RESUMO
AIMS: The study aimed to investigate the value of autoantibodies in predicting the risk of ketoacidosis or microalbuminuria in children with type 1 diabetes mellitus. METHODS: Clinical data and laboratory indicators of 80 patients with type 1 diabetes admitted to the Department of Endocrinology in Tianjin Children's Hospital, from June 2017 to March 2019, were retrospectively analyzed. The patients were divided into two groups: diabetes without ketoacidosis group (n = 20) and diabetes with ketoacidosis group (n = 60). The differences in general data, laboratory test indexes, and autoantibodies between the two groups were analyzed. Finally, ROC curves and multivariate logistic regression analysis were used to explore the value of autoantibodies in patients with ketoacidosis or microalbuminuria. RESULTS: A total of 80 children with type 1 diabetes were assessed, including 35 boys and 45 girls, ranging in age from 10 months to 15 years. The concentration of GADA, IA2A, and ZnT8A was not statistically different between the two groups, but the positive rate of ZnT8A was statistically significant (p = 0.038) and had a diagnostic value for the occurrence of ketoacidosis (p = 0.025). ZnT8A-positive patients had a higher titer of IA2A and a more frequent prevalence of GADA and IA2A than ZnT8A-negative patients (p < 0.01). In multivariate logistic regression analyses, the presence of positive ZnT8A was associated with a higher risk of microalbuminuria independent of age, sex, and BMI (OR = 4.184 [95% CI 1.034~16.934], p = 0.045). CONCLUSIONS: The positive ZnT8A had diagnostic value for ketoacidosis in children with type 1 diabetes and had the highest specificity among the three kinds of autoantibodies. Moreover, ZnT8A positivity was related to a higher titer of IA2A and more frequent occurrence of multiple diabetes-related autoantibodies. Besides, the presence of positive ZnT8A was an independent risk factor of microalbuminuria in children with type 1 diabetes. Therefore, we can infer that positive ZnT8A may be related to ketoacidosis and microalbuminuria, accelerating the progression of T1DM.
Assuntos
Albuminúria , Autoanticorpos/sangue , Diabetes Mellitus Tipo 1 , Cetose , Adolescente , Albuminúria/complicações , Albuminúria/epidemiologia , Albuminúria/imunologia , Criança , Pré-Escolar , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 1/epidemiologia , Diabetes Mellitus Tipo 1/imunologia , Feminino , Humanos , Lactente , Cetose/complicações , Cetose/epidemiologia , Cetose/imunologia , Masculino , Curva ROCRESUMO
Proteinuria predicts accelerated decline in kidney function in kidney transplant recipients (KTRs). We hypothesized that aberrant filtration of complement factors causes intraluminal activation, apical membrane attack on tubular cells, and progressive injury. Biobanked samples from two previous studies in albuminuric KTRs were used. The complement-activation split products C3c, C3dg, and soluble C5b-9-associated C9 neoantigen were analyzed by ELISA in urine and plasma using neoepitope-specific antibodies. Urinary extracellular vesicles (uEVs) were enriched by lectin and immunoaffinity isolation and analyzed by immunoblot analysis. Urine complement excretion increased significantly in KTRs with an albumin-to-creatinine ratio of ≥300 mg/g compared with <30 mg/g. Urine C3dg and C9 neoantigen excretion correlated significantly to changes in albumin excretion from 3 to 12 mo after transplantation. Fractional excretion of C9 neoantigen was significantly higher than for albumin, indicating postfiltration generation. C9 neoantigen was detected in uEVs in six of the nine albuminuric KTRs but was absent in non-albuminuric controls (n = 8). In C9 neoantigen-positive KTRs, lectin affinity enrichment of uEVs from the proximal tubules yielded signal for iC3b, C3dg, C9 neoantigen, and Na+-glucose transporter 2 but only weakly for aquaporin 2. Coisolation of podocyte markers and Tamm-Horsfall protein was minimal. Our findings show that albuminuria is associated with aberrant filtration and intratubular activation of complement with deposition of C3 activation split products and C5b-9-associated C9 neoantigen on uEVs from the proximal tubular apical membrane. Intratubular complement activation may contribute to progressive kidney injury in proteinuric kidney grafts.NEW & NOTEWORTHY The present study proposes a mechanistic coupling between proteinuria and aberrant filtration of complement precursors, intratubular complement activation, and apical membrane attack in kidney transplant recipients. C3dg and C5b-9-associated C9 neoantigen associate with proximal tubular apical membranes as demonstrated in urine extracellular vesicles. The discovery suggests intratubular complement as a mediator between proteinuria and progressive kidney damage. Inhibitors of soluble and/or luminal complement activation with access to the tubular lumen may be beneficial.
