILK) and zipper-interacting protein kinase (ZIPK). LC20 diphosphorylation at Ser19 and Thr18 has been detected in mammalian vascular smooth muscle tissues in response to specific contractile stimuli (e.g. endothelin-1 stimulation of rat renal afferent arterioles) and in pathophysiological situations associated with hypercontractility (e.g. cerebral vasospasm following subarachnoid hemorrhage). Comparison of the effects of LC 20 monophosphorylation at Ser19 and diphosphorylation at Ser19 and Thr18 on contraction and relaxation of Triton-skinned rat caudal arterial smooth muscle revealed that phosphorylation at Thr18 has no effect on steady-state force induced by Ser19 phosphorylation. On the other hand, the rates of dephosphorylation and relaxation are significantly slower following diphosphorylation at Thr18 and Ser19 compared to monophosphorylation at Ser19. We propose that this diphosphorylation mechanism underlies the prolonged contractile response of particular vascular smooth muscle tissues to specific stimuli, e.g. endothelin-1 stimulation of renal afferent arterioles, and the vasospastic behavior observed in pathological conditions such as cerebral vasospasm following subarachnoid hemorrhage and coronary arterial vasospasm. ILK and ZIPK may, therefore, be useful therapeutic targets for the treatment of such conditions.
Asunto(s)
Músculo Liso Vascular/fisiología , Miosina Tipo II/química , Miosina Tipo II/fisiología , Vasoconstricción/genética , Lesión Renal Aguda/tratamiento farmacológico , Animales , Catálisis , Vasoespasmo Coronario/tratamiento farmacológico , Proteínas Quinasas Asociadas a Muerte Celular/fisiología , Proteínas Quinasas Asociadas a Muerte Celular/uso terapéutico , Endotelina-1/farmacología , Humanos , Hipertensión/tratamiento farmacológico , Microcirculación/efectos de los fármacos , Microcirculación/genética , Terapia Molecular Dirigida , Quinasa de Cadena Ligera de Miosina/fisiología , Fosfatasa de Miosina de Cadena Ligera/fisiología , Fosforilación , Proteínas Serina-Treonina Quinasas/fisiología , Proteínas Serina-Treonina Quinasas/uso terapéutico , Ratas , Circulación Renal/efectos de los fármacos , Circulación Renal/genética , Vasoespasmo Intracraneal/tratamiento farmacológicoRESUMEN
Parathyroid hormone-related protein (PTHrP) belongs to vasoactive factors that regulate blood pressure and renal hemodynamics both by reducing vascular tone and raising renin release. PTHrP is expressed in systemic and renal vasculature. Here, we wanted to assess the contribution of vascular smooth muscle cell endogenous PTHrP to the regulation of cardiovascular and renal functions. We generated a mouse strain (SMA-CreERT2/PTHrPL2/L2 or premutant PTHrPSM-/-), which allows temporally controlled, smooth muscle-targeted PTHrP knockdown in adult mice. Tamoxifen treatment induced efficient recombination of PTHrP-floxed alleles and decreased PTHrP expression in vascular and visceral smooth muscle cells of PTHrPSM-/- mice. Blood pressure remained unchanged in PTHrPSM-/- mice, but plasma renin concentration and creatinine clearance were reduced. Renal hemodynamics were further analyzed during clearance measurements in anesthetized mice. Conditional knockdown of PTHrP decreased renal plasma flow and glomerular filtration rate with concomitant reduction in filtration fraction. Similar measurements were repeated during acute saline volume expansion. Saline volume expansion induced a rise in renal plasma flow and reduced filtration fraction; both were blunted in PTHrPSM-/- mice leading to impaired diuresis. These findings show that endogenous vascular smooth muscle PTHrP controls renal hemodynamics under basal conditions, and it is an essential factor in renal vasodilation elicited by saline volume expansion.
