RESUMEN
Regulatory T cells (Tregs ) have been recognized as central mediators for maintaining peripheral tolerance and limiting autoimmune diseases. The loss of Tregs or their function has been associated with exacerbation of autoimmune disease. However, the temporary loss of Tregs in the chronic spontaneous disease model has not been investigated. In this study, we evaluated the role of Tregs in a novel chronic spontaneous glomerulonephritis model of B cell lymphoma 2-interacting mediator (Bim) knock-out mice by transient depleting Tregs . Bim is a pro-apoptotic member of the B cell lymphoma 2 (Bcl-2) family. Bim knock-out (Bim-/- ) mice fail to delete autoreactive T cells in thymus, leading to chronic spontaneous autoimmune kidney disease. We found that Treg depletion in Bim-/- mice exacerbated the kidney injury with increased proteinuria, impaired kidney function, weight loss and greater histological injury compared with wild-type mice. There was a significant increase in interstitial infiltrate of inflammatory cells, antibody deposition and tubular damage. Furthermore, the serum levels of cytokines interleukin (IL)-2, IL-4, IL-6, IL-10, IL-17α, interferon (IFN)-γ and tumour necrosis factor (TNF)-α were increased significantly after Treg depletion in Bim-/- mice. This study demonstrates that transient depletion of Tregs leads to enhanced self-reactive T effector cell function followed by exacerbation of kidney disease in the chronic spontaneous kidney disease model of Bim-deficient mice.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Proteína 11 Similar a Bcl2/genética , Glomerulonefritis/inmunología , Depleción Linfocítica , Linfocitos T Reguladores/inmunología , Animales , Enfermedades Autoinmunes/patología , Proteína 11 Similar a Bcl2/deficiencia , Citocinas/sangre , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Glomerulonefritis/patología , Glomerulonefritis/fisiopatología , Interleucina-10/sangre , Interleucina-6/sangre , Riñón/inmunología , Riñón/patología , Riñón/fisiopatología , Ratones , Ratones Noqueados , ProteinuriaRESUMEN
AIM: While initial research suggests that M2 macrophages are athero-protective, more recently, proatherogenic functions, such as a greater uptake of lipid than M1 macrophages, have been demonstrated, raising the question of their actual association with plaque stability. The present study, therefore, assessed the association between macrophage subset and plaque stability. Furthermore, it examined whether the fibrocyte, that we have previously identified in the plaque, represents a subset of M2 macrophages. METHODS: Twenty human carotid atherosclerotic plaque specimens were examined for the presence of macrophages using immunohistochemistry for pan macrophages (CD68), M1 (CD64, CD86) and M2 (CD163, CD206) subsets. The slides were assessed by digital whole slide scanning/image analysis to quantify the expression of these markers in the plaque. Comparisons in marker distribution and quantity relative to plaque stability were made. Adoption of a fibrocyte phenotype was assessed by double immunofluorescence staining of the markers with procollagen I. RESULTS: M1 and M2 macrophages were present throughout the plaque including the core and cap. While the levels of CD68 (pan macrophage maker) and CD86 negatively correlated with cap thickness, the levels of the M2 marker, CD163, did not and moreover, did not differ between plaques when they were separated into stable and unstable groups. Notably, collagen production was evident in most but not all M2 macrophages. CONCLUSION: Our findings demonstrate that while macrophage levels in general negatively correlate with plaque cap thickness, levels of M2 macrophages do not. This may be in part due to their ability to produce collagen (ie adopt a fibrocyte phenotype) in the plaque.
