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1.
FASEB J ; 35(1): e21213, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33368614

RESUMO

Preclinical studies have demonstrated that activation of the NOTCH pathway plays a key role in the pathogenesis of kidney damage. There is currently no information on the role of the Delta-like homologue 1 (DLK1), a NOTCH inhibitor, in the regulation of renal damage. Here, we investigated the contribution of DLK1 to experimental renal damage and the underlying molecular mechanisms. Using a Dlk1-null mouse model in the experimental renal damage of unilateral ureteral obstruction, we found activation of NOTCH, as shown by increased nuclear translocation of the NOTCH1 intracellular domain, and upregulation of Dlk2/hey-1 expression compared to wild-type (WT) littermates. NOTCH1 over-activation in Dlk1-null injured kidneys was associated with a higher inflammatory response, characterized by infiltration of inflammatory cells, mainly CD4/IL17A + lymphocytes, and activation of the Th17 immune response. Furthermore, pharmacological NOTCH blockade inhibited the transcription factors controlling Th17 differentiation and gene expression of the Th17 effector cytokine IL-17A and other related-inflammatory factors, linked to a diminution of inflammation in the injured kidneys. We propose that the non-canonical NOTCH ligand DLK1 acts as a NOTCH antagonist in renal injury regulating the Th17-mediated inflammatory response.


Assuntos
Proteínas de Ligação ao Cálcio/deficiência , Deleção de Genes , Imunidade Celular , Nefropatias/imunologia , Rim/imunologia , Células Th17/imunologia , Animais , Proteínas de Ligação ao Cálcio/imunologia , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/imunologia , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Rim/patologia , Nefropatias/genética , Nefropatias/patologia , Camundongos , Células Th17/patologia , Obstrução Ureteral/genética , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
2.
J Am Soc Nephrol ; 31(5): 983-995, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32209589

RESUMO

BACKGROUND: Expression of SerpinB2, a regulator of inflammatory processes, has been described in the context of macrophage activation and cellular senescence. Given that mechanisms for these processes interact and can shape kidney disease, it seems plausible that SerpinB2 might play a role in renal aging, injury, and repair. METHODS: We subjected SerpinB2 knockout mice to ischemia-reperfusion injury or unilateral ureteral obstruction. We performed phagocyte depletion to study SerpinB2's role beyond the effects of macrophages and transplanted bone marrow from knockout mice to wild-type mice and vice versa to dissect cell type-dependent effects. Primary tubular cells and macrophages from SerpinB2 knockout and wild-type mice were used for functional studies and transcriptional profiling. RESULTS: Cultured senescent tubular cells, kidneys of aged mice, and renal stress models exhibited upregulation of SerpinB2 expression. Functionally, lack of SerpinB2 in aged knockout mice had no effect on the magnitude of senescence markers but associated with enhanced kidney damage and fibrosis. In stress models, inflammatory cell infiltration was initially lower in knockout mice but later increased, leading to an accumulation of significantly more macrophages. SerpinB2 knockout tubular cells showed significantly reduced expression of the chemokine CCL2. Macrophages from knockout mice exhibited reduced phagocytosis and enhanced migration. Macrophage depletion and bone marrow transplantation experiments validated the functional relevance of these cell type-specific functions of SerpinB2. CONCLUSIONS: SerpinB2 influences tubule-macrophage crosstalk by supporting tubular CCL2 expression and regulating macrophage phagocytosis and migration. In mice, SerpinB2 expression seems to be needed for coordination and timely resolution of inflammation, successful repair, and kidney homeostasis during aging. Implications of SerpinB2 in human kidney disease deserve further exploration.


Assuntos
Injúria Renal Aguda/enzimologia , Envelhecimento/imunologia , Senescência Celular/imunologia , Túbulos Renais/enzimologia , Rim/enzimologia , Macrófagos/fisiologia , Inibidor 2 de Ativador de Plasminogênio/fisiologia , Traumatismo por Reperfusão/enzimologia , Obstrução Ureteral/complicações , Injúria Renal Aguda/etiologia , Injúria Renal Aguda/imunologia , Animais , Movimento Celular , Quimiocina CCL2/biossíntese , Quimiocina CCL2/genética , Técnicas de Cocultura , Indução Enzimática , Células Epiteliais/metabolismo , Fibrose , Homeostase , Rim/irrigação sanguínea , Rim/patologia , Masculino , Camundongos , Camundongos Knockout , Fagocitose , Inibidor 2 de Ativador de Plasminogênio/deficiência , Traumatismo por Reperfusão/imunologia , Transcriptoma , Obstrução Ureteral/enzimologia , Obstrução Ureteral/imunologia
3.
Clin Exp Immunol ; 199(1): 97-108, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31509227

