RESUMEN
BACKGROUND: Kidney injury associated with cold storage is a determinant of delayed graft function and the long-term outcome of transplanted kidneys, but the underlying mechanism remains elusive. We previously reported a role of protein kinase C-δ (PKCδ) in renal tubular injury during cisplatin nephrotoxicity and albumin-associated kidney injury, but whether PKCδ is involved in ischemic or transplantation-associated kidney injury is unknown. METHODS: To investigate PKCδ's potential role in injury during cold storage-associated transplantation, we incubated rat kidney proximal tubule cells in University of Wisconsin (UW) solution at 4°C for cold storage, returning them to normal culture medium at 37°C for rewarming. We also stored kidneys from donor mice in cold UW solution for various durations, followed by transplantation into syngeneic recipient mice. RESULTS: We observed PKCδ activation in both in vitro and in vivo models of cold-storage rewarming or transplantation. In the mouse model, PKCδ was activated and accumulated in mitochondria, where it mediated phosphorylation of a mitochondrial fission protein, dynamin-related protein 1 (Drp1), at serine 616. Drp1 activation resulted in mitochondrial fission or fragmentation, accompanied by mitochondrial damage and tubular cell death. Deficiency of PKCδ in donor kidney ameliorated Drp1 phosphorylation, mitochondrial damage, tubular cell death, and kidney injury during cold storage-associated transplantation. PKCδ deficiency also improved the repair and function of the renal graft as a life-supporting kidney. An inhibitor of PKCδ, δV1-1, protected kidneys against cold storage-associated transplantation injury. CONCLUSIONS: These results indicate that PKCδ is a key mediator of mitochondrial damage and renal tubular injury in cold storage-associated transplantation and may be an effective therapeutic target for improving renal transplant outcomes.
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Frío/efectos adversos , Dinaminas/metabolismo , Trasplante de Riñón , Necrosis Tubular Aguda/etiología , Túbulos Renales Proximales/enzimología , Preservación de Órganos/métodos , Proteína Quinasa C-delta/fisiología , Animales , Apoptosis , División Celular , Células Cultivadas , Activación Enzimática , Necrosis Tubular Aguda/enzimología , Túbulos Renales Proximales/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Mitocondrias/enzimología , Fosforilación , Proteína Quinasa C-delta/antagonistas & inhibidores , Inhibidores de Proteínas Quinasas/farmacología , Procesamiento Proteico-Postraduccional , RatasRESUMEN
Acute kidney injury (AKI) causes severe morbidity, mortality, and chronic kidney disease (CKD). Mortality is particularly marked in the elderly and with preexisting CKD. Oxidative stress is a common theme in models of AKI induced by ischemia-reperfusion (I-R) injury. We recently characterized an intracellular isoform of matrix metalloproteinase-2 (MMP-2) induced by oxidative stress-mediated activation of an alternate promoter in the first intron of the MMP-2 gene. This generates an NH2-terminal truncated MMP-2 (NTT-MMP-2) isoform that is intracellular and associated with mitochondria. The NTT-MMP-2 isoform is expressed in kidneys of 14-mo-old mice and in a mouse model of coronary atherosclerosis and heart failure with CKD. We recently determined that NTT-MMP-2 is induced in human renal transplants with delayed graft function and correlated with tubular cell necrosis. To determine mechanism(s) of action, we generated proximal tubule cell-specific NTT-MMP-2 transgenic mice. Although morphologically normal at the light microscopic level at 4 mo, ultrastructural studies revealed foci of tubular epithelial cell necrosis, the mitochondrial permeability transition, and mitophagy. To determine whether NTT-MMP-2 expression enhances sensitivity to I-R injury, we performed unilateral I-R to induce mild tubular injury in wild-type mice. In contrast, expression of the NTT-MMP-2 isoform resulted in a dramatic increase in tubular cell necrosis, inflammation, and fibrosis. NTT-MMP-2 mice had enhanced expression of innate immunity genes and release of danger-associated molecular pattern molecules. We conclude that NTT-MMP-2 "primes" the kidney to enhanced susceptibility to I-R injury via induction of mitochondrial dysfunction. NTT-MMP-2 may be a novel AKI treatment target.
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Lesión Renal Aguda/enzimología , Necrosis Tubular Aguda/enzimología , Túbulos Renales Proximales/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Daño por Reperfusión/enzimología , Lesión Renal Aguda/genética , Lesión Renal Aguda/inmunología , Lesión Renal Aguda/patología , Factores de Edad , Animales , Enfermedad de la Arteria Coronaria/enzimología , Enfermedad de la Arteria Coronaria/genética , Enfermedad de la Arteria Coronaria/patología , Modelos Animales de Enfermedad , Predisposición Genética a la Enfermedad , Insuficiencia Cardíaca/enzimología , Insuficiencia Cardíaca/genética , Insuficiencia Cardíaca/patología , Humanos , Inmunidad Innata , Isoenzimas , Necrosis Tubular Aguda/genética , Necrosis Tubular Aguda/inmunología , Necrosis Tubular Aguda/patología , Túbulos Renales Proximales/inmunología , Túbulos Renales Proximales/ultraestructura , Metaloproteinasa 2 de la Matriz/genética , Potencial de la Membrana Mitocondrial , Ratones Noqueados , Ratones Transgénicos , Mitocondrias/enzimología , Mitocondrias/ultraestructura , Mitofagia , Infarto del Miocardio/enzimología , Infarto del Miocardio/genética , Infarto del Miocardio/patología , Necrosis , Estrés Oxidativo , Fenotipo , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/genética , Daño por Reperfusión/inmunología , Daño por Reperfusión/patología , Transducción de SeñalRESUMEN
BACKGROUND: Serum lactate dehydrogenase (LDH) levels may help to distinguish ischemic acute tubular necrosis (ATN) from acute rejection after kidney transplantation. METHODS: All kidney biopsies performed in the years 2010 to 2012 were reviewed. Serum LDH, creatinine level, clinical variables, and presence of donor-specific antibodies were recorded before the biopsy. RESULTS: Overall 150 biopsies were included. Ischemic ATN was diagnosed in 45 biopsies and acute cellular-mediated rejection and/or antibody-mediated rejection in 59 biopsies, 38 of which were accompanied by ATN. Serum LDH was elevated in 23 (51%) of 45 cases with ischemic ATN versus 15 (14%) of 105 cases with other diagnoses ( P < .0001). Median serum LDH was 478 U/L (range 277-2018) for ischemic ATN and 372 U/L (range 191-748) for all other diagnoses ( P < .001). When delayed graft function or primary nonfunctioning grafts were caused by ischemic ATN, serum LDH was elevated in 58% of cases, but when caused by acute rejection, LDH was normal in 88% of cases ( P = .02). CONCLUSIONS: There is a strong association between elevated serum LDH 1 to 3 days before performing kidney biopsy and the diagnosis of ischemic ATN after kidney transplantation, especially at the immediate posttransplantation period. Normal serum LDH at this period should raise a suspicion of acute rejection.