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Presión Sanguínea/genética , Presión Sanguínea/fisiología , Miocitos del Músculo Liso/metabolismo , Proteína Relacionada con la Hormona Paratiroidea/fisiología , Circulación Renal/genética , Circulación Renal/fisiología , Animales , Antineoplásicos Hormonales/farmacología , Volumen Sanguíneo/fisiología , Cartilla de ADN , Frecuencia Cardíaca/efectos de los fármacos , Frecuencia Cardíaca/fisiología , Inmunohistoquímica , Pruebas de Función Renal , Ratones , Ratones Noqueados , Ratones Transgénicos , Células Musculares/fisiología , Miocitos del Músculo Liso/efectos de los fármacos , Proteína Relacionada con la Hormona Paratiroidea/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Renina/metabolismo , Tamoxifeno/farmacologíaRESUMEN
Although the two-kidney, one-clip (2K1C) model is widely used as a model of human renovascular hypertension, mechanisms leading to the development of fibrosis and atrophy in the cuffed kidney and compensatory hyperplasia in the contralateral kidney have not been defined. Based on the well-established role of the transforming growth factor (TGF)-ß signaling pathway in renal fibrosis, we tested the hypothesis that abrogation of TGF-ß/Smad3 signaling would prevent fibrosis in the cuffed kidney. Renal artery stenosis (RAS) was established in mice with a targeted disruption of exon 2 of the Smad3 gene (Smad3 KO) and wild-type (WT) controls by placement of a polytetrafluoroethylene cuff on the right renal artery. Serial pulse-wave Doppler ultrasound assessments verified that blood flow through the cuffed renal artery was decreased to a similar extent in Smad3 KO and WT mice. Two weeks after surgery, systolic blood pressure and plasma renin activity were significantly elevated in both the Smad3 KO and WT mice. The cuffed kidney of WT mice developed renal atrophy (50% reduction in weight after 6 wk, P < 0.0001), which was associated with the development of interstitial fibrosis, tubular atrophy, and interstitial inflammation. Remarkably, despite a similar reduction of renal blood flow, the cuffed kidney of the Smad3 KO mice showed minimal atrophy (9% reduction in weight, P = not significant), with no significant histopathological alterations (interstitial fibrosis, tubular atrophy, and interstitial inflammation). We conclude that abrogation of TGF-ß/Smad3 signaling confers protection against the development of fibrosis and atrophy in RAS.
Asunto(s)
Hipertensión Renovascular/genética , Hipertensión Renovascular/patología , Riñón/patología , Proteína smad3/deficiencia , Proteína smad3/genética , Animales , Atrofia , Colágeno/biosíntesis , Constricción Patológica , Fibrosis , Inmunohistoquímica , Pruebas de Función Renal , Ratones , Mutación/genética , Mutación/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Obstrucción de la Arteria Renal/patología , Circulación Renal/genética , Circulación Renal/fisiología , Renina/sangre , Transducción de Señal/genética , Transducción de Señal/fisiología , Factor de Crecimiento Transformador beta/biosíntesisRESUMEN
NK cell transcripts are increased in biopsies with antibody-mediated rejection, whereas T cell transcripts are increased in T cell-mediated rejection. However, NK and T cells share many features, creating potential ambiguity. Therefore to estimate the NK- versus T cell transcript burdens separately, we defined nonoverlapping transcripts selective for NK cells (N = 4) or T cells (N = 5). We compared NK- versus T cell transcript burdens in microarrays from 403 kidney transplant biopsies (182 early, 221 late). In late biopsies, high NK-cell transcript expression was associated with antibody-mediated rejection, correlating with microvascular inflammation and donor specific HLA antibody. However, some early biopsies with T cell-mediated rejection had high NK-cell transcript expression, as well as T cell transcripts, without evidence of antibody-mediated rejection or DSA, correlating with interstitial inflammation and tubulitis. Both NK-cell and T cell transcripts were moderately increased in many kidneys with inflammation secondary to injury or atrophy scarring. These results support the distinct role of NK cells in late antibody-mediated rejection, but indicate a role for NK-transcript expressing cells (NK cells or T cells with NK features) both in T cell-mediated rejection and in inflammation associated with injury and atrophy scarring.