RESUMO

Kidney injury significantly increases overall mortality. Neutrophilic granulocytes (neutrophils) are the most abundant human blood leukocytes. They are characterized by a high turnover rate, chiefly controlled by granulocyte colony stimulating factor (G-CSF). The role of kidney injury and uremia in regulation of granulopoiesis has not been reported. Kidney transplantation, which inherently causes ischemia-reperfusion injury of the graft, elevated human neutrophil expression of the surface glycoprotein CD177. CD177 is among the most G-CSF-responsive neutrophil genes and reversibly increased on neutrophils of healthy donors who received recombinant G-CSF. In kidney graft recipients, a transient rise in neutrophil CD177 correlated with renal tubular epithelial G-CSF expression. In contrast, CD177 was unaltered in patients with chronic renal impairment and independent of renal replacement therapy. Under controlled conditions of experimental ischemia-reperfusion and unilateral ureteral obstruction injuries in mice, renal G-CSF mRNA and protein expression significantly increased and systemic neutrophilia developed. Human renal tubular epithelial cell G-CSF expression was promoted by hypoxia and proinflammatory cytokine interleukin 17A in vitro. Clinically, recipients of ABO blood group-incompatible kidney grafts developed a larger rise in neutrophil CD177. Their grafts are characterized by complement C4d deposition on the renal endothelium, even in the absence of rejection. Indeed, complement activation, but not hypoxia, induced primary human endothelial cell G-CSF expression. Our data demonstrate that kidney injury induces renal G-CSF expression and modulates granulopoiesis. They delineate differential G-CSF regulation in renal epithelium and endothelium. Altered granulopoiesis may contribute to the systemic impact of kidney injury.


Assuntos
Basigina/metabolismo , Endotélio/metabolismo , Regulação da Expressão Gênica , Fator Estimulador de Colônias de Granulócitos/biossíntese , Neutrófilos/metabolismo , Insuficiência Renal/metabolismo , Trombopoese , Animais , Basigina/imunologia , Modelos Animais de Doenças , Endotélio/imunologia , Endotélio/patologia , Feminino , Fator Estimulador de Colônias de Granulócitos/imunologia , Humanos , Transplante de Rim , Masculino , Camundongos , Neutrófilos/imunologia , Neutrófilos/patologia , Insuficiência Renal/imunologia , Insuficiência Renal/patologia , Insuficiência Renal/cirurgia , Traumatismo por Reperfusão/imunologia , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Obstrução Ureteral/imunologia , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia
4.
J Pathol ; 241(1): 80-90, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27763657

RESUMO

Renal fibrosis is a significant threat to public health globally. Diverse primary aetiologies eventually result in chronic kidney disease (CKD) and immune cells influence this process. The roles of monocytes/macrophages, T cells, and mast cells have been carefully examined, whilst only a few studies have focused on the effect of B cells. We investigated B-cell function in tubulointerstitial fibrosis induced by unilateral ureteral obstruction (UUO), using genetic B-cell-deficient µMT mice or CD20 antibody-mediated B-cell-depleted mice. Obstructed kidneys of µMT and anti-CD20-treated mice showed lower levels of monocyte/macrophage infiltration and collagen deposition compared to wild-type mice. Mechanistically, anti-CD20 attenuated UUO-induced alterations of renal tumour necrosis factor-α (TNF-α), vascular cell adhesion molecule 1 (VCAM-1) pro-inflammatory genes, and CC chemokine ligand-2 (CCL2) essential for monocyte recruitment; B cells were one of the main sources of CCL2 in post-UUO kidneys. Neutralization of CCL2 reduced monocyte/macrophage influx and fibrotic changes in obstructed kidneys. Therefore, early-stage accumulation of B cells in the kidney accelerated monocyte/macrophage mobilization and infiltration, aggravating the fibrosis resulting from acutely induced kidney nephropathy. © 2016 The Authors. Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.