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Rechazo de Injerto/enzimología , Trasplante de Riñón , Necrosis Tubular Aguda/enzimología , Lactato Deshidrogenasas/sangre , Adulto , Biomarcadores/sangre , Biopsia , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Nephrotoxicity limits the therapeutic efficacy of the antineoplastic drug cisplatin. Due to dosage adjustment and appropriate monitoring, most therapeutic courses with cisplatin produce no or minimal kidney damage. However, we studied whether even sub-nephrotoxic dosage of cisplatin poses a potential risk for the kidneys by predisposing to acute kidney injury (AKI), specifically by lowering the toxicity threshold for a second nephrotoxin. With this purpose rats were treated with a single sub-nephrotoxic dosage of cisplatin (3mg/kg, i.p.) and after two days, with a sub-nephrotoxic regime of gentamicin (50mg/kg/day, during 6 days, i.p.). Control groups received only one of the drugs or the vehicle. Renal function and renal histology were monitored throughout the experiment. Cisplatin treatment did not cause any relevant functional or histological alterations in the kidneys. Rats treated with cisplatin and gentamicin, but not those under single treatments, developed an overt renal failure characterized by both renal dysfunction and massive tubular necrosis. In addition, the urinary excretion of fumarylacetoacetase was increased in cisplatin-treated animals at subtoxic doses, which might be exploited as a cisplatin-induced predisposition marker. In fact, the urinary level of fumarylacetoacetase prior to the second nephrotoxin correlated with the level of AKI triggered by gentamicin in predisposed animals.
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Lesión Renal Aguda/inducido químicamente , Antineoplásicos/toxicidad , Cisplatino/toxicidad , Hidrolasas/orina , Riñón/efectos de los fármacos , Lesión Renal Aguda/enzimología , Lesión Renal Aguda/patología , Lesión Renal Aguda/fisiopatología , Lesión Renal Aguda/orina , Animales , Biomarcadores/orina , Modelos Animales de Enfermedad , Gentamicinas , Humanos , Riñón/enzimología , Riñón/patología , Riñón/fisiopatología , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/orina , Masculino , Ratas Wistar , Medición de Riesgo , Factores de Riesgo , Factores de Tiempo , Regulación hacia ArribaRESUMEN
BACKGROUND: Matrix metalloproteinase-9 (MMP-9) is the most important member of the MMP family responsible for the development and progression of various renal diseases. Our study aims to investigate the localization of MMP-9 in human renal allografts and to assess whether MMP-9 immunostaining is contributory to detect pathological change in renal biopsy. METHODS: We examined 150 renal allograft biopsies (48 baseline and 102 follow-up) from 49 transplants and analyzed the associations of clinical and histopathological data with the MMP-9 staining intensity using a semi-quantitative scoring. RESULTS: MMP-9 immunostaining in proximal tubule epithelium was negative before transplantation, but positive in biopsies with episodes, particularly with acute cellular rejection (ACR) and acute calcineurin inhibitor (CNI) toxicity. Tubulitis was the most significant association factor (p < 0.0001) with increased MMP-9 staining intensity. The expression in proximal tubules remained augmented in allografts recovered from ACR episodes, while it was disappeared or diminished in those recovered from acute CNI toxicity or ischemia/reperfusion effects. CONCLUSION: These findings suggest the necessary participation of MMP-9 in the pathogenesis of tubulitis and the subsequent stage of pathogenesis in ACR. Up-regulation of MMP-9 expression in the proximal tubule could be a new indicator of tubular injury and a predictive factor for the prognosis of renal allograft.
Asunto(s)
Rechazo de Injerto/enzimología , Rechazo de Injerto/prevención & control , Trasplante de Riñón , Necrosis Tubular Aguda/enzimología , Metaloproteinasa 9 de la Matriz/inmunología , Metaloproteinasa 9 de la Matriz/metabolismo , Adolescente , Adulto , Niño , Terapia Combinada , Femenino , Humanos , Técnicas para Inmunoenzimas , Inmunosupresores/uso terapéutico , Necrosis Tubular Aguda/terapia , Masculino , Persona de Mediana Edad , Pronóstico , Trasplante Homólogo , Adulto JovenRESUMEN
Ischemia-reperfusion injury (IRI) is a leading cause of acute tubular necrosis (ATN) and delayed graft function in transplanted organs. Up-regulation of matrix metalloproteinases (MMPs) propagates the microinflammatory response that drives IRI. This study sought to determine the specific effects of Marimastat (Vernalis, BB-2516), a broad spectrum MMP and TNF-alpha-converting enzyme inhibitor, on IRI-induced ATN. Mice were pretreated with Marimastat or methylcellulose vehicle for 4 d before surgery. Renal pedicles were bilaterally occluded for 30 min and allowed to reperfuse for 24 h. Baseline creatinine levels were consistent between experimental groups; however, post-IRI creatinine levels were 4-fold higher in control mice (p < 0.0001). The mean difference between the post-IRI histology grades of Marimastat-treated and control kidneys was 1.57 (p = 0.003), demonstrating more severe damage to control kidneys. Post-IRI mean (+/-SEM) MMP-2 activity rose from baseline levels in control mice (3.62 +/- 0.99); however, pretreated mice presented only a slight increase in mean MMP-2 activity (1.57 +/- 0.72) (p < 0.001). In conclusion, these data demonstrate that MMP inhibition is associated with a reduction of IRI in a murine model.