Asunto(s)
Biomarcadores/metabolismo , Perfilación de la Expresión Génica , Rechazo de Injerto/genética , Trasplante de Riñón/inmunología , Células Asesinas Naturales/metabolismo , Linfocitos T/metabolismo , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Estudios de Seguimiento , Rechazo de Injerto/inmunología , Supervivencia de Injerto/genética , Supervivencia de Injerto/inmunología , Humanos , Inflamación/genética , Inflamación/inmunología , Isoanticuerpos/sangre , Isoanticuerpos/inmunología , Células Asesinas Naturales/inmunología , Masculino , Microcirculación/genética , Microcirculación/inmunología , Persona de Mediana Edad , Análisis de Secuencia por Matrices de Oligonucleótidos , ARN Mensajero/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Circulación Renal/genética , Circulación Renal/inmunología , Linfocitos T/inmunología , Adulto JovenRESUMEN
Previous studies demonstrate a role for ß epithelial Na(+) channel (ßENaC) protein as a mediator of myogenic constriction in renal interlobar arteries. However, the importance of ßENaC as a mediator of myogenic constriction in renal afferent arterioles, the primary site of development of renal vascular resistance, has not been determined. We colocalized ßENaC with smooth muscle α-actin in vascular smooth muscle cells in renal arterioles using immunofluorescence. To determine the importance of ßENaC in myogenic constriction in renal afferent arterioles, we used a mouse model of reduced ßENaC (ßENaC m/m) and examined pressure-induced constrictor responses in the isolated afferent arteriole-attached glomerulus preparation. We found that, in response to a step increase in perfusion pressure from 60 to 120 mmHg, the myogenic tone increased from 4.5 ± 3.7 to 27.3 ± 5.2% in +/+ mice. In contrast, myogenic tone failed to increase with the pressure step in m/m mice (3.9 ± 0.8 to 6.9 ± 1.4%). To determine the importance of ßENaC in myogenic renal blood flow (RBF) regulation, we examined the rate of change in renal vascular resistance following a step increase in perfusion pressure in volume-expanded animals. We found that, following a step increase in pressure, the rate of myogenic correction of RBF is inhibited by 75% in ßENaC m/m mice. These findings demonstrate that myogenic constriction in afferent arterioles is dependent on normal expression of ßENaC.
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Arteriolas/fisiología , Canales Epiteliales de Sodio/fisiología , Músculo Liso Vascular/fisiología , Circulación Renal/fisiología , Actinas/metabolismo , Actinas/fisiología , Animales , Animales Modificados Genéticamente , Presión Sanguínea/genética , Presión Sanguínea/fisiología , Interpretación Estadística de Datos , Canales Epiteliales de Sodio/genética , Glomérulos Renales/irrigación sanguínea , Glomérulos Renales/metabolismo , Ratones , Tono Muscular/genética , Tono Muscular/fisiología , Circulación Renal/genética , Resistencia Vascular/genética , Resistencia Vascular/fisiología , Vasoconstricción/genética , Vasoconstricción/fisiologíaRESUMEN
BACKGROUND/AIMS: Renal interstitial fibrosis is a final common pathway of all chronic, progressive kidney diseases. Peritubular capillary rarefaction is strongly correlated with fibrosis. The adherens junction protein vascular endothelial cadherin (VE-cadherin) is thought to play a critical role in vascular integrity. We hypothesized that VE-cadherin modulates the renal microcirculation during fibrogenesis and ultimately affects renal fibrosis. METHODS: Unilateral ureteral obstruction (UUO) was used as a renal fibrosis model in VE-cadherin heterozygote (VE+/-) and wild-type (WT) mice, and the kidneys were harvested at days 3, 7, and 14. Peritubular capillary changes and fibrogenesis were investigated. RESULTS: VE+/- mice had lower levels of VE-cadherin protein than WT mice at 3 and 7, but not 14 days after UUO. Vascular permeability was significantly greater in VE+/- mice 7 days after UUO, while peritubular capillary density was not significantly different in VE+/- and WT mice. Interstitial myofibroblast numbers and collagen I and III mRNA levels were significantly higher in VE+/- mice, consistent with a stronger early fibrogenic response. Expression of the pericyte marker neuron-glial antigen 2 was upregulated after UUO, but was not greater in VE+/- mice compared to the WT mice. CONCLUSION: Our data suggest that VE-cadherin controls vascular permeability and limits fibrogenesis after UUO.