Asunto(s)
Enfermedades de las Arterias Carótidas/patología , Macrófagos/patología , Placa Aterosclerótica/patología , Enfermedades de las Arterias Carótidas/cirugía , Endarterectomía Carotidea , Humanos , Placa Aterosclerótica/inmunología , Estudios RetrospectivosRESUMEN
BACKGROUND AND AIMS: Studies of diet in relation to chronic kidney disease (CKD) have focused on individual nutrients. The relationship between overall patterns of food intake and renal function has not been well explored. We aimed to investigate the associations between diet quality with the prevalence, incidence and progression of CKD in a cohort of older adults. METHODS AND RESULTS: 1952 participants aged ≥50 years at baseline were examined between 1992-1994 and 2002-2004. Dietary data were collected using a semi-quantitative food frequency questionnaire. A modified version of the Healthy Eating Index for Australians was developed to determine total diet scores (TDS). Baseline biochemistry including serum creatinine was measured. CKD was defined as MDRD estimated glomerular filtration rate (eGFR) <60 mL min⻹·1.73 m⻲. Participants in the highest quartile of mean TDS compared to those in the first quartile (reference), had a 41% reduced likelihood of having eGFR <60 mL min⻹·1.73 m⻲, [multivariable-adjusted odds ratio, OR, 0.59 (95% confidence intervals, CI, 0.41-0.85), P-trend = 0.005]. Each unit increase in TDS was associated with a 15% decrease in the odds of having prevalent CKD, multivariable-adjusted OR 0.85 (95% CI 0.74-0.97). A non-significant, inverse association between TDS and CKD incidence was observed (P-trend = 0.10). CONCLUSION: Older adults with better diet quality had a reduced likelihood of having eGFR <60 mL min⻹·1.73 m⻲. Adherence to dietary guidelines were not prospectively associated with CKD incidence or progression. Further studies with adequate power are warranted to assess the longitudinal association between diet quality and CKD.
Asunto(s)
Envejecimiento , Dieta , Riñón/fisiopatología , Insuficiencia Renal Crónica/prevención & control , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Estudios Transversales , Dieta/efectos adversos , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Tasa de Filtración Glomerular , Promoción de la Salud , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Nueva Gales del Sur/epidemiología , Política Nutricional , Cooperación del Paciente , Prevalencia , Insuficiencia Renal Crónica/epidemiología , Insuficiencia Renal Crónica/etiología , Insuficiencia Renal Crónica/fisiopatología , Factores de Riesgo , Salud UrbanaRESUMEN
Based on new understanding of the diverse biological functions of macrophages and dendritic cells (DC), the focus of studies on these cells has been expanded from their pathogenic role in renal diseases to include their potential to regulate inflammation and restore renal architecture and function. By exploiting their regulatory function, macrophages or DC have been used to treat experimental renal disease following their adoptive transfer. This review summarizes current progress in the therapeutic use of macrophages and DC in renal diseases. Key issues for ongoing research are discussed.
Asunto(s)
Células Dendríticas/fisiología , Enfermedades Renales/terapia , Macrófagos/fisiología , Traslado Adoptivo , Animales , Terapia Genética , Glomerulonefritis/terapia , Enfermedades Renales/inmunología , Enfermedades Renales/prevención & controlRESUMEN
Lymphocytes and macrophages act as effector immune cells in the initiation and progression of renal injury. Recent data have shown that subpopulations of these immune cells (regulatory T lymphocytes and alternately-activated or regulatory macrophages) are potent modulators of tissue injury and repair in renal disease. Recent animal studies examining the therapeutic effect of these cells raise the exciting possibility that strategies targeting these cell types may be effective in treating and preventing kidney disease in humans. This review will describe their biological role in experimental kidney disease and therapeutic potential in clinical nephrology.
Asunto(s)
Enfermedades Renales/fisiopatología , Macrófagos/fisiología , Linfocitos T Reguladores/fisiología , Animales , Células Dendríticas/patología , Células Dendríticas/fisiología , Humanos , Enfermedades Renales/patología , Macrófagos/patología , Mastocitos/patología , Mastocitos/fisiología , Linfocitos T Reguladores/patologíaRESUMEN
Macrophage infiltration of the kidney is a prominent feature associated with the severity of renal injury and progressive renal failure. To determine the influence of macrophages in renal disease models in the absence of endogenous T and B cells, we performed adoptive transfer of macrophages into severe combined immunodeficient (SCID) mice. In this study, macrophages were isolated from the spleens of BALB/c mice and stimulated with lipopolysaccharide to induce classically activated M1 macrophages or with interleukin-4 (IL-4) and IL-13 to induce alternatively activated M2 macrophages. These macrophages were then infused into SCID mice with adriamycin nephropathy; an in vivo model of chronic inflammatory renal disease analogous to human focal segmental glomerulosclerosis. Mice infused with M1 macrophages had a more severe histological and functional injury, whereas M2 macrophage-induced transfused mice had reduced histological and functional injury. Both M1 and M2 macrophages localized preferentially to the area of injury and maintained their phenotypes even after 4 weeks. The protective effect of M2 macrophages was associated with reduced accumulation and possibly downregulated chemokine and inflammatory cytokine expression of the host infiltrating macrophages. Our findings demonstrate that macrophages not only act as effectors of immune injury but can be induced to provide protection against immune injury.