Asunto(s)
Ácidos Hidroxámicos/farmacología , Necrosis Tubular Aguda/prevención & control , Riñón/efectos de los fármacos , Inhibidores de la Metaloproteinasa de la Matriz , Inhibidores de Proteasas/farmacología , Daño por Reperfusión/prevención & control , Animales , Biomarcadores/sangre , Western Blotting , Creatinina/sangre , Modelos Animales de Enfermedad , Riñón/irrigación sanguínea , Riñón/enzimología , Riñón/patología , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/patología , Masculino , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Ratones Endogámicos C57BL , Daño por Reperfusión/enzimología , Daño por Reperfusión/patología , Índice de Severidad de la EnfermedadRESUMEN
Nephropathic cystinosis is characterized clinically by generalized proximal renal tubular dysfunction, renal Fanconi Syndrome and progressive renal failure. Glomerular-proximal tubule disconnection has been noted in renal biopsies from patients with nephropathic cystinosis. In vitro studies performed in cystinotic fibroblasts and renal proximal tubular cells support a role for apoptosis of the glomerulotubular junction, and we have further extended these studies to human native cystinotic kidney specimens. We performed semi-quantitative analysis of tubular density in kidney biopsies from patients with nephropathic cystinosis and demonstrated a significant reduction (p=0.0003) in the number of proximal tubules in the kidney tissue of patients with cystinosis compared to normal kidneys and kidneys with other causes of renal injury; this reduction appears to be associated with the over-expression of caspase-4. This study provides the first quantitative evidence of a loss of proximal tubules in nephropathic cystinosis and suggests a possible role of caspase-4 in the apoptotic loss of proximal tubular cells. Further work is needed to elucidate if this injury mechanism may be causative for the progression of renal functional decline in nephropathic cystinosis.
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Lesión Renal Aguda/patología , Caspasas Iniciadoras/metabolismo , Necrosis Tubular Aguda/patología , Túbulos Renales Proximales/patología , Lesión Renal Aguda/metabolismo , Apoptosis , Niño , Cistinosis , Síndrome de Fanconi , Glomeruloesclerosis Focal y Segmentaria/enzimología , Glomeruloesclerosis Focal y Segmentaria/patología , Granulomatosis con Poliangitis/enzimología , Granulomatosis con Poliangitis/patología , Humanos , Necrosis Tubular Aguda/enzimología , Túbulos Renales Proximales/enzimología , Síndrome Nefrótico/enzimología , Síndrome Nefrótico/patología , Reflujo Vesicoureteral/enzimología , Reflujo Vesicoureteral/patologíaRESUMEN
Poli (ADP-Ribosa) polimerasa (PARP-1) cataliza la ADP ribosilación de proteínas usando NAD(+) como sustrato. La activación de PARP-1 conduce a la depleción intracelular de NAD(+). El daño por isquemia-reperfusión (IR) induce una activación excesiva de PARP-1 y la muerte celular por consumo masivo de ATP. Nuestra hipótesis de trabajo es que la excesiva expresión tubular de PARP-1 en riñones humanos trasplantados es una de las causas directas de la inducción de necrosis tubular aguda (NTA) y contribuye al retraso de la función renal del injerto. Material y Métodos: Estudiamos 193 biopsias de trasplante renal (95 preimplante biopsia de donante y 98 postrasplante) incluidas en parafina con diferentes grados de NTA y 65 biopsias renales de donante sin NTA. La NTA se estratificó en cuatro grados: ausente (0); leve (1) [50%]. La expresión nuclear de PARP-1 fue evaluada mediante inmunohistoquímica con el kit de polimeros conjugado con peroxidasa MasVision y el anticuerpo anti- PARP-1 (clón PARP01) y valorada semicuantitativamente de 0 a 3. Resultados: La expresión nuclear de PARP-1 antecedió a los cambios morfológicos sugerentes de NTA. Principalmente la inmunotinción se localizó en los núcleos de células tubulares, cuando fue intensa la lesión también apareció en glomérulos (epitelio de la cápsula de Bowman y células endoteliales de capilares glomerulares). La inmunotición fue observable hasta fases finales de la necrosis tubular. La totalidad de las 95 biopsias renales pre-trasplante con NTA grado 1 (86%) o grado 2 (14%) mostraron expresión nuclear de PARP-1 en túbulos. Las 98 biopsias postrasplante con NTA mostraron expresión más intensa de PARP-1 [grado 2 (45%), grado 3 (25%)]. El grado de NTA se correlacionó significantivamente con la expresión de PARP- 1(r=0,565, p=0,0001, test de Pearson), con una expresión media 2,74±0,45 en los casos de NTA severa frente a 1,94±0,74 en los casos de NTA leve y 0,29±0,45 en los casos sin NTA (p=0,0001, test ANOVA de una vía). En conclusión, PARP-1 está vinculado a la inducción de NTA y desempeña un papel importante en el comportamiento de la función precoz del injerto renal ya que está correlacionada significativamente con el tiempo en recuperar la diuresis eficaz (r=0,578, p=0,0001, test de Pearson) y con los niveles séricos de creatinina en el momento de la biopsia (r=0,649), y a los 3 meses (r=0,363, p=0,0001, test de Pearson) pero no a los 6 y 12 meses postrasplante
Poly (ADP-Ribose) Polymerase (PARP-1) catalyzes ADP-ribosylation of proteins using NAD(+) as substrate. Its overactivation leads to massive NAD+ consumption and ATP depletion. The ischemia/reperfusion injury (IR) induces PARP-1 overactivation and leads to cellular necrosis by massive ATP consumption. Our working hypothesis was that massive PARP-1 tubular expression in allograft human kidneys are a direct cause of acute tubular necrosis (ATN) and contribute to delay in early recovery of renal function (RRF) of the transplanted organ. Material and Methods:A total of 193 paraffin embedded renal allograft biopsies (95 pre-implant donor biopsies and 98 post-implant) with several ATN degrees, and 65 control renal biopsies from donors without ATN were studied. ATN degree was classified as: Absence (0); mild (1) [50%]. Nuclear expression of PARP-1 in tubular cells was evaluated by immunohistochemistry using polymer-conjugate MasVision kit and the monoclonal antibody anti-PARP-1 (clone