Asunto(s)
Antígenos CD/metabolismo , Cadherinas/metabolismo , Modelos Animales de Enfermedad , Riñón/metabolismo , Obstrucción Ureteral/metabolismo , Animales , Antígenos/genética , Antígenos/metabolismo , Antígenos CD/genética , Antígenos de Diferenciación Mielomonocítica/metabolismo , Western Blotting , Cadherinas/genética , Permeabilidad Capilar/genética , Permeabilidad Capilar/fisiología , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Colágeno Tipo III/genética , Colágeno Tipo III/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , Fibrosis , Expresión Génica , Heterocigoto , Inmunohistoquímica , Riñón/irrigación sanguínea , Riñón/patología , Macrófagos/metabolismo , Macrófagos/patología , Ratones , Ratones Noqueados , Miofibroblastos/metabolismo , Miofibroblastos/patología , Proteoglicanos/genética , Proteoglicanos/metabolismo , Circulación Renal/genética , Circulación Renal/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Obstrucción Ureteral/genética , Obstrucción Ureteral/fisiopatologíaRESUMEN
OBJECTIVE AND METHODS: A loss-of-function cytosine (C) for thymidine (T) transition at nucleotide 8590 of CYP4A11 has been associated with increased blood pressure in humans. We tested the hypothesis that CYP4A11 T8590C genotype is associated with salt sensitivity in the International Hypertensive Pathotype cohort. RESULTS: CYP4A11 T8590C genotype was associated with hypertension in whites. Among normotensive individuals, CYP4A11 T8590C genotype was associated with mean arterial pressure (MAP) during both high and low salt diets, such that there was no relationship between genotype and salt sensitivity of blood pressure. Among hypertensive individuals, CYP4A11 T8590C genotype did not associate with MAP during high salt intake, whereas MAP decreased with increasing number of C alleles during salt restriction. Consequently, among hypertensive individuals, change in MAP with salt restriction was greatest in individuals homozygous for the C allele (-10.9â±â9.9, -11.1â±â12.3, and -18.8â±â12.0âmmHg in TT, CT, and CC groups, respectively, Pâ=â0.02). In both normotensive and hypertensive individuals, individuals homozygous for the C allele exhibited an attenuated increase in renal blood flow during high salt. CYP4A11 genotype did not affect pressor responses to Angiotensin II in normotensive or hypertensive individuals. CONCLUSION: The loss-of-function CYP4A11 8590C allele is associated with a diagnosis of hypertension and, in normotensive individuals, with higher blood pressure regardless of salt intake. Among hypertensive individuals, the C allele is associated with salt-sensitive blood pressure. Impaired renal vasodilation during high salt intake may contribute to salt sensitivity. Studies are needed to determine whether CYP4A11 T8590C genotype predicts responses to medications that affect sodium homeostasis in hypertensive individuals.
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Sistema Enzimático del Citocromo P-450/genética , Hipertensión/genética , Circulación Renal/genética , Adulto , Alelos , Angiotensina II/farmacología , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/genética , Presión Sanguínea/fisiología , Citocromo P-450 CYP4A , Dieta Hiposódica , Femenino , Estudios de Asociación Genética , Humanos , Hipertensión/dietoterapia , Hipertensión/etiología , Hipertensión/fisiopatología , Masculino , Persona de Mediana Edad , Norepinefrina/farmacología , Polimorfismo de Nucleótido Simple , Circulación Renal/fisiología , Cloruro de Sodio Dietético/administración & dosificaciónRESUMEN
The Notch3 receptor participates in the development and maturation of vessels. Mutations of Notch3 in humans are associated with defective regulation of cerebral blood flow. To investigate the role of Notch3 in the regulation of renal hemodynamics, we used mice lacking expression of the Notch3 gene (Notch3-/- mice). Bolus injections of norepinephrine and angiotensin II increased renal vascular resistance and decreased renal blood flow in a dose-dependent manner in wild-type mice. In sharp contrast, renal vascular resistance of Notch3-/- mice varied little after boluses of norepinephrine and angiotensin II. Inversely, bradykinin and prostacyclin relaxed renal vasculature in wild-type mice. Both vasodilators had a negligible effect on renal vascular resistance of Notch3-/- mice. Afferent arterioles freshly isolated from Notch3-/- mice displayed decreased thickness of vascular wall compared with wild -type mice and showed a deficient contractile response to angiotensin II. To examine the physiopathological consequences of the above-described deficiency, hypertension was induced by continuous infusion of angiotensin II. Angiotensin II gradually increased blood pressure in both strains, but this increase was lesser in the Notch3-/- mice. Despite this blunted systemic effect, Notch3-/- mice displayed high mortality rates (65%) attributed to heart failure. In the kidney, the surviving Notch3-/- mice showed focal structural alterations characteristic of nephroangiosclerosis. These data show that Notch3 is necessary for the adaptive response of the renal vasculature to vasoactive systems. A deficiency in the expression of Notch3 could have important physiopathological consequences in the adaptation of the cardiac and renal function to chronic increase of blood pressure.