Asunto(s)
Glomeruloesclerosis Focal y Segmentaria/inmunología , Activación de Macrófagos , Macrófagos/fisiología , Traslado Adoptivo/métodos , Animales , Movimiento Celular/fisiología , Células Cultivadas , Quimiocina CCL17 , Quimiocinas CC/metabolismo , Modelos Animales de Enfermedad , Regulación hacia Abajo , Doxorrubicina , Glomeruloesclerosis Focal y Segmentaria/inducido químicamente , Glomeruloesclerosis Focal y Segmentaria/prevención & control , Interleucina-10/metabolismo , Interleucina-13/farmacología , Interleucina-4/farmacología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/trasplante , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones SCID , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fenotipo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Programmed death 1 (PD-1) is a novel member of the CD28/cytotoxic T-lymphocyte-associated protein-4 superfamily, which plays an important role in the regulation of activated T cells. However, it is not clear how PD-1 is expressed in normal and diseased kidney, nor if it has a role in progression of chronic renal disease. PD-1 expression and the effect of monoclonal anti-PD-1 antibody (Ab) were examined in murine adriamycin nephropathy (AN). BALB/c mice were divided into three groups: (a) normal mice, (b) adriamycin (ADR) with control immunoglobulin (Ig)G (ADR-IgG), and (c) ADR with anti-PD-1 Ab (ADR-Ab). AN was induced by a single intravenous injection of ADR. Anti-PD-1 Ab was given by intraperitoneal injection on alternate days from day 0 to day 10, or to day 18. Animals were killed at week 4. Renal function, histological change, and cytokine expression were examined. PD-1 mRNA was detected in kidney tissue of mice with AN in a dose- and time-dependent manner. PD-1 was mainly expressed on injured tubule cells and some interstitial cells, which co-stained with alpha-smooth muscle actin in AN, but not in normal kidney. Anti-PD-1 treatment up to day 18, but not to day 10, worsened glomerular and tubulointerstitial injury. The ratio of urinary protein/creatinine was significantly higher in ADR-Ab mice than ADR-IgG mice. The number of macrophages was significantly increased in ADR-Ab mice compared with ADR-IgG mice. Blockade of PD-1 worsened progressive renal histopathological and functional injury in murine AN. This suggests a possible protective role for PD-1 in chronic renal disease, and its potential as a treatment to slow disease progression.
Asunto(s)
Antígeno B7-1/inmunología , Doxorrubicina , Glomeruloesclerosis Focal y Segmentaria/patología , Fallo Renal Crónico/patología , Glicoproteínas de Membrana/inmunología , Péptidos/inmunología , Actinas/inmunología , Animales , Anticuerpos Monoclonales/inmunología , Antígeno B7-1/administración & dosificación , Antígeno B7-1/genética , Antígeno B7-1/fisiología , Antígeno B7-H1 , Creatinina/orina , Cricetinae , Interpretación Estadística de Datos , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Relación Dosis-Respuesta a Droga , Doxorrubicina/administración & dosificación , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Glomeruloesclerosis Focal y Segmentaria/inducido químicamente , Glomeruloesclerosis Focal y Segmentaria/genética , Glomeruloesclerosis Focal y Segmentaria/inmunología , Glomeruloesclerosis Focal y Segmentaria/metabolismo , Glomeruloesclerosis Focal y Segmentaria/terapia , Inmunoglobulina G/inmunología , Inmunoglobulinas/inmunología , Inmunohistoquímica , Inyecciones Intraperitoneales , Inyecciones Intravenosas , Fallo Renal Crónico/inducido químicamente , Fallo Renal Crónico/genética , Fallo Renal Crónico/inmunología , Fallo Renal Crónico/metabolismo , Fallo Renal Crónico/terapia , Glomérulos Renales/patología , Túbulos Renales/patología , Macrófagos , Glicoproteínas de Membrana/administración & dosificación , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiología , Ratones , Ratones Endogámicos C57BL , Péptidos/administración & dosificación , Péptidos/genética , Péptidos/fisiología , Proteinuria/diagnóstico , ARN Mensajero/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Coloración y Etiquetado/métodos , Factores de TiempoRESUMEN
In adriamycin nephropathy (AN), a model of chronic proteinuric renal injury, the absence of functional B and T cells with residual natural killer (NK) cells, and macrophages in severe combined immunodeficient (SCID) mice results in more severe disease than in immunocompetent mice. We have recently shown expression of the stimulatory NK cell molecule NKG2D and its ligand RAE-1 in the adriamycin (ADR) kidney. Therefore, we sought to determine the role of NK cells in AN. We used anti-asialo GM1 NK cell depletion in immunocompetent BALB/c mice with AN, and also compared AN in immunodeficient SCID mice and immunodeficient nonobese diabetic (NOD)-SCID mice (that have impaired NK cell function). The number of NK cells was increased in AN in BALB/c mice compared with normal controls. NK cell depletion or reduction of NK function in NOD-SCID mice did not affect the severity of disease. In both wild type and immunodeficient models, ADR upregulated RAE-1 in the kidney. High levels of Class I major histocompatibility complex molecules were found in both models of AN. In conclusion, NK cells do not play a significant role in AN.