PAR01). It was semiquantitatively determined, and scored from 0 to 3. Results: The nuclear PARP-1 preceded the morphological features of ATN. Immunostaining was located mainly in tubular cells nuclei, in cases of intense injury also was observed in glomeruli (capillary endothelial cells and epithelial cells of Bowmans capsule). Immunostaining was observed until advanced ATN condition. All 95 pre-transplant renal biopsies with ATN degree 1 (86%) or degree 2 (14%) showed tubular nuclei PARP-1 expression. The 98 post-transplant biopsies with ATN showed more intense expression of PARP-1 [degree 2 (45%), degree 3 (25%)]. Statistically significant relationship between ATN degree and PARP-1 expression was found (r=0.565, p=0.0001, Pearson test), with a mean expression of 2.74±0.45 in sever ATN cases versus 1.94±0.74 in mild ATN cases, and 0.29±0.45 in non-ATN cases (p=0.0001, one way ANOVA test). In conclusion, PARP-1 are linked to induction of ATN, and plays an important role in early graft renal function. This fact is indicated by the stati! stically significant relation with delay in total RRF (r=0.578, p=0.0001, Pearson test), creatinine serum levels at biopsy time (r=0.649) and at 3 months (r=0.363, p=0.0001, Pearson test), but not at 6 or 12 months post-transplant
Asunto(s)
Humanos , Poli(ADP-Ribosa) Polimerasas/metabolismo , Necrosis Tubular Aguda/enzimología , Poli(ADP-Ribosa) Polimerasas/farmacología , Necrosis Tubular Aguda/patología , Riñón/enzimología , Riñón/patología , Trasplante de Riñón , Inmunohistoquímica/métodosRESUMEN
Mitogen-activated protein kinases (MAPK) may be involved in the pathogenesis of acute renal failure. This study investigated the expression of p-p38 MAPK and nuclear factor kappa B (NF-kappaB) in the renal cortex of rats treated with gentamicin. Twenty rats were injected with gentamicin, 40 mg/kg, i.m., twice a day for 9 days, 20 with gentamicin + pyrrolidine dithiocarbamate (PDTC, an NF-kappaB inhibitor), 14 with 0.15 M NaCl, i.m., twice a day for 9 days, and 14 with 0.15 M NaCl , i.m., twice a day for 9 days and PDTC, 50 mg kg(-1) day(-1), i.p., twice a day for 15 days. The animals were killed 5 and 30 days after the last of the injections and the kidneys were removed for histological, immunohistochemical and Western blot analysis and for nitrate determination. The results of the immunohistochemical study were evaluated by counting the p-p38 MAPK-positive cells per area of renal cortex measuring 0.05 mm2. Creatinine was measured by the Jaffé method in blood samples collected 5 and 30 days after the end of the treatments. Gentamicin-treated rats presented a transitory increase in plasma creatinine levels. In addition, animals killed 5 days after the end of gentamicin treatment presented acute tubular necrosis and increased nitrate levels in the renal cortex. Increased expression of p-p38 MAPK and NF-kappaB was also observed in the kidneys from these animals. The animals killed 30 days after gentamicin treatment showed residual areas of interstitial fibrosis in the renal cortex, although the expression of p-p38 MAPK in their kidneys did not differ from control. Treatment with PDTC reduced the functional and structural changes induced by gentamicin as well as the expression of p-p38 MAPK and NF-kappaB. The increased expression of p-p38 MAPK and NF-kappaB observed in these rats suggests that these signaling molecules may be involved in the pathogenesis of tubulointerstitial nephritis induced by gentamicin.
Asunto(s)
Antibacterianos/efectos adversos , Gentamicinas/efectos adversos , Necrosis Tubular Aguda/enzimología , FN-kappa B/metabolismo , Nefritis Intersticial/enzimología , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismo , Animales , Western Blotting , Creatinina/sangre , Femenino , Fibrosis/enzimología , Fibrosis/patología , Inmunohistoquímica , Corteza Renal/química , Corteza Renal/efectos de los fármacos , Corteza Renal/patología , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/patología , Nefritis Intersticial/inducido químicamente , Nefritis Intersticial/patología , Nitratos/análisis , Pirrolidinas/farmacología , Ratas , Ratas Wistar , Tiocarbamatos/farmacologíaRESUMEN
Mitogen-activated protein kinases (MAPK) may be involved in the pathogenesis of acute renal failure. This study investigated the expression of p-p38 MAPK and nuclear factor kappa B (NF-kappaB) in the renal cortex of rats treated with gentamicin. Twenty rats were injected with gentamicin, 40 mg/kg, im, twice a day for 9 days, 20 with gentamicin + pyrrolidine dithiocarbamate (PDTC, an NF-kappaB inhibitor), 14 with 0.15 M NaCl, im, twice a day for 9 days, and 14 with 0.15 M NaCl , im, twice a day for 9 days and PDTC, 50 mg kg-1 day-1, ip, twice a day for 15 days. The animals were killed 5 and 30 days after the last of the injections and the kidneys were removed for histological, immunohistochemical and Western blot analysis and for nitrate determination. The results of the immunohistochemical study were evaluated by counting the p-p38 MAPK-positive cells per area of renal cortex measuring 0.05 mm². Creatinine was measured by the Jaffé method in blood samples collected 5 and 30 days after the end of the treatments. Gentamicin-treated rats presented a transitory increase in plasma creatinine levels. In addition, animals killed 5 days after the end of gentamicin treatment presented acute tubular necrosis and increased nitrate levels in the renal cortex. Increased expression of p-p38 MAPK and NF-kappaB was also observed in the kidneys from these animals. The animals killed 30 days after gentamicin treatment showed residual areas of interstitial fibrosis in the renal cortex, although the expression of p-p38 MAPK in their kidneys did not differ from control. Treatment with PDTC reduced the functional and structural changes induced by gentamicin as well as the expression of p-p38 MAPK and NF-kappaB. The increased expression of p-p38 MAPK and NF-kappaB observed in these rats suggests that these signaling molecules may be involved in the pathogenesis of tubulointerstitial nephritis induced by gentamicin.