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Riñón/irrigación sanguínea , Receptores Notch/fisiología , Circulación Renal/fisiología , Resistencia Vascular/fisiología , Angiotensina II/farmacología , Animales , Arteriolas/efectos de los fármacos , Arteriolas/fisiología , Presión Sanguínea/genética , Presión Sanguínea/fisiología , Bradiquinina/farmacología , Epoprostenol/farmacología , Expresión Génica , Humanos , Hipertensión/inducido químicamente , Hipertensión/genética , Hipertensión/fisiopatología , Inmunohistoquímica , Técnicas In Vitro , Riñón/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Norepinefrina/farmacología , Receptor Notch3 , Receptores Notch/genética , Receptores Notch/metabolismo , Circulación Renal/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Piel/irrigación sanguínea , Resistencia Vascular/genética , Vasoconstrictores/farmacología , Vasodilatadores/farmacologíaRESUMEN
The present study examined the effect of transfer of portions of chromosome 1 that includes (FHH.1(BN) AR(+) strain) or excludes (control FHH.1(BN) AR(-) strain) a 4.3-Mb region from the Brown Norway (BN) rat that restores the autoregulation (AR) of renal blood flow (RBF) on the development of hypertension and renal injury in congenic strains of Fawn Hooded Hypertensive (FHH) rats. FHH and control AR(-) rats exhibited poor autoregulation of RBF, and glomerular capillary pressure (Pgc) rose by 19 ± 2 mmHg in FHH rats when renal perfusion pressure (RPP) was increased from 100 to 150 mmHg. In contrast, RBF was well autoregulated in the AR(+) strain, and Pgc only increased by 3 ± 1 mmHg when RPP was increased over this range. Baseline mean arterial pressure (MAP) at 12 wk of age was similar in all strains and averaged 122 mmHg. MAP increased significantly in FHH rats and was significantly higher by 12 mmHg in 21-wk-old FHH rats than in the FHH.1(BN) congenic strains. Protein excretion rose from 5 ± 1 to 397 ± 29 mg/day in 6- vs. 21-wk-old FHH rats. In contrast, protein excretion only increased to 139 ± 21 mg/day in the control AR(-) strain, and it did not increase significantly in the AR(+) strain. Glomerular permeability to albumin was similar in all strains at 6 wk of age. It increased significantly in 9-wk-old FHH and control AR(-) rats, but not in the AR(+) strain. The levels of matrix metalloproteinase (MMP)-2 and transforming growth factor (TGF)-ß2 protein were significantly higher in the renal cortex of 9-wk-old FHH rats compared with the levels seen in the AR(+) strain. These data indicate that transfer of a 4.3-Mb region of BN chromosome 1 into the FHH genetic background improves autoregulation of RBF, normalizes Pgc, and slows the progression of renal disease.