Asunto(s)
Doxorrubicina/toxicidad , Riñón/patología , Células Asesinas Naturales/inmunología , Animales , Citometría de Flujo , Antígenos de Histocompatibilidad Clase I/inmunología , Riñón/efectos de los fármacos , Riñón/inmunología , Pruebas de Función Renal , Células Asesinas Naturales/efectos de los fármacos , Depleción Linfocítica , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Ratones SCID , Subfamilia K de Receptores Similares a Lectina de Células NK , Reacción en Cadena de la Polimerasa , Proteinuria/etiología , Receptores Inmunológicos/genética , Receptores de Células Asesinas NaturalesRESUMEN
Over the next decade, the number of patients with end-stage renal disease (ESRD) treated by dialysis may double, and even developed nations will have difficulty in coping with this alarming increase. This review will outline the proven and unproven strategies that have the potential to retard the progression of chronic kidney disease (CKD). Recently, a number of randomised clinical trials have demonstrated the efficacy of several strategies to slow the progression of CKD. Proven strategies include adequate blood pressure control (with angiotensin blockade), and for diabetic nephropathy good glycaemic control. Other potentially beneficial strategies include smoking cessation, lipid control and aldosterone blockade. The early institution of these strategies has the potential to regress established CKD as well as improve the long-term cardiovascular outcomes of these patients. Proof of the efficacy in humans of promising experimental approaches, such as the administration of growth factors (e.g., recombinant bone morphogenetic protein-7), anti-fibrotic agents (e.g., pirfenidone) and novel anti-proteinuric drugs (e.g., pentosan polysulphate), is awaited. Finally, the primary prevention of CKD, at least in part, by the eradication of type 2 diabetes and obesity (through improvement of lifestyle factors), and adequate treatment of hypertension, have the potential to eliminate up to half of the most common causes of CKD (or ESRD) in developed countries.
Asunto(s)
Fallo Renal Crónico/prevención & control , Bloqueadores del Receptor Tipo 1 de Angiotensina II/uso terapéutico , Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Animales , Presión Sanguínea/efectos de los fármacos , Presión Sanguínea/fisiología , Dieta con Restricción de Proteínas , Progresión de la Enfermedad , Quimioterapia Combinada , Humanos , Hipertensión/complicaciones , Hipertensión/fisiopatología , Hipertensión/terapia , Fallo Renal Crónico/etiologíaRESUMEN
OBJECTIVE: To investigate the predictive value of quantitative measurements of blastocyst morphology on subsequent implantation rates after transfer. DESIGN: Prospective observational study. SETTING: Private assisted reproductive technology center. PATIENT(S): One hundred seventy-four IVF patients receiving transfers of expanded blastocyst-stage embryos on day 5 (n = 112) or day 6 (n = 62) after oocyte retrieval. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Blastocyst diameter, number of trophectoderm cells, inner cell mass (ICM) size, ICM shape, and implantation and pregnancy rates. RESULT(S): Blastocyst diameter and trophectoderm cell numbers were unrelated to implantation rates. Day 5 expanded blastocysts with ICMs of >4,500 microm(2) implanted at a higher rate than did those with smaller ICMs (55% vs. 31%). Day 5 expanded blastocysts with slightly oval ICMs implanted at a higher rate (58%) compared with those with either rounder ICMs (7%) or more elongated ICMs (33%). Implantation rates were highest (71%) for embryos with both optimal ICM size and shape. Pregnancy rates were higher for day 5 transfers of optimally shaped ICMs compared with day 5 transfers of optimally sized ICMs. CONCLUSION(S): Quantitative measurements of the inner cell mass are highly indicative of blastocyst implantation potential. Blastocysts with relatively large and/or slightly oval ICMs are more likely to implant than other blastocysts.