Assuntos
Albuminúria/imunologia , Membrana Celular/imunologia , Ativação do Complemento , Complemento C3b/urina , Complexo de Ataque à Membrana do Sistema Complemento/urina , Células Epiteliais/imunologia , Vesículas Extracelulares/imunologia , Transplante de Rim/efeitos adversos , Túbulos Renais Proximais/imunologia , Fragmentos de Peptídeos/urina , Adolescente , Adulto , Idoso , Albuminúria/sangue , Albuminúria/urina , Membrana Celular/metabolismo , Estudos Transversais , Células Epiteliais/metabolismo , Vesículas Extracelulares/metabolismo , Humanos , Túbulos Renais Proximais/metabolismo , Pessoa de Meia-Idade , Fragmentos de Peptídeos/sangue , Resultado do Tratamento , Adulto JovemRESUMO
Background: Diabetic kidney disease (DKD) is the most common cause of kidney failure in the world, and novel predictive biomarkers and molecular mechanisms of disease are needed. Endothelial cell-specific molecule-1 (Esm-1) is a secreted proteoglycan that attenuates inflammation. We previously identified that a glomerular deficiency of Esm-1 associates with more pronounced albuminuria and glomerular inflammation in DKD-susceptible relative to DKD-resistant mice, but its contribution to DKD remains unexplored. Methods: Using hydrodynamic tail-vein injection, we overexpress Esm-1 in DKD-susceptible DBA/2 mice and delete Esm-1 in DKD-resistant C57BL/6 mice to study the contribution of Esm-1 to DKD. We analyze clinical indices of DKD, leukocyte infiltration, podocytopenia, and extracellular matrix production. We also study transcriptomic changes to assess potential mechanisms of Esm-1 in glomeruli. Results: In DKD-susceptible mice, Esm-1 inversely correlates with albuminuria and glomerular leukocyte infiltration. We show that overexpression of Esm-1 reduces albuminuria and diabetes-induced podocyte injury, independent of changes in leukocyte infiltration. Using a complementary approach, we find that constitutive deletion of Esm-1 in DKD-resistant mice modestly increases the degree of diabetes-induced albuminuria versus wild-type controls. By glomerular RNAseq, we identify that Esm-1 attenuates expression of kidney disease-promoting and interferon (IFN)-related genes, including Ackr2 and Cxcl11. Conclusions: We demonstrate that, in DKD-susceptible mice, Esm-1 protects against diabetes-induced albuminuria and podocytopathy, possibly through select IFN signaling. Companion studies in patients with diabetes suggest a role of Esm-1 in human DKD.
Assuntos
Albuminúria , Diabetes Mellitus Experimental , Nefropatias Diabéticas , Células Endoteliais , Inflamação , Animais , Camundongos , Albuminúria/imunologia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/metabolismo , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/imunologia , Suscetibilidade a Doenças/metabolismo , Células Endoteliais/metabolismo , Inflamação/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Fatores de Transcrição/metabolismoRESUMO
IgA nephropathy (IgAN) is the most common type of glomerulonephritis worldwide, which follows a chronic but nonetheless highly variable course of progression. IgA immune complexes are the primary source of renal deposits in IgAN. Apart from the presence of granular IgA1 deposits in the glomerular mesangium and mesangial hypercellularity as common features, the detailed process of IgA1 deposition and clearance in the kidney remains unclear. We sought to examine the dynamics of IgA deposition and tissue plasticity in response to deposits including their intrarenal clearance. We followed a synthetic approach to produce a recombinant fusion between IgA Fc (rIgA) and a biotin tag, which was subsequently induced with streptavidin (SA) to form an oligomeric poly-IgA mimic. Both uninduced rIgA (mono-rIgA) and polymeric SA-rIgA (poly-rIgA) were injected intravenously into Wistar rats. Plasma IgA levels and renal and liver histology were examined in a time series. In contrast to mono-rIgA, this synthetic poly-rIgA analog formed renal deposits exclusively in the glomerulus and were mostly cleared in 3 h. However, repeated daily injections for 12 days caused long-lasting and stronger glomerular IgA deposition together with IgG and complement C3, in association with mesangial cell proliferation, matrix expansion, and variable degrees of albuminuria and hematuria that phenocopied IgAN. Ex vivo, poly-rIgA bound cultured mesangial cells and elicited cytokine production, in addition to activating plasma C3 that was consistent with the actions of IgA immune complexes in IgAN pathogenesis. Remarkably, the kidneys were able to reverse all pathologic manifestations and restore normal glomerular histology 2 weeks after injections were halted. The synthetic model showed the kinetics between the intricate balance of renal deposition and clearance, as well as glomerular plasticity towards healing. Together, the results revealed a priming effect of existing deposits in promoting stronger and longer-lasting IgA deposition to cause renal damage. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Assuntos
Complemento C3/imunologia , Glomerulonefrite por IGA/patologia , Glomerulonefrite/patologia , Imunoglobulina A/imunologia , Albuminúria/imunologia , Albuminúria/patologia , Animais , Modelos Animais de Doenças , Glomerulonefrite/imunologia , Glomerulonefrite por IGA/imunologia , Hematúria/imunologia , Hematúria/patologia , Humanos , Imunoglobulina G/imunologia , Rim/imunologia , Rim/patologia , Glomérulos Renais/imunologia , Glomérulos Renais/patologia , Masculino , Células Mesangiais/imunologia , Ratos , Ratos Wistar , Proteínas RecombinantesRESUMO
Hypertension and kidney involvement are common in patients with autoimmune disease. Sodium intake is linked to hypertension in both human and animal studies. Evidence suggests that dietary salt may be an important environmental factor that promotes autoimmune activity. Therefore, we hypothesized that a long-term high-salt diet would accelerate the progression of autoimmunity, hypertension, and albuminuria during systemic lupus erythematosus (SLE), an autoimmune disease that predominantly affects young women and has a high prevalence of hypertension and renal disease. To test this hypothesis, an established experimental model of SLE (female NZBWF1 mice) that develops hypertension and renal disease was used. SLE mice were fed a high-salt (4% NaCl) or normal (0.4% NaCl) diet for 24 wk beginning at 10 wk of age and ending at 34 wk of age, a time by which female NZBWF1 mice typically have hypertension and exhibit signs of renal disease. Plasma anti-dsDNA autoantibodies were measured as an indicator of active SLE disease, and urinary albumin was monitored longitudinally as a marker of renal disease. Arterial pressure was measured in conscious, freely moving mice at 34 wk of age. Urinary endothelin-1 (ET-1) excretion, renal endothelin A and B receptor protein expression, and renal mRNA expression of NOS1, NOS2, NOX2, MCP-1, TNF-α, serum- and glucocorticoid-regulated kinase 1, and interleukin-2 (IL-2) were assessed to determine the impact on gene products commonly altered by a high-salt diet. SLE mice fed a high-salt diet had increased circulating autoantibodies, but the high-salt diet did not significantly affect albuminuria or arterial pressure. Urinary ET-1 excretion was increased, whereas renal endothelin A receptor and IL-2 expression were decreased in response to a high-salt diet. These data suggest that a chronic high-salt diet may not accelerate cardiovascular and renal consequences commonly associated with SLE.