Asunto(s)
Animales , Femenino , Ratas , Antibacterianos/efectos adversos , Gentamicinas/efectos adversos , Necrosis Tubular Aguda/enzimología , FN-kappa B/metabolismo , Nefritis Intersticial/enzimología , /metabolismo , Western Blotting , Creatinina/sangre , Fibrosis/enzimología , Fibrosis/patología , Inmunohistoquímica , Corteza Renal/química , Corteza Renal/efectos de los fármacos , Corteza Renal/patología , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/patología , Nefritis Intersticial/inducido químicamente , Nefritis Intersticial/patología , Nitratos/análisis , Pirrolidinas/farmacología , Ratas Wistar , Tiocarbamatos/farmacologíaRESUMEN
PURPOSE: In this research ACE activity (as a marker of epithelial injury) was studied in rats with gentamicin induced renal toxicity. METHODS: Male Sprague-Dawley rats were sacrificed 1, 3, 5, and 7 days after gentamicin injection, 100 mg/kg/day for 1, 3, 5, and 7 consecutive days. ACE activity was measured in serum, kidney and lung. These data were compared with normal saline-treated rats. Histological scoring of renal cortical pathology was performed on days 1, 3, 5, and 7. RESULTS: Treatment of rats with gentamicin resulted in renal damage evidenced by proteinuria, polyuria, and decreased creatinine clearance. The damage to the kidney proximal tubule was evident by (a) the histological analysis at light microscopy and (b) the augmentation in the urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG). Kidney ACE activity decreased while lung and serum ACE activity didn't change until day 7. Lung ACE activity increased significantly on day 7. Kidney and serum ACE activity increased too. Blood pressure increased significantly on day 7. This corresponded well with the lung ACE activity increment. CONCLUSION: These data suggest that kidney ACE activity decreased significantly just one day after gentamicin administration and prior to kidney NAG decrease.
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Lesión Renal Aguda/enzimología , Peptidil-Dipeptidasa A/metabolismo , Lesión Renal Aguda/fisiopatología , Análisis de Varianza , Animales , Biomarcadores/análisis , Biopsia con Aguja , Cromatografía Líquida de Alta Presión , Modelos Animales de Enfermedad , Activación Enzimática/fisiología , Gentamicinas/toxicidad , Inmunohistoquímica , Pruebas de Función Renal , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/fisiopatología , Masculino , Peptidil-Dipeptidasa A/análisis , Probabilidad , Ratas , Ratas Sprague-Dawley , Sensibilidad y EspecificidadRESUMEN
Our earlier studies with S-(1,2-dichlorovinyl)-L-cysteine (DCVC) showed that prior administration of a low priming dose of 15 mg/kg, i.p. to mice, given 72 hours before administration of a normally lethal dose of DCVC (75 mg/kg, i.p.) led to renal tubule necrosis, however sustained renal tubule regeneration was observed and these mice recovered from renal failure and survived. The objective of the present study was to investigate the role of extracellular signal-regulated kinase (ERK) pathway in this autoprotection model. Following the priming dose of DCVC, IL-6 protein and mRNA increased markedly as early as 1 hour after dosing, peaking at 3 hours with a 1.5-fold increase in plasma. Immunocytochemistry on kidney sections using specific antibodies against TGF-alpha, HB-EGF, EGFr, IGF-1Rbeta, Grb-2, and phospho-p44/42 MAP kinase (ERK1/2) revealed a significantly higher staining of these molecules 3 to 72 hours after dosing, indicating up regulation of the ERK pathway. Following a lethal dose of DCVC (75 mg/kg) the early increase in these signaling molecules was not sustained, being markedly reduced 24 and 36 hours after dosing, leading to inhibition of S-phase DNA synthesis, cell division and renal tubule repair. In contrast, prior treatment with a low dose of DCVC, followed by a high dose led to a sustained stimulation of the renal ERK pathway, renal tubule regeneration and recovery from acute renal failure. These results suggest that a sustained activation of the ERK1/2 pathway may be a key factor in enabling a continued renal tubule repair and hence protection from the progressive phase of DCVC-induced acute renal tubular necrosis in the mouse.