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Hipertensión Renal/genética , Enfermedades Renales/genética , Proteinuria/genética , Circulación Renal/genética , Animales , Cromosomas de los Mamíferos/genética , Técnicas de Transferencia de Gen , Tasa de Filtración Glomerular/genética , Homeostasis/genética , Hipertensión Renal/fisiopatología , Riñón/irrigación sanguínea , Riñón/fisiopatología , Glomérulos Renales/fisiopatología , Masculino , Metaloproteinasa 2 de la Matriz/análisis , Ratas , Ratas Endogámicas BN , Factor de Crecimiento Transformador beta2/análisisRESUMEN
The production of 20-hydroxyeicosatetraenoic acid (20-HETE) in the kidney is thought to be involved in the control of renal vascular tone and tubular sodium and chloride reabsorption. Cytochrome (Cyp) P-450 enzymes of the Cyp4a family in the mouse, namely 4a10, -12 and 14, are involved in 20-HETE synthesis. Recent advances in the molecular genetics of the mouse have produced mice in which Cyp4a isoforms have been disrupted and the consequence of such an approach is examined. This study evaluated the effect of deletion of the Cyp4a14 gene on blood pressure, renal vascular responses and tubular function. When compared with the wild-type (WT) litter mates, systolic blood pressure was greater in Cyp4a14 null (KO) mice as were renal vascular responses to angiotensin II or phenyephrine, G protein-coupled receptor (GPCR) agonists, but not KCl, a non-GPCR agonist. Renal vascular responses to guanosine 5'-O-(gamma-thio)triphosphate, a non-hydrolyzable GTP analog, or NaF(4), an activator of G-proteins, were also enhanced. However, vasodilation to bradykinin or apocynin but not sodium nitroprusside was blunted in Cyp4a14 null (KO) kidneys. These changes in KO mice were accompanied by increased 20-HETE synthesis, reduced renal production of nitric oxide (NO), increased lipid hydroperoxides and increased apocynin-inhibitable vascular NADPH oxidase activity that was prevented by administration of NO synthase (NOS) inhibitor, suggesting endothelial nitric oxide synthase (eNOS) uncoupling. Cyp4a14 KO mice also exhibited a diminished capacity to excrete an acute sodium load (0.9% NaCl, 2.5 mL/kg). These data suggest that deletion of the Cyp4a gene conferred a prohypertensive status via mechanisms involving increased 20-HETE synthesis and eNOS uncoupling leading to increased oxidative stress, enhanced vasoconstriction but diminished vasodilation as well as a defect in the renal excretory capacity in Cyp4a14 KO mice. These mechanisms suggest that the Cyp4a14-deficient mouse may be a useful model for evaluation of NO/20-HETE interactions.
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Presión Sanguínea/genética , Sistema Enzimático del Citocromo P-450/genética , Riñón/fisiología , Circulación Renal/genética , Animales , Sistema Enzimático del Citocromo P-450/fisiología , Familia 4 del Citocromo P450 , Inhibidores Enzimáticos/farmacología , Femenino , Eliminación de Gen , Ácidos Hidroxieicosatetraenoicos/biosíntesis , Ácidos Hidroxieicosatetraenoicos/genética , Ácidos Hidroxieicosatetraenoicos/fisiología , Hipertensión/genética , Técnicas In Vitro , Riñón/metabolismo , Peróxidos Lipídicos/metabolismo , Masculino , Ratones , Ratones Noqueados , Microsomas/metabolismo , NADPH Oxidasas/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa/antagonistas & inhibidores , Nitritos/orina , Cloruro de Sodio/metabolismo , Vasoconstricción/genéticaRESUMEN
Calcineurin inhibitors (CNI) are powerful immunomodulatory agents that produce marked renal dysfunction due in part to endothelin-1-mediated reductions in renal blood flow. Ligand-stimulated Gq protein signaling promotes the contraction of smooth muscle cells via phospholipase Cbeta-mediated stimulation of cytosolic calcium release. RGS4 is a GTPase activating protein that promotes the deactivation of Gq and Gi family members. To investigate the role of G protein-mediated signaling in the pathogenesis of CNI-mediated renal injury, we used mice deficient for RGS4 (rgs4(-/-)). Compared to congenic wild type control animals, rgs4(-/-) mice were intolerant of the CNI, cyclosporine (CyA), rapidly developing fatal renal failure. Rgs4(-/-) mice exhibited markedly reduced renal blood flow after CyA treatment when compared to congenic wild type control mice as measured by magnetic resonance imaging (MRI). Hypoperfusion was reversed by coadministration of CyA with the endothelin antagonist, bosentan. The MAPK/ERK pathway was activated by cyclosporine administration and was inhibited by cotreatment with bosentan. These results show that endothelin-1-mediated Gq protein signaling plays a key role in the pathogenesis of vasoconstrictive renal injury and that RGS4 antagonizes the deleterious effects of excess endothelin receptor activation in the kidney.