Assuntos
Albuminúria/fisiopatologia , Pressão Sanguínea/fisiologia , Rim/fisiopatologia , Lúpus Eritematoso Sistêmico/fisiopatologia , Sódio na Dieta , Albuminúria/imunologia , Animais , Autoanticorpos , DNA/imunologia , Modelos Animais de Doenças , Progressão da Doença , Feminino , Hipertensão/imunologia , Hipertensão/fisiopatologia , Rim/imunologia , Lúpus Eritematoso Sistêmico/imunologia , CamundongosRESUMO
BACKGROUND AND AIMS: Proteins containing advanced glycation end products are highly immunogenic and anti-advanced glycation end products antibodies (anti-AGEs antibodies) are found in the sera of diabetics. MATERIALS AND METHODS: Enzyme-linked immunosorbent assay (ELISA) was used for measuring levels of anti-advanced glycation end products antibodies in sera of 93 patients with type 2 diabetes mellitus and arterial hypertension (mean age 61.4±11.3 years, diabetes duration 9.88±3.12 years; hypertension duration 9.28±4.98). These values were compared to serum anti-AGEs antibodies in 42 age and sex matched controls. Diabetics were divided in two groups according to presence or absence of microangiopathy, group 1 (n=67) and group 2 (n=26), respectively. RESULTS: Serum levels of anti-AGEs antibodies in patients with type 2 diabetes mellitus and arterial hypertension were statistically significantly higher than those in the control group (1.39±0.39 vs. 1.05±0.32), (p<0.05). Group 1 showed significantly higher levels of anti-AGEs antibodies than those of healthy controls (1.53±0.14 vs. 1.05±0.32), (p<0.01). Anti-AGEs antibodies levels were higher in patients with microvascular complications than these in patients without complications. Anti-AGEs antibodies correlate with diastolic blood pressure (r=0.26, p=0.05) and body mass index (r=0.37, p=0.03). We found significantly higher percentage of positive patients for anti-AGEs antibodies (mean+2SD) in group 1 than in group 2. CONCLUSION: Determining the levels of serum anti-AGEs antibodies can help physicians make early diagnosis and prognosis of the severity of late diabetic complications in hypertensive patients.
Assuntos
Autoanticorpos/imunologia , Diabetes Mellitus Tipo 2/imunologia , Angiopatias Diabéticas/imunologia , Elastina/imunologia , Produtos Finais de Glicação Avançada/imunologia , Hipertensão/imunologia , Idoso , Albuminúria/etiologia , Albuminúria/imunologia , Estudos de Casos e Controles , Diabetes Mellitus Tipo 2/complicações , Angiopatias Diabéticas/etiologia , Neuropatias Diabéticas/etiologia , Neuropatias Diabéticas/imunologia , Retinopatia Diabética/etiologia , Retinopatia Diabética/imunologia , Feminino , Humanos , Hipertensão/complicações , Masculino , Pessoa de Meia-IdadeRESUMO
Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disease characterized by circulating autoantibodies, prevalent hypertension, renal injury, and cardiovascular disease. Onset of the disease often occurs in young women of childbearing age. Although kidney involvement is common to patients with SLE, little is known about temporal changes in renal hemodynamic function and its relationship to the pathogenesis of hypertension during autoimmune diseases. We hypothesized that the loss of immunological tolerance and subsequent production of autoantibodies in SLE leads to impaired renal hemodynamic function that precedes the development hypertension. Female NZBWF1 (SLE) mice and female NZW/LacJ (control) mice were instrumented with carotid artery and jugular vein catheters to determine mean arterial pressure (MAP) and glomerular filtration rate, respectively, at ages of 15, 20, 24, 28, 31, and 34 wk. In addition, urinary albumin excretion, blood urea nitrogen, circulating autoantibodies, and glomerulosclerosis were assessed at each age. Levels of circulating autoantibodies are increased between 24 and 28 wk of age in NZBWF1 mice and were significantly greater than in control mice. Glomerular filtration rate was significantly increased at 28 wk of age in NZBWF1 mice followed by a sharp decline at 34 wk of age. NZBWF1 mice had an increase in MAP that occurred by 34 wk of age. These data show that changes in circulating autoantibodies, renal hemodynamic function, and glomerular injury occur in NZBWF1 mice before changes in MAP, suggesting an important mechanistic role for autoimmunity to directly impair renal hemodynamic function and promote the development of hypertension.
Assuntos
Pressão Arterial , Hipertensão/fisiopatologia , Rim/fisiopatologia , Lúpus Eritematoso Sistêmico/fisiopatologia , Albuminúria/imunologia , Albuminúria/fisiopatologia , Animais , Autoanticorpos/sangue , Autoimunidade , Nitrogênio da Ureia Sanguínea , Modelos Animais de Doenças , Feminino , Taxa de Filtração Glomerular , Hipertensão/sangue , Hipertensão/imunologia , Rim/patologia , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/imunologia , Camundongos Endogâmicos NZB , Fatores de TempoRESUMO
Studies of Dahl salt-sensitive (SS) rats have shown that renal CD3+ T cells and ED-1+ macrophages are involved in the development of salt-sensitive hypertension and renal damage. The present study demonstrated that the increase in renal immune cells, which accompanies renal hypertrophy and albuminuria in high-salt diet-fed Dahl SS rats, is absent in Sprague-Dawley and SSBN13 rats that are protected from the SS disease phenotype. Flow cytometric analysis demonstrated that >70% of the immune cells in the SS kidney are M1 macrophages. PCR profiling of renal myeloid cells showed a salt-induced upregulation in 9 of 84 genes related to Toll-like receptor signaling, with notable upregulation of the Toll-like receptor 4/CD14/MD2 complex. Because of the prominent increase in macrophages in the SS kidney, we used liposome-encapsulated clodronate (Clod) to deplete macrophages and assess their contribution to salt-sensitive hypertension and renal damage. Dahl SS animals were administered either Clod-containing liposomes (Clod-Lipo), Clod, or PBS-containing liposomes as a vehicle control. Clod-Lipo treatment depleted circulating and splenic macrophages by â¼50%; however, contrary to our hypothesis, Clod-Lipo-treated animals developed an exacerbated salt-sensitive response with respect to blood pressure and albuminuria, which was accompanied by increased renal T and B cells. Interestingly, those treated with Clod also demonstrated an exacerbated phenotype, but it was less severe than Clod-Lipo-treated animals and independent of changes to the number of renal immune cells. Here, we have shown that renal macrophages in Dahl SS animals sustain a M1 proinflammatory phenotype in response to increased dietary salt and highlighted potential adverse effects of Clod-Lipo macrophage depletion.