Asunto(s)
Cisteína/análogos & derivados , Necrosis Tubular Aguda/enzimología , Riñón/enzimología , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Recuperación de la Función/fisiología , Animales , Biomarcadores/análisis , Cisteína/toxicidad , Replicación del ADN/efectos de los fármacos , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente Indirecta , Técnicas para Inmunoenzimas , Interleucina-6/metabolismo , Riñón/efectos de los fármacos , Riñón/patología , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/patología , Masculino , Ratones , Proteína Quinasa 1 Activada por Mitógenos/genética , Proteína Quinasa 3 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/genética , ARN Mensajero/metabolismo , Recuperación de la Función/efectos de los fármacos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Fase S/efectos de los fármacosRESUMEN
BACKGROUND: Renal failure and toxic acute tubular necrosis (ATN) may be seen following exposure to a variety of therapeutic agents. Zoledronate (Zometa) is a new, highly potent bisphosphonate used in the treatment of hypercalcemia of malignancy. We report the first clinical-pathologic study of nephrotoxicity associated with this agent. METHODS: A cohort of six patients (four males and two females) with a mean age of 69.2 years received bisphosphonate therapy for multiple myeloma (five patients) or Paget's disease (one patient). In all patients, zoledronate was administered at a dose of 4 mg intravenously monthly, infused over at least 15 minutes, and the duration of therapy was mean 4.7 months (range, 3 to 9 months). RESULTS: All patients developed renal failure with a rise in serum creatinine from a mean baseline level of 1.4 mg/dL to 3.4 mg/dL. Renal biopsy revealed toxic ATN, characterized by tubular cell degeneration, loss of brush border, and apoptosis. Immunohistochemical staining revealed a marked increase in cell cycle-engaged cells (Ki-67 positive) and derangement in tubular Na+,K+-ATPase expression. Importantly, although all patients had been treated with pamidronate prior to zoledronate, no biopsy exhibited the characteristic pattern of collapsing focal segmental glomerulosclerosis observed in pamidronate nephrotoxicity. Following renal biopsy, treatment with zoledronate was discontinued and all six patients had a subsequent improvement in renal function (mean final serum creatinine, 2.3 mg/dL at 1 to 4 months of follow-up). CONCLUSION: The close temporal relationship between zoledronate administration and the onset of renal failure and the partial recovery of renal function following drug withdrawal strongly implicate this important and widely used agent in the development of toxic ATN.
Asunto(s)
Difosfonatos/efectos adversos , Imidazoles/efectos adversos , Necrosis Tubular Aguda/inducido químicamente , Anciano , Anciano de 80 o más Años , Apoptosis , Biopsia , Estudios de Cohortes , Femenino , Humanos , Riñón/patología , Riñón/fisiopatología , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/patología , Necrosis Tubular Aguda/fisiopatología , Masculino , Persona de Mediana Edad , Insuficiencia Renal/inducido químicamente , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Factores de Tiempo , Ácido ZoledrónicoRESUMEN
BACKGROUND: Early detection of acute tubular necrosis (ATN) could permit implementation of salvage therapies and improve patient outcomes in acute renal failure (ARF). The utility of single and combined measurements of urinary tubular enzymes in predicting ARF in critically ill patients has not been evaluated using the receiver-operating characteristic (ROC) plot method. METHODS: In this prospective pilot study, 26 consecutive critically ill adult patients admitted to the intensive-care unit were studied. Urine samples were collected twice daily for up to 7 days. ARF was defined as an increase in plasma creatinine of > or = 50% and > or = 0.15 mmol/l. ROC plot analysis was applied to the tubular marker data to derive optimum cut-offs for ARF. RESULTS: Four of the 26 study subjects (15.4%) developed ARF. Indexed to urinary creatinine concentration, gamma glutamyl transpeptidase (gamma GT), alkaline phosphatase (AP), N-acetyl-glucosaminidase (NAG), and alpha- and pi-glutathione S-transferase (alpha- and pi-GST) but not lactate dehydrogenase (LDH) were higher in the ARF group on admission (P<0.05). gamma GT, and alpha- and pi-GST remained elevated at 24 h. The onset of ARF based on changes in plasma creatinine varied from 12 h to 4 days (median 36 h). ROC plot analysis showed that gamma GT, pi-GST, alpha-GST, AP and NAG had excellent discriminating power for ARF (AUC 0.950, 0.929, 0.893, 0.863 and 0.845, respectively). The discriminating strength of creatinine clearance, while lower, was still significant (AUC 0.796). Positive and negative predictive values for ARF on admission were 67/100% for gamma GT, 67/90% for AP, 60/95% for alpha-GST, and 67/100% for pi-GST indices. Positive and negative predictive values for ARF for creatinine clearance < or = 23 ml/min were 50 and 91%, respectively. Creatinine clearances tended to be lower in ARF than in non-ARF patients on admission (P=0.06) and were significantly lower (P=0.008) after 12 h. Plasma urea and fractional sodium excretion were unhelpful. CONCLUSIONS: Tubular enzymuria on admission to the ICU is useful in predicting ARF. The cheapness and wide availability of automated assays for gamma GT and AP suggests that estimation of these enzymes in random urine samples may be particularly useful for identifying patients at high risk of ARF.
Asunto(s)
Acetilglucosaminidasa/orina , Lesión Renal Aguda/etiología , Lesión Renal Aguda/metabolismo , Fosfatasa Alcalina/orina , Glutatión Transferasa/orina , Unidades de Cuidados Intensivos , Necrosis Tubular Aguda/complicaciones , Necrosis Tubular Aguda/metabolismo , L-Lactato Deshidrogenasa/orina , gamma-Glutamiltransferasa/orina , Lesión Renal Aguda/prevención & control , Adolescente , Adulto , Anciano , Creatinina/sangre , Creatinina/orina , Femenino , Humanos , Necrosis Tubular Aguda/enzimología , Masculino , Persona de Mediana Edad , Proyectos Piloto , Valor Predictivo de las Pruebas , Estudios Prospectivos , Curva ROC , Sodio/sangre , Sodio/orina , Factores de TiempoRESUMEN
Ischemia reperfusion injury (IRI) is a major cause of delayed graft function. Recent studies have shown that selectins play an important role in IRI. Selectins bind to sialylated and fucosylated sLe(x) receptors, and two enzymes, fucosyltransferase IV (FucT-IV) and VII (FucT-VII), are important in the function of these receptors. We hypothesized that fucosyltransferase (FucT) enzymes were important pathophysiologic mediators of renal IRI. We therefore evaluated renal IRI in mice deficient in FucT-IV, FucT-VII, and both FucT-IV and FucT-VII and compared their renal function, tubular injury, selectin ligand expression, and neutrophil infiltration to those in wild-type control mice. Bilateral 30-min renal IRI was performed, and the results demonstrated that mice deficient in both FucT-IV/FucT-VII were significantly protected from renal IRI at 24 and 48 h compared with wild-type control mice. FucT-IV-deficient mice showed only modest protection from renal injury at 24 h. However, FucT-VII-deficient mice had similar injury as wild-type mice. Histological analysis of kidney tissue postischemia revealed that mice deficient in both FucT-IV and FucT-VII had significantly reduced tubular injury compared with wild-type mice. Selectin ligand expression increased postischemia in wild-type, but not FucT-IV/FucT-VII-deficient, mice. Neutrophil infiltration in postischemic kidneys of FucT-IV/FucT-VII-deficient mice was also attenuated. These data demonstrate that fucosyltransferases are important in the pathogenesis of renal IRI and are potential therapeutic targets.