Assuntos
Linfócitos B/imunologia , Nefropatias/imunologia , Monócitos/imunologia , Obstrução Ureteral/imunologia , Animais , Movimento Celular/imunologia , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/imunologia , Quimiotaxia de Leucócito/imunologia , Fibrose , Mediadores da Inflamação/metabolismo , Nefropatias/etiologia , Túbulos Renais/imunologia , Túbulos Renais/patologia , Masculino , Camundongos Endogâmicos C57BL , Obstrução Ureteral/complicações
5.
Nephrology (Carlton) ; 23(6): 573-584, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28437591

RESUMO

AIM: We explored whether Fluorofenidone reduced interleukin-1ß (IL-1ß) production by interacting with NLRP3 inflammasome in unilateral ureteral obstruction (UUO). METHODS: Ureteral obstruction rats were treated with Fluorofenidone (500 mg/kg per day) for 3, 7 days. Morphologic analysis and leukocytes infiltration were assessed in ligated kidneys. Furthermore, plasmids of NLRP3, ASC, pro-Caspase-1, pro-IL-1ß were co-transfected into 293 T cells, and then treated with Fluorofenidone (2 mM). The expression of NLRP3, ASC, pro-caspase-1, cleavage caspase-1, pro-IL-1ß and cleavage IL-1ß were measured by Western blot or real-time PCR in vivo and in vitro. Moreover the interaction of NLRP3 inflammasome-assembly was detected by co-immunoprecipitation and confocal immunofluorescence. RESULTS: Fluorofenidone treatment significantly attenuated renal fibrosis and leukocytes infiltration in UUO model. Fluorofenidone had no effect on the expression of pro-IL-1ß. Interestingly, Fluorofenidone inhibited the activation of NLRP3 inflammasome, downregulated Caspase-1 levels and thereby decreased the cleavage of pro-IL-1ß into IL-1ß in vivo and in vitro. Fluorofenidone treatment distinctively weakened the interaction between NLRP3 and ASC, as well as ASC and pro-Caspase-1 in vivo. However, Fluorofenidone treatment only significantly weakened the interaction between ASC and pro-Caspase-1 in co-transfected 293 T cells. CONCLUSION: Fluorofenidone serves as a novel anti-inflammatory agent that attenuates IL-1ß production in UUO model by interacting with NLRP3 inflammasome.


Assuntos
Anti-Inflamatórios/farmacologia , Inflamassomos/efeitos dos fármacos , Interleucina-1beta/metabolismo , Rim/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nefrite/prevenção & controle , Piridonas/farmacologia , Obstrução Ureteral/tratamento farmacológico , Animais , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Caspase 1/metabolismo , Quimiotaxia de Leucócito/efeitos dos fármacos , Modelos Animais de Doenças , Regulação para Baixo , Fibrose , Células HEK293 , Humanos , Inflamassomos/genética , Inflamassomos/imunologia , Inflamassomos/metabolismo , Interleucina-1beta/genética , Rim/imunologia , Rim/metabolismo , Rim/patologia , Masculino , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Nefrite/imunologia , Nefrite/metabolismo , Nefrite/patologia , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos , Fatores de Tempo , Obstrução Ureteral/imunologia , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia
6.
Clin Exp Immunol ; 190(1): 68-78, 2017 10.
Artigo em Inglês | MEDLINE | ID: mdl-28658504

RESUMO

Renal fibrosis is the common final manifestation of chronic kidney diseases and usually results in end-stage renal failure. In this study, we evaluated the effect of fingolimod (FTY720), an analogue of sphingosine 1-phosphate (S1P), as a treatment for the unilateral ureteral obstruction (UUO)-induced renal fibrosis animal model. We treated mice with FTY720 at a dosage of 1 mg/kg/day by intragastric administration from day 1 until day 7. The control group received the same amount of saline. FTY720 reduced significantly the urine albumin/creatinine ratio (UACR) in treated UUO mice. FTY720 treatment also caused a significant decrease in interstitial expansion and collagen deposition in the kidney, accompanied by reduced mononuclear cell recruitment and inflammatory cytokine expression. In addition, the expression levels of the endothelial cell adhesion molecules P-selectin and vascular cell adhesion protein 1 (VCAM-1) were suppressed in the ligated kidney by FTY720 administration, suggesting reduced renal endothelial cell activation. Furthermore, in renal interstitial fibroblast normal rat kidney (NRK)-49F cells, FTY720 significantly affected transforming growth factor (TGF)-ß-induced α-smooth muscle actin (SMA) expression and collagen synthesis by inhibiting both the Mothers against decapentaplegic homologue (Smad)2/3 and phosphatidylinositol 3-kinase/protein kinase B/glycogen synthase kinase 3 beta (PI3K/AKT/GSK3ß) signalling pathways. S1P1 knock-down by siRNA reversed this effect significantly in our fibroblast cell culture model. Therefore, FTY720 attenuates renal fibrosis via two different mechanisms: first, FTY720 suppresses the synthesis of extracellular matrix in interstitial fibroblasts by interfering with TGF-ß signalling; and secondly, FTY720 affects endothelial cell activation and chemokine expression, thereby reducing immune cell recruitment into the kidney.