Assuntos
Albuminúria/imunologia , Hipertensão/imunologia , Nefropatias/imunologia , Rim/imunologia , Macrófagos/imunologia , Cloreto de Sódio na Dieta , Albuminúria/etiologia , Albuminúria/metabolismo , Albuminúria/patologia , Animais , Linfócitos B/imunologia , Linfócitos B/metabolismo , Ácido Clodrônico/toxicidade , Modelos Animais de Doenças , Progressão da Doença , Hipertensão/etiologia , Hipertensão/metabolismo , Hipertensão/patologia , Rim/efeitos dos fármacos , Rim/metabolismo , Rim/patologia , Nefropatias/etiologia , Nefropatias/metabolismo , Nefropatias/patologia , Receptores de Lipopolissacarídeos/metabolismo , Antígeno 96 de Linfócito/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Fenótipo , Ratos Endogâmicos BN , Ratos Endogâmicos Dahl , Ratos Sprague-Dawley , Transdução de Sinais , Linfócitos T/imunologia , Linfócitos T/metabolismo , Receptor 4 Toll-Like/metabolismoRESUMO
Diabetic nephropathy (DN) is one of the most common complications of diabetes, and currently the first end-stage renal disease worldwide. New strategies to treat DN using agents that target inflammatory pathways have attracted special interest. Recent pieces of evidences suggest a promising effect of IL-17A, the Th17 effector cytokine. Among experimental DN models, mouse strain BTBR ob/ob (leptin deficiency mutation) develops histological features similar to human DN, which means an opportunity to study mechanisms and novel therapies aimed at DN regression. We found that BTBR ob/ob mice presented renal activation of the factors controlling Th17 differentiation. The presence of IL-17A-expressing cells, mainly CD4+ and γδ lymphocytes, was associated with upregulation of proinflammatory factors, macrophage infiltration and the beginning of renal damage. To study IL-17A involvement in experimental DN pathogenesis, treatment with an IL-17A neutralizing antibody was carried out starting when the renal damage had already appeared. IL-17A blockade ameliorated renal dysfunction and disease progression in BTBR ob/ob mice. These beneficial effects correlated to podocyte number restoration and inhibition of NF-κB/proinflammatory factors linked to a decrease in renal inflammatory-cell infiltration. These data demonstrate that IL-17A takes part in diabetes-mediated renal damage and could be a promising therapeutic target to improve DN.
Assuntos
Albuminúria/tratamento farmacológico , Anticorpos Neutralizantes/administração & dosagem , Nefropatias Diabéticas/tratamento farmacológico , Interleucina-17/antagonistas & inibidores , Albuminúria/genética , Albuminúria/imunologia , Albuminúria/patologia , Animais , Nefropatias Diabéticas/genética , Nefropatias Diabéticas/imunologia , Nefropatias Diabéticas/urina , Progressão da Doença , Humanos , Interleucina-17/imunologia , Interleucina-17/metabolismo , Rim/efeitos dos fármacos , Rim/imunologia , Rim/patologia , Leptina/genética , Masculino , Camundongos , Camundongos Transgênicos , Células Th17/efeitos dos fármacos , Células Th17/imunologia , Células Th17/metabolismoRESUMO
BACKGROUND: Kidney tubulointerstitial fibrosis on biopsy is a strong predictor of chronic kidney disease (CKD) progression, and CKD is associated with elevated risk of cardiovascular disease (CVD). Tubular health is poorly quantified by traditional kidney function measures, including estimated glomerular filtration rate (eGFR) and albuminuria. We hypothesized that urinary biomarkers of tubular injury, inflammation, and repair would be associated with higher risk of CVD and mortality in persons with CKD. METHODS: We measured urinary concentrations of interleukin-18 (IL-18), kidney injury molecule-1, neutrophil gelatinase-associated lipocalin, monocyte chemoattractant protein-1, and chitinase-3-like protein-1 (YKL-40) at baseline among 2,377 participants of the Systolic Blood Pressure Intervention Trial who had an eGFR < 60 mL/min/1.73 m2. We used Cox proportional hazards models to evaluate biomarker associations with CVD events and all-cause mortality. RESULTS: At baseline, the mean age of participants was 72 ± 9 years, and eGFR was 48 ± 11 mL/min/1.73 m2. Over a median follow-up of 3.8 years, 305 CVD events (3.6% per year) and 233 all-cause deaths (2.6% per year) occurred. After multivariable adjustment including eGFR, albuminuria, and urinary creatinine, none of the biomarkers showed statistically significant associations with CVD risk. Urinary IL-18 (hazard ratio [HR] per 2-fold higher value, 1.14; 95% CI 1.01-1.29) and YKL-40 (HR per 2-fold higher value, 1.08; 95% CI 1.02-1.14) concentrations were each incrementally associated with higher mortality risk. Associations were similar when stratified by randomized blood pressure arm. CONCLUSIONS: Among hypertensive trial participants with CKD, higher urinary IL-18 and YKL-40 were associated with higher risk of mortality, but not CVD.