Asunto(s)
Blastocisto/citología , Implantación del Embrión/fisiología , Transferencia de Embrión , Fertilización In Vitro/métodos , Adulto , Blastocisto/fisiología , Tamaño de la Célula/fisiología , Femenino , Humanos , Masculino , Valor Predictivo de las Pruebas , Embarazo , Estudios Prospectivos , Estadísticas no ParamétricasRESUMEN
OBJECTIVE: To compare the outcome of second and third cycles of in vitro fertilization with blastocyst transfer to the outcome of first attempts at IVF with blastocyst transfer. DESIGN: Retrospective study. SETTING: Private ART center. PATIENT(S): Three hundred and four patients undergoing treatment with in vitro fertilization with blastocyst transfer, 87 of which underwent at least one cycle of re-treatment after failing to achieve pregnancy in their first cycle. INTERVENTION(S): Bipronucleate oocytes were grown for up to 144 hours and subsequently transferred when at least one embryo attained the expanded blastocyst stage. MAIN OUTCOME MEASURE(S): Pregnancy and implantation rates. RESULT(S): Pregnancy rates per retrieval were significantly higher for patients undergoing their first cycle of in vitro fertilization with blastocyst transfer (36%) compared to those undergoing their second (19%) or their third (9%) cycles of treatment. Implantation rates per embryo were also higher for first cycles of in vitro fertilization with blastocyst transfer (30%) compared to second (18%) or third cycles (8%). CONCLUSION(S): Pregnancy and implantation rates decline dramatically in repeated cycles of in vitro fertilization with blastocyst transfer following one or more unsuccessful cycles of in vitro fertilization with blastocyst transfer.
Asunto(s)
Implantación del Embrión , Transferencia de Embrión , Fertilización In Vitro , Embarazo , Adulto , Blastocisto , Femenino , Humanos , Resultado del Embarazo , Análisis de Regresión , Estudios Retrospectivos , Insuficiencia del TratamientoRESUMEN
Adriamycin nephropathy (AN) is a widely used nonimmune-mediated rat model of proteinuric chronic glomerular disease and is usually induced by a single intravenous injection of doxorubicin hydrochloride (DX) into the tail vein. However, this route can be associated with local skin necrosis and variability in disease induction and poses an occupational hazard to the investigator. Here we describe a simple technique of administering DX (1.7 mg) to ketamine:xylazine-sedated adult male Wistar rats (mean +/- 1 standard deviation, 238 +/- 8 g; n = 28) by using a single substernal intracardiac injection. The procedure was associated with minimal morbidity and mortality (1 death related to anesthesia). By day 21, severe nephrotic syndrome with effacement of glomerular epithelial cell foot processes and diffuse cortical tubulointerstitial injury was induced in all animals. Therefore, intracardiac injection of DX is a safe and consistent method of inducing AN in the rat and provides an alternative to the tail-vein route.
Asunto(s)
Doxorrubicina/administración & dosificación , Corazón/efectos de los fármacos , Glomérulos Renales/efectos de los fármacos , Síndrome Nefrótico/inducido químicamente , Proteinuria/inducido químicamente , Animales , Doxorrubicina/toxicidad , Riñón/patología , Glomérulos Renales/patología , Masculino , Microscopía Electrónica , Síndrome Nefrótico/patología , Ratas , Ratas Wistar , Cola (estructura animal)/irrigación sanguíneaRESUMEN
OBJECTIVE: To compare implantation and pregnancy rates according to the day of embryo transfer (day 5 or 6 after oocyte retrieval) when transfer was postponed until expanded blastocysts developed. DESIGN: Retrospective clinical study. SETTING: Private ART center. PATIENT(S): One-hundred and eighty-three women undergoing blastocyst-stage embryo transfer following in vitro fertilization. INTERVENTION(S): Bipronucleate oocytes were grown for up to 144 hours and subsequently transferred only when at least one embryo attained the expanded blastocyst stage. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): Blastocysts transferred on day 5 implanted at nearly twice the rate of blastocysts transferred on day 6 (36.3% vs. 19.0%). Pregnancy rates were also almost twice as high among the day 5 transfer patients (59.3% vs. 32.3%). In addition, more blastocysts developed (3.6 vs. 2.4), and more were transferred (2.7 vs. 2.3) to the day 5 transfer patients, although the proportion of expanded blastocysts among the blastocysts that were transferred was the same for the two groups (91.7% vs. 93.6%). CONCLUSION(S): Embryos that develop to the expanded blastocyst stage and are transferred on day 5 after retrieval are approximately twice as likely to implant compared to those for which expansion and transfer are delayed until day 6.