Assuntos
Fibroblastos/efeitos dos fármacos , Cloridrato de Fingolimode/uso terapêutico , Imunossupressores/uso terapêutico , Rim/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo , Obstrução Ureteral/tratamento farmacológico , Animais , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Creatinina/urina , Matriz Extracelular/metabolismo , Fibroblastos/imunologia , Fibrose , Humanos , Rim/metabolismo , Rim/patologia , Leucócitos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , RNA Interferente Pequeno/genética , Ratos , Receptores de Lisoesfingolipídeo/genética , Receptores de Lisoesfingolipídeo/metabolismo , Transdução de Sinais/efeitos dos fármacos , Obstrução Ureteral/imunologia
7.
J Immunol ; 195(6): 2797-805, 2015 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-26254342

RESUMO

Chemokines modulate inflammatory responses that are prerequisites for kidney injury. The specific role of monocyte-associated CX3CR1 and its cognate ligand CX3CL1 in unilateral ureteral obstruction (UUO)-induced kidney injury remains unclear. In this study, we found that UUO caused a CCR2-dependent increase in numbers of Ly6C(hi) monocytes both in the blood and kidneys and of Ly6C(-)CX3CR1(+) macrophages in the obstructed kidneys of mice. Using CX3CR1(gfp/+) knockin mice, we observed a rapid conversion of infiltrating proinflammatory Ly6C(+)CX3CR1(1o) monocytes/macrophages to anti-inflammatory Ly6C(-)CX3CR1(hi) macrophages. CX3CR1 deficiency affected neither monocyte trafficking nor macrophage differentiation in vivo upon renal obstruction, but CX3CR1 expression in monocytes and macrophages was required for increases in fibrosis in the obstructed kidneys. Mechanistically, CX3CL1-CX3CR1 interaction increases Ly6C(-)CX3CR1(hi) macrophage survival within the obstructed kidneys. Therefore, CX3CL1 and CX3CR1 may represent attractive therapeutic targets in obstructive nephropathy.


Assuntos
Quimiocina CX3CL1/metabolismo , Fibrose/patologia , Macrófagos/imunologia , Receptores de Quimiocinas/metabolismo , Obstrução Ureteral/patologia , Animais , Antígenos Ly/metabolismo , Receptor 1 de Quimiocina CX3C , Diferenciação Celular/imunologia , Movimento Celular/imunologia , Células Cultivadas , Fibrose/imunologia , Rim/lesões , Rim/patologia , Nefropatias/imunologia , Macrófagos/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Receptores CCR2/metabolismo , Receptores de Quimiocinas/genética , Transdução de Sinais/imunologia , Obstrução Ureteral/imunologia
8.
Kidney Int ; 89(4): 809-22, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26994575

RESUMO

Scavenger receptor CD36 participates in lipid metabolism and inflammatory pathways important for cardiovascular disease and chronic kidney disease (CKD). Few pharmacological agents are available to slow the progression of CKD. However, apolipoprotein A-I-mimetic peptide 5A antagonizes CD36 in vitro. To test the efficacy of 5A, and to test the role of CD36 during CKD, we compared wild-type to CD36 knockout mice and wild-type mice treated with 5A, in a progressive CKD model that resembles human disease. Knockout and 5A-treated wild-type mice were protected from CKD progression without changes in blood pressure and had reductions in cardiovascular risk surrogate markers that are associated with CKD. Treatment with 5A did not further protect CD36 knockout mice from CKD progression, implicating CD36 as its main site of action. In a separate model of kidney fibrosis, 5A-treated wild-type mice had less macrophage infiltration and interstitial fibrosis. Peptide 5A exerted anti-inflammatory effects in the kidney and decreased renal expression of inflammasome genes. Thus, CD36 is a new therapeutic target for CKD and its associated cardiovascular risk factors. Peptide 5A may be a promising new agent to slow CKD progression.