Assuntos
Albuminúria/diagnóstico , Doenças Cardiovasculares/epidemiologia , Hipertensão/tratamento farmacológico , Túbulos Renais/patologia , Insuficiência Renal Crônica/mortalidade , Idoso , Idoso de 80 Anos ou mais , Albuminúria/imunologia , Albuminúria/patologia , Albuminúria/urina , Anti-Hipertensivos/administração & dosagem , Biomarcadores/urina , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Determinação da Pressão Arterial/normas , Doenças Cardiovasculares/etiologia , Progressão da Doença , Feminino , Fibrose , Taxa de Filtração Glomerular/fisiologia , Humanos , Hipertensão/fisiopatologia , Hipertensão/urina , Túbulos Renais/imunologia , Masculino , Prognóstico , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/imunologia , Insuficiência Renal Crônica/urinaRESUMO
De novo expression of CD44 in glomerular parietal epithelial cells (PECs) leads to a prosclerotic and migratory PEC phenotype in glomerulosclerosis. However, the regulatory mechanisms underlying CD44 expression by activated PECs remain largely unknown. This study was performed to examine the mediators responsible for CD44 induction in glomerular PECs in association with diabetes. CD44 expression and localization were evaluated in the glomeruli of Zucker diabetic rat kidneys and primary cultured PECs upon albumin stimulation. Real-time polymerase chain reaction confirmed an albuminuria-associated upregulation of the CD44 gene in the glomeruli of diabetic rats. Immunostaining analysis of diabetic kidneys further revealed an increase in CD44 in hypertrophic PECs, which often contain albumin-positive vesicles. Losartan treatment significantly attenuated albuminuria and lowered CD44 protein levels in the diabetic kidneys. In primary cultured rat PECs, rat serum albumin (0.25-1 mg/ml) caused a dose-dependent upregulation of CD44, claudin-1, and megalin protein expression, which was accompanied by an activation of extracellular signal-regulated kinase1/2 (ERK1/2) signaling. Albumin-induced CD44 and claudin-1 expression were greatly suppressed in the presence of the ERK1/2 inhibitor, U0126. In addition, knockdown of megalin by small interfering RNA interference in PECs resulted in a significant reduction of albumin-induced CD44 and claudin-1 proteins. Taken together, our results demonstrate that albumin induces CD44 expression by PECs via the activation of the ERK signaling pathway, which is partially mediated by endocytic receptor megalin.
Assuntos
Albuminúria/enzimologia , Nefropatias Diabéticas/enzimologia , Células Epiteliais/efeitos dos fármacos , Receptores de Hialuronatos/metabolismo , Glomérulos Renais/efeitos dos fármacos , Proteína Quinase 1 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Albumina Sérica/farmacologia , Albuminúria/imunologia , Albuminúria/patologia , Animais , Células Cultivadas , Claudina-1/metabolismo , Nefropatias Diabéticas/imunologia , Nefropatias Diabéticas/patologia , Modelos Animais de Doenças , Endocitose , Ativação Enzimática , Células Epiteliais/enzimologia , Células Epiteliais/imunologia , Células Epiteliais/patologia , Receptores de Hialuronatos/genética , Glomérulos Renais/enzimologia , Glomérulos Renais/imunologia , Glomérulos Renais/patologia , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Masculino , Ratos Sprague-Dawley , Ratos Zucker , Reabsorção Renal , Transdução de Sinais , Regulação para CimaRESUMO
The quantification of urine albumin is a common practice in Medical Biology laboratories. It allows the assessment of renal injury in common pathologies and many studies have confirmed its role in the diagnosis and prognosis of these disorders. The physicochemical characteristics of albumin in the urine, very different from those in the blood, do not allow the use of the same standardized assay techniques for the blood albumin determination and make it difficult its quantification. Indeed, because of a physiological fragmentation phenomenon, urinary albumin is present in the urine as various small specific peptides. We will present here the main methods of determination of albumin in the urine, which are immuno-turbidimetric and immuno-nephelometric methods, high performance liquid chromatography with steric exclusion and liquid chromatography coupled with mass spectrometry. Currently, immunoanalysis techniques are the most used and are not standardized; large bias can be found between the different kits. This observation calls for a standardization of its determination in the urine.
Assuntos
Albuminúria/diagnóstico , Albuminúria/urina , Urinálise/métodos , Albuminúria/imunologia , Cromatografia Líquida de Alta Pressão/normas , Cromatografia Líquida , Humanos , Imunoensaio , Nefelometria e Turbidimetria/métodos , Nefelometria e Turbidimetria/normas , Padrões de Referência , Albumina Sérica/análise , Albumina Sérica/imunologia , Espectrometria de Massas em Tandem/normas , Urinálise/normas , Coleta de Urina/métodos , Coleta de Urina/normasRESUMO
Diabetic Nephropathy (DN) is an important cause of morbidity and death amongst diabetes. Persistent micro and macroalbuminuria are well known predictors of DN leading to progressive end-stage renal disease. However, albuminuria has several limitations. Increasing evidences show that YKL-40 is highly expressed in variety of inflammatory diseases and also recognized as a non-invasive prognostic biomarker for inflammation. In the present study, we measured plasma YKL-40 levels in different stages of albuminuria and assessed its diagnostic accuracy as a biomarker for DN and correlated with different families of circulatory cytokines. A total of 306 subjects were recruited and divided into three groups [Group-I, control (n = 83), Group-II, Normoalbuminuria (n = 81), Group-III, DN (n = 142)]. Group-III is further subdivided into: Group-IIIa, microalbuminuria (n = 73), Group-IIIb, macroalbuminuria (n = 69). The median levels of YKL-40 (p = 0.001) showed a marked stepwise increase from normo to macroalbuminuria and positively correlated with eGFR. The AUCROC for YKL-40 was found to be high [0.95; (95% CI: 0.88-1.0)], when compared to other acute phase markers. Plasma YKL-40 showed a positive correlation with LIGHT/TNFSF14, sIL-6Ra, gp130/sIL-6Rß, IFN-ß, IL-8, TNFSF14, sCD-30 and eGFR meanwhile a negative correlation with TWEAK/TNFSF12, IL-7 like cytokine and IFN-λ2. Plasma YKL-40 could be a potential biomarker for early diagnosis of incipient DN among South Indian population.