Asunto(s)
Fucosiltransferasas/fisiología , Riñón/enzimología , Riñón/fisiopatología , Daño por Reperfusión/enzimología , Daño por Reperfusión/fisiopatología , Animales , Creatinina/sangre , Selectina E/análisis , Selectina E/biosíntesis , Selectina E/genética , Fucosiltransferasas/deficiencia , Fucosiltransferasas/genética , Humanos , Inmunoglobulina M/genética , Riñón/irrigación sanguínea , Riñón/patología , Pruebas de Función Renal , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/genética , Necrosis Tubular Aguda/prevención & control , Recuento de Leucocitos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Infiltración Neutrófila/genética , Neutrófilos/patología , Proteínas Recombinantes de Fusión/análisis , Daño por Reperfusión/genética , Daño por Reperfusión/patología , Coloración y EtiquetadoRESUMEN
We have shown previously that gamma-glutamyl transpeptidase (GGT) activity is essential for the nephrotoxicity of cisplatin. In this study we asked whether GGT activity was necessary for the antitumor activity of cisplatin. GGT was transfected into PC3 cells, a human prostate tumor cell line. Two independent GGT-positive cell lines were isolated and characterized. GGT cleaves extracellular glutathione providing the cells with access to additional cysteine. Expression of GGT had no effect on the growth rate of the cells in vitro where the culture medium contains high levels of cysteine. However, when the cells were injected into nude mice the GGT-positive tumors grew at more than twice the rate of the GGT-negative tumors. Weekly treatment with cisplatin was toxic to both GGT-positive and -negative tumors. The GGT-positive tumors were significantly more resistant to the toxicity of cisplatin than the GGT-negative tumors. Therefore, expression of GGT is required for the nephrotoxicity of cisplatin, but diminishes the tumor toxicity of the drug. These results indicate that the nephrotoxicity and the tumor toxicity of cisplatin are via two distinct pathways.
Asunto(s)
Antineoplásicos/farmacología , Cisplatino/farmacología , Cisteína/metabolismo , Resistencia a Antineoplásicos , Glutatión/metabolismo , gamma-Glutamiltransferasa/fisiología , Animales , Antineoplásicos/efectos adversos , Antineoplásicos/farmacocinética , Antineoplásicos/uso terapéutico , Cisplatino/efectos adversos , Cisplatino/farmacocinética , Cisplatino/uso terapéutico , Medios de Cultivo , Humanos , Necrosis Tubular Aguda/inducido químicamente , Necrosis Tubular Aguda/enzimología , Túbulos Renales Proximales/efectos de los fármacos , Túbulos Renales Proximales/enzimología , Masculino , Ratones , Ratones Desnudos , Neoplasias de la Próstata/patología , Proteínas Recombinantes de Fusión/fisiología , Transfección , Células Tumorales Cultivadas/efectos de los fármacos , gamma-Glutamiltransferasa/deficiencia , gamma-Glutamiltransferasa/genéticaRESUMEN
UNLABELLED: Glutathione S-Tranferases (GST) are the enzymes which are strictly specific for epithelial cells of the proximal and distal tubules in the kidney. These enzymes are detected in the urine when some tubular damage process is found. In healthy people urine GST is hardly detected. The goal of this study was to evaluate the release of two isoenzymes -- alpha and pi GST in the urine of kidney graft recipients during the first week after kidney transplantation, aiming to differentiate the cause of the delayed function (DF) of transplanted kidney. MATERIAL AND METHODS: 50 cadaveric kidneys were procured using standard technique with "in situ" cooling using UW solution. All kidneys were machine perfused. After preservation kidneys were transplanted to 50 ERSD patients. Standard triple drug immunosuppression was applied (steroids, CsA, Cell-Cept or Aza.). Graft function and the release of alpha and pi GST in the urine were measured 1, 3 and 7 days after transplantation. RESULTS: immediate function (IF) was found in 72% (36pts), DF in 28% (14pts). 5 of DF patients had ATN, 4 had acute rejection (REJ) and the remaining 5 had ATN and acute rejection (see table below). CONCLUSIONS: High alpha and pi GST concentrations were found in pts with DF graft function during the first 7 days after Tx. Elevated pi GST and low alpha GST in the urine indicates acute rejection. High alpha and pi GST in pts with DF should raise suspicion of graft rejection.