Asunto(s)
Transferencia de Embrión , Adulto , Blastocisto , Implantación del Embrión , Femenino , Fertilización In Vitro , Humanos , Embarazo , Índice de Embarazo , Estudios Retrospectivos , Factores de TiempoRESUMEN
Nitric oxide (NO) levels are increased after exposure of cultured proximal tubule cells (PTC) to non-haem iron, potentially contributing to PTC injury in disease states associated with increased iron exposure, including proteinuric renal disease. The mechanisms underlying this observed increase were investigated. After 3 h exposure to 400 microM nitrilotriacetate (NTA)-Fe, inducible nitric oxide synthase (iNOS) mRNA expression was significantly increased, with a corresponding increase in iNOS protein after 12 h. The nuclear binding activity of NFkappaB with 400 microM NTA-Fe was increased, and pyrrolidine dithiocarbamate (PDTC), an antioxidant inhibitor of NFkappaB, prevented both activation of NFkappaB and NO production in response to NTA-Fe. Inhibition of protein tyrosine kinase reduced iNOS mRNA, iNOS protein levels and NO production in response to NTA-Fe. The effect of tyrosine kinase inhibition on NFkappaB activation was variable, with herbimycin but not genistein having an inhibitory effect. Activation of either protein kinase A or C increased iNOS mRNA and protein levels, and NO production in response to NTA-Fe, whereas only the protein kinase C activator phorbol dibutyrate (PDBu) had a stimulatory effect on NFkappaB activation. The protein kinase A activator forskolin did not alter iron-induced activation of NFkappaB. These data suggest that the observed increase in NO production by PTC in response to iron is due to increased transcription of iNOS. The transcriptional regulation of this response is complex and involves NFkappaB, protein tyrosine kinase and the protein kinases A and C.
Asunto(s)
Colforsina/farmacología , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Compuestos Férricos/farmacología , Genisteína/farmacología , Hierro/metabolismo , Túbulos Renales Proximales/efectos de los fármacos , FN-kappa B/metabolismo , Óxido Nítrico Sintasa/metabolismo , Óxido Nítrico/biosíntesis , Ácido Nitrilotriacético/análogos & derivados , Ácido Nitrilotriacético/farmacología , Forbol 12,13-Dibutirato/farmacología , Proteína Quinasa C/metabolismo , Proteínas Tirosina Quinasas/antagonistas & inhibidores , Proteínas Tirosina Quinasas/metabolismo , Pirrolidinas/farmacología , Quinonas/farmacología , ARN Mensajero/metabolismo , Tiocarbamatos/farmacología , Animales , Benzoquinonas , Células Cultivadas , Proteína Quinasa Tipo II Dependiente de AMP Cíclico , Regulación Enzimológica de la Expresión Génica , Quelantes del Hierro/metabolismo , Túbulos Renales Proximales/metabolismo , Lactamas Macrocíclicas , Masculino , Óxido Nítrico/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II , ARN Mensajero/genética , Ratas , Ratas Wistar , Rifabutina/análogos & derivadosRESUMEN
Proximal tubule cells (PTC) in chronic renal disease produce chemokines which cause renal interstitial inflammation, and also transport more Na+ than normal. To investigate whether these two events might be related, monocyte chemoattractant protein-1 (MCP-1) production was examined in rat PTC in primary culture. Amiloride reduced, while ouabain increased levels of MCP-1 mRNA and protein. Amiloride reduced MCP-1 in cells stimulated with ouabain, lipopolysaccharide (LPS) or albumin. Intracellular Na+ rose with ouabain, but not LPS or albumin. Effects of amiloride, ouabain, LPS and albumin were abrogated by sodium-free but not chloride-free culture medium, and were not explained by changes in intracellular pH. Intracellular Ca2+ rose with ouabain, LPS or albumin and sodium-free medium. BAPTA-AM reduced intracellular Ca2+ and MCP-1 mRNA levels in unstimulated cells, and cells stimulated with ouabain, LPS or albumin. Thus, amiloride and ouabain may alter tubular cell MCP-1 by changing intracellular Na+, with secondary changes in intracellular Ca2+, whereas stimulation by LPS and albumin may involve Ca2+ directly.