Assuntos
Albuminúria/diagnóstico , Biomarcadores/sangue , Proteína 1 Semelhante à Quitinase-3/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Nefropatias Diabéticas/diagnóstico , Adulto , Idoso , Albuminúria/imunologia , Citocinas/metabolismo , Diabetes Mellitus Tipo 2/imunologia , Nefropatias Diabéticas/imunologia , Progressão da Doença , Diagnóstico Precoce , Feminino , Humanos , Índia , Mediadores da Inflamação/metabolismo , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
Immunoglobulin A (IgA) nephropathy (IgAN), the most common glomerulonephritis worldwide, is characterized by IgA depositions in the kidney. Deficiency of CD37, a leukocyte-specific tetraspanin, leads to spontaneous development of renal pathology resembling IgAN. However, the underlying molecular mechanism has not been resolved. Here we found that CD37 expression on B cells of patients with IgAN was significantly decreased compared to B cells of healthy donors. Circulating interleukin (IL)-6 levels, but not tumor necrosis factor-α or IL-10, were elevated in Cd37-/- mice compared to wild-type mice after lipopolysaccharide treatment. Cd37-/- mice displayed increased glomerular neutrophil influx, immune complex deposition, and worse renal function. To evaluate the role of IL-6 in the pathogenesis of accelerated renal pathology in Cd37-/-mice, we generated Cd37xIl6 double-knockout mice. These double-knockout and Il6-/- mice displayed no glomerular IgA deposition and were protected from exacerbated renal failure following lipopolysaccharide treatment. Moreover, kidneys of Cd37-/- mice showed more mesangial proliferation, endothelial cell activation, podocyte activation, and segmental podocyte foot process effacement compared to the double-knockout mice, emphasizing that IL-6 mediates renal pathology in Cd37-/- mice. Thus, our study indicates that CD37 may protect against IgA nephropathy by inhibition of the IL-6 pathway.
Assuntos
Glomerulonefrite por IGA/metabolismo , Imunoglobulina A/metabolismo , Interleucina-6/metabolismo , Glomérulos Renais/metabolismo , Tetraspaninas/deficiência , Albuminúria/imunologia , Albuminúria/metabolismo , Albuminúria/prevenção & controle , Animais , Antígenos CD/genética , Antígenos de Neoplasias/sangue , Antígenos de Neoplasias/genética , Linfócitos B/imunologia , Linfócitos B/metabolismo , Estudos de Casos e Controles , Proliferação de Células , Modelos Animais de Doenças , Predisposição Genética para Doença , Glomerulonefrite por IGA/imunologia , Glomerulonefrite por IGA/patologia , Glomerulonefrite por IGA/prevenção & controle , Humanos , Imunoglobulina A/imunologia , Interleucina-6/deficiência , Interleucina-6/genética , Glomérulos Renais/imunologia , Glomérulos Renais/patologia , Glomérulos Renais/fisiopatologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infiltração de Neutrófilos , Fenótipo , Podócitos/imunologia , Podócitos/metabolismo , Podócitos/patologia , Tetraspaninas/sangue , Tetraspaninas/genéticaRESUMO
The present study, performed in Dahl salt-sensitive (SS) and SS- Rag1-/- rats lacking T and B lymphocytes, tested the hypothesis that immune cells amplify salt-sensitive hypertension and kidney damage in response to a high-protein diet. After being weaned, SS and SS- Rag1-/- rats were placed on an isocaloric, 0.4% NaCl diet containing normal (18%) or high (30%) protein. At 9 wk of age, rats were switched to a 4.0% NaCl diet containing the same amount of dietary protein and maintained on the high-salt diet for 3 wk. After being fed the high-salt diet, SS rats fed high protein had amplified hypertension and albumin excretion (158.7 ± 2.6 mmHg and 140.8 ± 16.0 mg/day, respectively, means ± SE) compared with SS rats fed normal protein (139.4 ± 3.6 mmHg and 69.4 ± 11.3 mg/day). When compared with the SS rats, SS- Rag1-/- rats fed high protein were protected from exacerbated hypertension and albuminuria (142.9 ± 5.8 mmHg and 66.2 ± 10.8 mg/day). After 3 wk of the high-salt diet, there was a corresponding increase in total leukocyte infiltration (CD45+) in the kidneys of both strains fed high-protein diet. The SS- Rag1-/- rats fed high-protein diet had 74-86% fewer CD3+ T lymphocytes and CD45R+ B lymphocytes infiltrating the kidney versus SS rats, but there was no difference in the infiltration of CD11b/c+ monocytes and macrophages, suggesting that the protective effects observed in the SS- Rag1-/- rats are specific to the reduction of lymphocytes. With the SS- Rag1-/- rats utilized as a novel tool to explore the effects of lymphocyte deficiency, these results provide evidence that adaptive immune mechanisms contribute to the exacerbation of salt-induced hypertension and renal injury mediated by increased dietary protein intake.