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Proteínas de Unión al GTP/metabolismo , Proteínas/metabolismo , Animales , Calcio/metabolismo , Ciclosporina/metabolismo , Endotelina-1/genética , Endotelina-1/metabolismo , Subunidades alfa de la Proteína de Unión al GTP Gq-G11/genética , Subunidades alfa de la Proteína de Unión al GTP Gq-G11/metabolismo , Proteínas de Unión al GTP/genética , Proteínas Activadoras de GTPasa/genética , Proteínas Activadoras de GTPasa/metabolismo , Ratones , Ratones Congénicos , Ratones Noqueados , Miocitos del Músculo Liso/metabolismo , Proteínas del Tejido Nervioso , Proteínas/genética , Circulación Renal/genética , Transducción de Señal/genética , Transducción de Señal/fisiologíaRESUMEN
1. Renal medullary blood flow (MBF) is believed today to have a potent role in blood pressure control through its influence on sodium and water excretion. The present article reviews: (i) the study of MBF in two experimental models of heritable hypertension, namely spontaneously hypertensive rats (SHR) and Lyon genetically hypertensive (LH) rats selected, respectively, from Wistar and Sprague-Dawley rats; and (ii) the regulation of MBF by the renin-angiotensin system (RAS) in these animals and the interaction between angiotensin (Ang) II and nitric oxide, prostaglandins and reactive oxygen species in the renal medulla. 2. Although numerous renal disorders are observed during or after the development of hypertension, the reduced pressure-natriuresis function is an early apparent and common abnormality found in SHR and LH rats. This abnormality is associated with a blunted increase in MBF in response to elevations in renal perfusion pressure. Moreover, both SHR and LH rats exhibit an exaggerated medullary vasoconstriction and/or a reduced medullary vasodilatation under stimulation with AngII. 3. Chronic RAS blockade prevents the development of hypertension in both SHR and LH rats. This effect involves an improved response of MBF to increasing renal perfusion pressure and to AngII. 4. Medullary blood flow is inappropriately regulated in genetically hypertensive rats; a decreased MBF caused by the disequilibrium of local vasodilator/vasoconstrictor systems may favour the prohypertensive role of the RAS in genetic hypertension.
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Hipertensión/genética , Hipertensión/fisiopatología , Médula Renal/irrigación sanguínea , Sistema Renina-Angiotensina/fisiología , Animales , Presión Sanguínea/fisiología , Hipertensión/patología , Ratas , Ratas Endogámicas SHR , Ratas Sprague-Dawley , Ratas Wistar , Circulación Renal/genética , Circulación Renal/fisiologíaRESUMEN
Ischemia followed by reperfusion leads to severe organ injury and dysfunction. Inflammation is considered to be the most important cause of graft dysfunction in kidney transplantation subjected to ischemia. The mechanism that triggers inflammation and renal injury after ischemia remains to be elucidated; however, cellular stress may induce apoptosis during the first hours and days after transplantation, which might play a crucial role in early graft dysfunction. Bcl-2 is known to inhibit apoptosis induced by the etiological factors promoting ischemia and reperfusion injury. Accordingly, we hypothesized that an augmentation of the antiapoptotic factor Bcl-2 may thus protect tubular epithelial cells by inhibiting apoptosis, thereby ameliorating the subsequent tubulointerstitial injury. We examined the effects of Bcl-2 overexpression on ischemia-reperfusion (I/R) injury using Bcl-2 transgenic mice (Bcl-2 TG) and their wild-type littermates (WT). To investigate the effects of I/R injury, the left renal artery and vein were clamped for 45 min, followed by reperfusion for 0-96 h. Bcl-2 TG exhibited decreased active caspase protein in the tubular cells, which led to a reduction in TUNEL-positive apoptotic cells. Consequently, interstitial fibrosis and phenotypic changes were ameliorated in Bcl-2 TG. In conclusion, Bcl-2 augmentation protected renal tubular epithelial cells from I/R, and subsequent interstitial injury by inhibiting tubular apoptosis.