Assuntos
Antígenos CD36/antagonistas & inibidores , Peptídeos/uso terapêutico , Insuficiência Renal Crônica/prevenção & controle , Angiotensina II , Animais , Pressão Sanguínea , Quimiocina CXCL1/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Avaliação Pré-Clínica de Medicamentos , Fibrose , Corantes Fluorescentes , Células HeLa , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Rim/imunologia , Rim/patologia , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Nefrectomia , Peptídeos/farmacologia , Insuficiência Renal Crônica/metabolismo , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
9.
Cell Physiol Biochem ; 39(5): 1837-1849, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27744426

RESUMO

AIMS: Sorafenib, which has been used extensively for the treatment of renal cell cancer and advanced hepatocellular carcinoma (HCC), has also been shown to have antifibrotic effects in liver fibrosis. However, the effects of sorafenib on renal fibrosis are unknown. Therefore, we investigated whether sorafenib inhibited renal fibrosis in a mouse model of unilateral ureteral obstruction (UUO) and further explored the potential mechanism. METHODS: Mice underwent UUO followed by vehicle or sorafenib treatment. The expression of CD68, a macrophage marker, and the pro-inflammatory cytokines, MCP1 and CXCR3, were immunohistochemically analyzed. The involvement of macrophages in the formation of renal fibrosis was studied using confocal microscopy. RESULTS: Renal histopathology improved in the UUO-sorafenib mice. Sorafenib notably suppressed TGF-ß1-mediated renal fibrogenic effects. The mRNA and protein expressions of CD68, MCP1, and CXCR3 in the obstructed kidney were significantly decreased by sorafenib. Immunohistochemistry showed that CD68 and CXCR3 had a similar distribution, whereas MCP1 was observed predominantly in the tubular epithelial cells. Double immunofluorescence demonstrated that CD68-positive macrophages could co-localize with CXCR3. It also revealed that CXCR3 interacted with CXCL11 in the UUO mouse kidneys. Widespread adhesion of macrophages to myofibroblasts was markedly inhibited in UUO-sorafenib mouse kidneys. CONCLUSIONS: Taken together, the results indicated that sorafenib had protective effects against renal fibrosis; its mechanism of action was associated with inhibition of macrophage infiltration via the CXCR3/CXCL11 pathway. These data suggest the clinical potential of sorafenib for treatment of renal fibrosis and illustrate the immunological mechanisms underlying the protective effects of sorafenib.


Assuntos
Antineoplásicos/farmacologia , Movimento Celular/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Niacinamida/análogos & derivados , Compostos de Fenilureia/farmacologia , Obstrução Ureteral/tratamento farmacológico , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/genética , Antígenos de Diferenciação Mielomonocítica/imunologia , Adesão Celular/efeitos dos fármacos , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Quimiocina CXCL11/genética , Quimiocina CXCL11/imunologia , Modelos Animais de Doenças , Fibrose/prevenção & controle , Regulação da Expressão Gênica , Humanos , Túbulos Renais/efeitos dos fármacos , Túbulos Renais/imunologia , Túbulos Renais/patologia , Macrófagos/imunologia , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miofibroblastos/efeitos dos fármacos , Miofibroblastos/imunologia , Miofibroblastos/patologia , Niacinamida/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Receptores CXCR3/antagonistas & inibidores , Receptores CXCR3/genética , Receptores CXCR3/imunologia , Transdução de Sinais , Sorafenibe , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/imunologia , Ureter/efeitos dos fármacos , Ureter/imunologia , Ureter/patologia , Obstrução Ureteral/genética , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
10.
Am J Physiol Renal Physiol ; 308(1): F69-75, 2015 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-25377911