Assuntos
Imunidade Adaptativa , Linfócitos B/imunologia , Pressão Sanguínea , Dieta Rica em Proteínas/efeitos adversos , Genes RAG-1 , Hipertensão/imunologia , Nefropatias/imunologia , Rim/imunologia , Cloreto de Sódio na Dieta , Linfócitos T/imunologia , Albuminúria/genética , Albuminúria/imunologia , Albuminúria/fisiopatologia , Animais , Linfócitos B/metabolismo , Complexo CD3/deficiência , Complexo CD3/genética , Modelos Animais de Doenças , Hipertensão/sangue , Hipertensão/genética , Hipertensão/fisiopatologia , Rim/metabolismo , Rim/fisiopatologia , Nefropatias/sangue , Nefropatias/genética , Nefropatias/fisiopatologia , Masculino , Ratos Endogâmicos Dahl , Ratos Transgênicos , Fatores de Risco , Linfócitos T/metabolismoRESUMO
Soluble cytokine receptors may play an important role in development of microalbuminuria (MA) in type-1 diabetes (T1D). In this study, we measured 12 soluble receptors and ligands from TNF-α/IL6/IL2 pathways in T1D patients with MA (n = 89) and T1D patients without MA (n = 483) participating in the PAGODA study. Twelve proteins in the sera from T1D patients with and without MA were measured using multiplex Luminex assays. Ten serum proteins (sTNFR1, sTNFR2, sIL2Rα, MMP2, sgp130, sVCAM1, sIL6R, SAA, CRP, and sICAM1) were significantly elevated in T1D patients with MA. After adjusting for age, duration of diabetes, and sex in logistic regression, association remained significant for seven proteins. MA is associated with increasing concentrations of all 10 proteins, with the strongest associations observed for sTNFR1 (OR = 108.3, P < 10-32) and sTNFR2 (OR = 65.5, P < 10-37), followed by sIL2Rα (OR = 12.9, P < 10-13), MMP2 (OR = 5.5, P < 10-6), sgp130 (OR = 5.2, P < 10-3), sIL6R (OR = 4.6, P < 10-4), and sVCAM1 (OR = 3.3, P < 10-4). We developed a risk score system based on the combined odds ratios associated with each quintile for each protein. The risk scores cluster MA patients into three subsets, each associated with distinct risk for MA attributable to proteins in the TNF-α/IL6 pathway (mean OR = 1, 13.5, and 126.3 for the three subsets, respectively). Our results suggest that the TNF-α/IL6 pathway is overactive in approximately 40% of the MA patients and moderately elevated in the middle 40% of the MA patients. Our results suggest the existence of distinct subsets of MA patients identifiable by their serum protein profiles.
Assuntos
Albuminúria/imunologia , Diabetes Mellitus Tipo 1/sangue , Interleucina-6/sangue , Fator de Necrose Tumoral alfa/sangue , Adulto , Idoso , Albuminúria/sangue , Biomarcadores/sangue , Proteínas Sanguíneas/análise , Diabetes Mellitus Tipo 1/complicações , Feminino , Humanos , Inflamação , Masculino , Pessoa de Meia-Idade , Razão de Chances , Receptores de Citocinas/sangue , Receptores Tipo I de Fatores de Necrose Tumoral/sangue , Receptores Tipo II do Fator de Necrose Tumoral/sangue , Adulto JovemRESUMO
To study the features of secretion of melatonin in the urine in patients with DM type 2 and NAFLD with manifestations of fibrosis and its relationship with some metabolic and immunological parameters, 23 patients with DM type 2 and NAFLD were examined. The degree of fibrosis in patients was diagnosed on the basis of static elastography and the study of indirect fibrosis markers 16 persons (72%) diagnosed with mild fibrosis (F0-F1 on METAVIR), 5 people (18.2%) - with moderate fibrosis (F2-F3 on METAVIR). Only 2 (8.7%) patients did not have any fibrotic disorders, so they were excluded from the further study. All patients underwent determination of melatonin excretion of albumin and in daily urine, as well as the determination of homocysteine in the blood. The level of excretion of melatonin in the urine in patients with DM type 2 and NAFLD did not depend on the degree of fibrosis and on the average was 89.50±16.66 mmol/day, which exceeded the reference values. It has been established that the increase in melatonin level in patients with DM type 2 and NAFLD is associated with the presence of fibrotic changes in the liver and a decrease in the activity of the inflammatory process. In addition, a direct correlation was found between the excretion of melatonin and homocysteine (r=0.43), as well as between melatonin and albumin excretion in the urine (r=0.20). Thus, an increased level of excretion of melatonin in the urine can be not only a marker of liver fibrosis, but also a predictor of cardiovascular disorders in patients with DM type 2 and NAFLD.
Assuntos
Albuminúria/diagnóstico , Diabetes Mellitus Tipo 2/diagnóstico , Cirrose Hepática/diagnóstico , Melatonina/urina , Hepatopatia Gordurosa não Alcoólica/diagnóstico , Adolescente , Adulto , Albuminúria/sangue , Albuminúria/imunologia , Albuminúria/urina , Biomarcadores/sangue , Biomarcadores/urina , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/patologia , Estudos de Coortes , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/imunologia , Diabetes Mellitus Tipo 2/urina , Técnicas de Imagem por Elasticidade , Feminino , Homocisteína/sangue , Homocisteína/imunologia , Humanos , Cirrose Hepática/sangue , Cirrose Hepática/imunologia , Cirrose Hepática/urina , Masculino , Melatonina/imunologia , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/sangue , Hepatopatia Gordurosa não Alcoólica/imunologia , Hepatopatia Gordurosa não Alcoólica/urinaRESUMO
The present study assessed the importance of immunity in angiotensin (ANG) II (5 ng·kg-1·min-1 iv)-mediated hypertension in Dahl salt-sensitive (SS) rats and SS rats deficient in T and B lymphocytes (SSRag1-/-) fed a 0.4% NaCl diet. Baseline mean arterial blood pressure (MAP) was not different between groups. ANG II infusion significantly increased MAP in both groups, although MAP increased more rapidly in SS rats, and the maximal MAP achieved was significantly greater in SS than SSRag1-/- rats (190 ± 3 vs. 177 ± 3 mmHg) after 12 days. Renal damage, as assessed by albumin excretion rate, was significantly increased after 12 days of ANG lI infusion in SS (from 32 ± 4 to 81 ± 9 mg/day) and SSRag1-/- (from 12 ± 2 to 51 ± 8 mg/day) rats; albumin excretion rate was significantly different between SS and SSRag1-/- rats at all points measured. After 9 days of recovery from ANG II, MAP was decreased to a greater extent in SSRag1-/- than SS rats (143 ± 5 vs. 157 ± 8 mmHg) compared with the peak MAP during ANG II infusion. At this same time point, albumin excretion rate was significantly lower in SSRag1-/- than SS rats (42 ± 8 vs. 66 ± 7 mg/day). Further studies demonstrated an increase in CD45+ total leukocytes, CD11b/c+ macrophages/monocytes, and CD3+ T cells in kidneys of ANG II- compared with vehicle-treated SS rats. The present data suggest that infiltrating T cells in the kidney exacerbate renal damage in ANG II-induced hypertension in SS rats maintained on a 0.4% NaCl diet, similar to results observed with a salt stimulus in SS rats.