Asunto(s)
Glutatión Transferasa/orina , Trasplante de Riñón/fisiología , Ciclosporina/efectos adversos , Gutatión-S-Transferasa pi , Rechazo de Injerto/diagnóstico , Rechazo de Injerto/enzimología , Humanos , Inmunosupresores/efectos adversos , Isoenzimas/orina , Fallo Renal Crónico/enzimología , Fallo Renal Crónico/cirugía , Trasplante de Riñón/efectos adversos , Necrosis Tubular Aguda/diagnóstico , Necrosis Tubular Aguda/enzimología , Necrosis Tubular Aguda/etiología , Factores de TiempoRESUMEN
To examine the mechanisms involved in the progression of mercury chloride (HgCl2)-induced acute tubular necrosis (ATN), we investigated the histopathological changes and the expression of inducible nitric oxide synthase (iNOS) mRNA and protein in renal cortices of rats at 20 hours after exposure to HgCl2. The expression of iNOS mRNA was significantly augmented in renal cortices of rats with HgCl2-induced acute renal failure (ARF). Likewise, the induction of iNOS protein was observed in damaged proximal tubule epithelial cells of rats with HgCl2-induced ARF. Pretreatment of rats with iNOS inhibitor aminoguanidine, however, suppressed the development of proximal tubule epithelial cell injury and prevented an increase in blood urea nitrogen and serum creatinine as well as resulting in a marked fall in iNOS mRNA and protein in rats with HgCl2-induced ARF. These observations indicate that the induction of iNOS may play a role in the progression of HgCl2-induced ATN through the exacerbation of proximal tubule epithelial cell damage.
Asunto(s)
Lesión Renal Aguda/inducido químicamente , Necrosis Tubular Aguda/inducido químicamente , Cloruro de Mercurio/toxicidad , Óxido Nítrico Sintasa/metabolismo , Lesión Renal Aguda/enzimología , Animales , Femenino , Necrosis Tubular Aguda/enzimología , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Serum IgG antibodies against Tamm-Horsfall protein and urinary excretion of NAG and alpha-1-microglobulin were measured in 26 patients with a Kock reservoir for continent urinary diversion or orthotopic bladder reconstruction in order to detect any signs of tubular damage. None of these markers for tubular damage was correlated to the postoperative observation time ranging between 2 and 16 years. No correlation was found between these markers and signs of renal scarring or upper urinary tract dilatation as judged from urographies. A positive correlation was demonstrated between NAG excretion and antibodies against Tamm-Horsfall protein. The annual reduction in GFR was increased in patients with elevated alpha-1-microglobulin excretion but not in patients with elevated titres of antibodies against Tamm-Horsfall protein or increased NAG excretion. Patients with previous or present reflux nipple problems had elevated excretion of alpha-1-microglobulin. Regular determinations of alpha-1-microglobulin excretion appear to be of value in the follow-up of these patients.
Asunto(s)
Acetilglucosaminidasa/orina , alfa-Globulinas/orina , Inmunoglobulina G/sangre , Necrosis Tubular Aguda/diagnóstico , Mucoproteínas/inmunología , Complicaciones Posoperatorias/diagnóstico , Derivación Urinaria , Reservorios Urinarios Continentes , Adulto , Anciano , Cistectomía , Diagnóstico Diferencial , Femenino , Estudios de Seguimiento , Tasa de Filtración Glomerular/fisiología , Humanos , Pruebas de Función Renal , Necrosis Tubular Aguda/enzimología , Masculino , Persona de Mediana Edad , Complicaciones Posoperatorias/enzimología , Neoplasias de la Vejiga Urinaria/cirugía , Vejiga Urinaria Neurogénica/cirugía , Uromodulina , Reflujo Vesicoureteral/diagnóstico , Reflujo Vesicoureteral/enzimologíaRESUMEN
BACKGROUND: Carbon dioxide absorbents degrade sevoflurane, particularly at low gas flow rates, to fluoromethyl-2,2-difluoro-1-(trifluoromethyl)vinyl ether (compound A). Compound A causes renal proximal tubular injury in rats but has had no effect on blood urea nitrogen (BUN) or creatinine concentrations in patients. This investigation compared the effects of low-flow sevoflurane and isoflurane on renal tubular function in surgical patients using conventional (BUN and creatinine) and finer indices of renal injury, specifically those biomarkers sensitive for compound A toxicity in rats (glucosuria, proteinuria, and enzymuria [N-acetyl-beta-D-glucosaminidase (NAG) and alpha-glutathione-S-transferase (alpha GST)]). METHODS: Consenting patients with normal preoperative renal function at two institutions were randomized to receive sevoflurane (n = 36) or isoflurane (n = 37) in oxygen and air. Total gas flow was 1 l/min, opioid doses were minimized, and barium hydroxide lime was used to maximize anesthetic degradation. Inspiratory and expiratory compound A concentrations were quantified every 30-60 min. Blood and urine were obtained before and 24-72 h after anesthesia for laboratory evaluation. RESULTS: Sevoflurane and isoflurane groups were similar with respect to age, weight, sex, American Society of Anesthesiologists status, anesthetic duration (3.7 or 3.9 h), and anesthetic exposure (3.6 or 3 minimum alveolar concentration [MAC]-hour). Maximum inspired compound A concentration (mean +/- standard deviation) was 27 +/- 13 ppm (range, 10-67 ppm). Areas under the inspired and expired compound A concentration versus time curves (AUC) were 79 +/- 54-ppm-h (range, 10-223 ppm-h) and 53 +/- 40 ppm-h (range, 6-159 ppm-h), respectively. There was no significant difference between anesthetic groups in postoperative serum creatinine or BUN, or urinary excretion of protein, glucose, NAG, proximal tubular alpha GST, or distal tubular pi GST. There was no significant correlation between compound A exposure (AUC) and protein, glucose, NAG, alpha GST, or pi GST excretion. Postoperative alanine and aspartate aminotransferase concentrations were not different between the anesthetic groups, and there were no significant correlations between compound A exposure and alanine or aspartate aminotransferase concentrations. CONCLUSIONS: The renal tubular and hepatic effects of low-flow sevoflurane and isoflurane were similar as assessed using both conventional measures of hepatic and renal function and more sensitive biochemical markers of renal tubular cell necrosis. Moderate duration low-flow sevoflurane anesthesia, during which compound A formation occurs, appears to be as safe as low-flow isoflurane anesthesia.