Asunto(s)
Amilorida/farmacología , Calcio/metabolismo , Quimiocina CCL2/antagonistas & inhibidores , Ácido Egtácico/farmacología , Túbulos Renales Proximales/efectos de los fármacos , Lipopolisacáridos/farmacología , Ouabaína/farmacología , ARN Mensajero/metabolismo , Sodio/metabolismo , Equilibrio Ácido-Base/fisiología , Amilorida/análogos & derivados , Animales , Supervivencia Celular/fisiología , Células Cultivadas , Quimiocina CCL2/biosíntesis , Quimiocina CCL2/fisiología , Relación Dosis-Respuesta a Droga , Ácido Egtácico/análogos & derivados , Ensayo de Inmunoadsorción Enzimática , Colorantes Fluorescentes , Concentración de Iones de Hidrógeno , Túbulos Renales Proximales/metabolismo , Masculino , Ratas , Ratas Wistar , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ATPasa Intercambiadora de Sodio-Potasio/metabolismo , Espectrometría de FluorescenciaRESUMEN
Tubulointerstitial damage, in progressive chronic renal disease of all types, arises because of a complex interplay between factors in the tubular lumen, tubular epithelial cells, peritubular capillaries, resident and infiltrating interstitial cells and extracellular matrix. Particularly in proteinuric renal disease, tubular epithelial cells play a central role in orchestrating these events. In response to mediators arising systemically, in the tubular lumen or from other renal cells, tubular epithelial cells undergo a complex series of structural and functional changes and produce a bewildering number of soluble and fixed mediators, which in turn lead to interstitial inflammation and fibrosis. Knowledge of these interactions has increased exponentially over the past decade, and has defined a number of new targets for treatment. Both expansion and consolidation of this knowledge is needed to determine which of these targets holds the most promise for future treatment.
Asunto(s)
Nefritis Intersticial , Humanos , Nefritis Intersticial/metabolismo , Nefritis Intersticial/patología , Nefritis Intersticial/fisiopatologíaRESUMEN
BACKGROUND: Many studies have shown that interstitial inflammation in human and experimental renal disease is characterized by T-cell infiltration, but published data on the involvement of inflammatory cell subsets in progressive tubulointerstitial lesions are often conflicting. A previous study suggested a role for cytotoxic T lymphocytes in the damaging effect of CD4(+) T-cell depletion in murine adriamycin (ADR) nephropathy, a model of focal segmental glomerulosclerosis (FSGS), and tubulointerstitial inflammation. The aim of this study was to investigate the role of CD8(+) cells in this model. METHODS: Male BALB/c mice were treated with five intraperitoneal injections of anti-CD8 monoclonal antibody (mAb), beginning from five days after ADR treatment, when overt proteinuria was established. Seven mice in each of groups A (ADR + mAb), B (ADR only), and C (saline treated, age matched) were sacrificed at week 6. Changes in renal function and histopathological features were assessed. Tubulointerstitial inflammation and glomerular inflammation were examined immunohistochemically. RESULTS: mAb treatment reduced CD8(+) cell levels to <2% of normal in spleen. Proteinuria in group A was no different from that in group B at week 6, but was markedly higher than in group C. Creatinine clearance was significantly ameliorated by anti-CD8 treatment (71.8 +/- 4.9 microL/min vs. 29.2 +/- 2.8 in group B and 81.9 +/- 3.7 in group C). Morphometric analysis showed less FSGS in group A compared with group B (6.5 +/- 1.9 vs. 13.0 +/- 2.8, P < 0.001), as well as less tubular atrophy (indicated by increased ratio of tubule cell height to tubular diameter, 0.25 +/- 0.24 in group A vs. 0.04 +/- 0.02 in group B, P < 0.05). CD8 depletion also reduced interstitial expansion (6.3 +/- 2.2% vs. 16.4 +/- 3.1 in group B, P < 0.001) and fibrosis (P < 0.01). Macrophage infiltration in tubulointerstitium was less in group A than in group B (P = 0.052). The number of interstitial CD4(+) cells appeared to increase after anti-CD8 treatment, but was not statistically different between groups A and B. CONCLUSION: Anti-CD8 treatment protects against renal functional and structural injury in this murine model of chronic proteinuric renal disease.