Assuntos
Angiotensina II , Hipertensão/imunologia , Nefropatias/imunologia , Rim/imunologia , Linfócitos T/imunologia , Albuminúria/induzido quimicamente , Albuminúria/imunologia , Albuminúria/metabolismo , Albuminúria/fisiopatologia , Animais , Pressão Arterial , Linfócitos B/imunologia , Linfócitos B/metabolismo , Quimiotaxia de Leucócito , Citocinas/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Genes RAG-1 , Hipertensão/induzido quimicamente , Hipertensão/metabolismo , Hipertensão/fisiopatologia , Rim/metabolismo , Rim/patologia , Rim/fisiopatologia , Nefropatias/induzido quimicamente , Nefropatias/metabolismo , Nefropatias/fisiopatologia , Masculino , Fenótipo , Ratos Endogâmicos Dahl , Ratos Sprague-Dawley , Ratos Transgênicos , Cloreto de Sódio na Dieta , Linfócitos T/metabolismoRESUMO
A key feature of glomerular diseases such as crescentic glomerulonephritis and focal segmental glomerulosclerosis is the activation, migration and proliferation of parietal epithelial cells. CD44-positive activated parietal epithelial cells have been identified in proliferative cellular lesions in glomerular disease. However, it remains unknown whether CD44-positive parietal epithelial cells contribute to the pathogenesis of scarring glomerular diseases. Here, we evaluated this in experimental crescentic glomerulonephritis and the transgenic anti-Thy1.1 model for collapsing focal segmental glomerulosclerosis in CD44-deficient (cd44-/-) and wild type mice. For both models albuminuria was significantly lower in cd44-/- compared to wild type mice. The number of glomerular Ki67-positive proliferating cells was significantly reduced in cd44-/- compared to wild type mice, which was associated with a reduced number of glomerular lesions in crescentic glomerulonephritis. In collapsing focal segmental glomerulosclerosis, the extracapillary proliferative cellular lesions were smaller in cd44-/- mice, but the number of glomerular lesions was not different compared to wild type mice. For crescentic glomerulonephritis the influx of granulocytes and macrophages into the glomerulus was similar. In vitro, the growth of CD44-deficient murine parietal epithelial cells was reduced compared to wild type parietal epithelial cells, and human parietal epithelial cell migration could be inhibited using antibodies directed against CD44. Thus, CD44-positive proliferating glomerular cells, most likely parietal epithelial cells, are essential in the pathogenesis of scarring glomerular disease.
Assuntos
Doença Antimembrana Basal Glomerular/imunologia , Células Epiteliais/imunologia , Glomerulosclerose Segmentar e Focal/imunologia , Receptores de Hialuronatos/imunologia , Glomérulos Renais/imunologia , Albuminúria/genética , Albuminúria/imunologia , Albuminúria/metabolismo , Animais , Doença Antimembrana Basal Glomerular/genética , Doença Antimembrana Basal Glomerular/metabolismo , Doença Antimembrana Basal Glomerular/patologia , Autoanticorpos/imunologia , Movimento Celular , Proliferação de Células , Células Cultivadas , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Proteínas da Matriz Extracelular/metabolismo , Predisposição Genética para Doença , Glomerulosclerose Segmentar e Focal/genética , Glomerulosclerose Segmentar e Focal/metabolismo , Glomerulosclerose Segmentar e Focal/patologia , Granulócitos/imunologia , Granulócitos/metabolismo , Receptores de Hialuronatos/genética , Receptores de Hialuronatos/metabolismo , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Transdução de Sinais , Antígenos Thy-1/genética , Antígenos Thy-1/imunologia , Antígenos Thy-1/metabolismoRESUMO
OBJECTIVES: The relationship between the renin-angiotensin system (RAS) and diabetes has been studied for many years. However, studies that assessed RAS components comprehensively were limited. We hypothesized that serum RAS components, especially the effector peptide angiotensin-II, might be closely associated with glucose metabolism status and diabetic complications. METHODS: We investigated the association of individual RAS component with albuminuria in 407 subjects with normal glucose metabolism (NGM), prediabetes, or type 2 diabetes mellitus (T2DM). Anthropometric and biochemical parameters, including glucose homeostasis, albuminuria, and RAS-related parameters such as plasma renin activity (PRA), aldosterone, angiotensin-converting enzyme (ACE), and angiotensin-II levels, were measured. RESULTS: The mean±standard deviation (SD) age and body mass index were 57.1±11.1years and 24.7±3.3kg/m2, respectively. There were 54 subjects with NGM, 102 with prediabetes, and 251 with T2DM. The mean±SD angiotensin-II levels in these groups were 9.32±6.89, 12.89±10.39, and 17.00±15.28pg/mL, and the respective urinary albumin-to-creatinine ratios (ACRs) were 8.1±5.3, 13.3±17.3, and 30.7±51.9mg/g, which were significantly different among the groups. The serum angiotensin-II levels were correlated with levels of PRA, insulin resistance, C-reactive protein, and urinary ACR. Among RAS-related parameters, only the angiotensin-II level was significantly associated with urinary ACR after adjusting for relevant risk factors. CONCLUSIONS: Angiotensin-II may play an important role in the development of albuminuria, particularly in subjects with impaired glucose metabolism.