Asunto(s)
Linfocitos T CD8-positivos/fisiología , Doxorrubicina , Enfermedades Renales/inducido químicamente , Enfermedades Renales/fisiopatología , Animales , Anticuerpos Monoclonales/farmacología , Atrofia , Linfocitos T CD8-positivos/inmunología , Creatinina/metabolismo , Progresión de la Enfermedad , Fibrosis , Glomeruloesclerosis Focal y Segmentaria/inducido químicamente , Glomeruloesclerosis Focal y Segmentaria/patología , Inmunohistoquímica , Enfermedades Renales/patología , Enfermedades Renales/orina , Túbulos Renales/patología , Ratones , Ratones Endogámicos BALB C , Proteinuria/etiologíaRESUMEN
BACKGROUND: CD4(+) T cells play an important role in various types of immunologic renal disease, including lupus nephritis, IgA nephropathy, and crescentic glomerulonephritis. CD4(+) T cells are also major infiltrating lymphocytes in chronic tubulointerstitial inflammation associated with nonimmunological renal diseases. We suspected that CD4(+) T cells might contribute to disease progression and loss of renal function in chronic proteinuric renal disease (CPRD). To investigate this possibility, the effect of monoclonal antibody against CD4(+) lymphocytes (anti-CD4) was studied in a murine model (adriamycin nephropathy) of CPRD. METHODS: Adriamycin nephropathy was produced in male BALB/c mice by a single intravenous injection of adriamycin (11 mg/kg). Anti-CD4 was given by intraperitoneal injection following the development of proteinuria at days 5, 6, 7, 21, and 37 after adriamycin. After six weeks, renal function and histology were studied by histomorphometry, immunohistochemistry, and flow cytometry. RESULTS: Flow cytometric analysis showed a marked decrease in the number of CD4(+) T cells in blood and spleen of the antibody-treated animals (N = 7, P < 0.01). Adriamycin plus CD4(+) depletion mice had significantly greater mesangial expansion, glomerular sclerosis, and interstitial expansion than the mice on adriamycin alone. Interstitial infiltration with macrophages and CD8(+) cells was significantly increased in adriamycin plus CD4(+) depletion mice. Creatinine clearance (17.5 +/- 0.54 vs. 29.2 +/- 0.89 microL/min, P < 0.001) was significantly worse in the adriamycin plus CD4(+) depletion mice than in adriamycin alone mice and correlated with histologic change in glomeruli and interstitium. CONCLUSIONS: Depletion of CD4(+) T cells promotes glomerular and interstitial injury in mice with established adriamycin nephropathy. These findings suggest that CD4(+) T cells have a protective role against the progression of adriamycin nephropathy.
Asunto(s)
Linfocitos T CD4-Positivos/fisiología , Doxorrubicina , Enfermedades Renales/inducido químicamente , Enfermedades Renales/patología , Glomérulos Renales/patología , Riñón/patología , Animales , Anticuerpos Monoclonales/farmacología , Recuento de Linfocito CD4 , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Interferón gamma/genética , Interleucina-4/genética , Riñón/metabolismo , Enfermedades Renales/metabolismo , Glomérulos Renales/metabolismo , Macrófagos/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Monocitos/patología , ARN Mensajero/metabolismoRESUMEN
The persistence of NF-kappaB independent inflammatory signals in the cortical tubulointerstitium may explain the incomplete suppression of interstitial monocyte accumulation by the antioxidant NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), in nephrotic rats with established Adriamycin nephropathy (AN). Because PDTC is known to have anti-proteinuric effects, in this study we investigated whether earlier commencement, during the pre-nephrotic phase of AN, would be more effective in reducing interstitial monocyte accumulation. Male Wistar rats with AN received either vehicle or PDTC (50 mg/kg bd i.p.i.) from d7 until d30 (n = 8 per group). On d30, PDTC reduced renal cortical lipid peroxidation (43%), wet kidney weight and tubulointerstitial injury in AN, but did not decrease proteinuria. Accordingly, inhibition of interstitial ED-1 accumulation remained incomplete (52%). Interestingly, the early administration of PDTC in AN, induced polyuria and renal cortical NF-kappaB DNA-binding activity was reduced by only 35%. These results suggest that: (i) the combination of an anti-proteinuric agent with PDTC may be required to completely suppress interstitial monocyte cell accumulation in AN and, (ii) the timing and duration of PDTC therapy are an important determinant of its efficacy to reduce NF-kappaB activation, in vivo.