RESUMO

Previous studies have indicated that macrophage phenotype diversity is involved in the progression of renal fibrosis. However, the factors facilitating M1 or M2 phenotypes and the function of these polarized macrophages in kidney injury and fibrosis remain largely unknown. In the present study, we found that macrophages accumulated in the kidney interstitium exhibited mainly as the M1 phenotype at the early stage of unilateral ureter obstruction (UUO). High-mobility group box 1 (HMGB1) protein expressed and released from tubular epithelial cells and interstitial macrophages was essential for the M1 macrophage transition. HMGB1 significantly induced the expression of the M1 marker inducible nitric oxide synthase while decreasing the M2 marker IL-10 in macrophages. Moreover, a glycyrrhizic acid derivative, a blocker of HMGB1 release, reduced UUO-mediated kidney injury and ameliorated UUO-induced renal fibrosis. Interestingly and importantly, UUO caused a low pH value in the urine accumulated in the obstructed ureter, and the acidified urine induced HMGB1 release from tubular epithelial cells and macrophages in vitro. Our data demonstrate that HMGB1 is an essential contributor in facilitating M1 polarization at the early stage of UUO. Inhibition of HMGB1 release may alter macrophage phenotype and contribute to the protection of kidney tissue from injury and fibrosis.


Assuntos
Proteína HMGB1/metabolismo , Macrófagos/fisiologia , Nefroesclerose/imunologia , Obstrução Ureteral/imunologia , Animais , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos , Ácido Glicirrízico/farmacologia , Ácido Glicirrízico/uso terapêutico , Proteína HMGB1/antagonistas & inibidores , Humanos , Masculino , Camundongos Endogâmicos C57BL , Nefroesclerose/tratamento farmacológico , Nefroesclerose/metabolismo , Fenótipo , Fitoterapia , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Obstrução Ureteral/complicações , Obstrução Ureteral/metabolismo
12.
J Tradit Chin Med ; 35(5): 564-70, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26591687

RESUMO

OBJECTIVE: To investigate the effect of salvianolic acid A and C component molecules, which are involved in drug compatibility, on inflammatory cytokine expression that affects human chemokine ligand 5 (CCL5) and chemokine ligand 10 (CXCL10) levels in rats with unilateral ureteral obstruction (UUO). METHODS: Fifty Sprague Dawley rats were randomly divided into five groups: normal, model, salvianolic acid A, salvianolic acid C and salvianolic acid A and C groups. The normal group was used as the control, and the other groups of rats had a UUO model established. The control group had free access to food and water, and the other groups received the corresponding drugs for 2 weeks. After the last administration, urine ß2-microglobulin (ß 2-MG) and N-acetyl-ß-D-glucosaminidase (NAG) levels were analyzed. After 24 h, all rats were sacrificed and the serum was analyzed for creatinine (Cr) and blood urea nitrogen (BUN) levels. Rat kidneys were removed, and CCL5 and CXCL10 inflammatory cytokine mRNA expression was measured using real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-PCR). Kidney fibrosis was observed using hematoxylin-eosin (HE) staining and Masson trichrome staining. RESULTS: In the salvianolic acid A and salvianolic acid C treatment groups, serum Cr and urine NAG levels were significantly lower than in the model group (both P < 0.05). In all treatment groups, urine ß2-MG levels were significantly lower than in the model group (all P < 0.05). Compared with model group, the pathological changes and collagen deposition improved to varying degrees (both P < 0.05). CCL5 and CXCL10 mRNA expression decreased to different degrees compared with the model group (both P < 0.05). CONCLUSION: Salvianolic acid A and C are component molecules of drug compatibility, and they may protect renal function and improve tubular function and renal pathology to a certain degree in UUO. This improvement may be related to a reduction in inflammatory cytokines CCL5 and CXCL10 secretion in the UUO rat kidney.


Assuntos
Alcenos/administração & dosagem , Ácidos Cafeicos/administração & dosagem , Citocinas/imunologia , Medicamentos de Ervas Chinesas/administração & dosagem , Lactatos/administração & dosagem , Polifenóis/administração & dosagem , Obstrução Ureteral/tratamento farmacológico , Animais , Quimiocina CCL5/genética , Quimiocina CCL5/imunologia , Quimiocina CXCL10/genética , Quimiocina CXCL10/imunologia , Citocinas/genética , Humanos , Masculino , Ratos , Obstrução Ureteral/genética , Obstrução Ureteral/imunologia
13.
J Immunol ; 188(10): 5106-15, 2012 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-22490864

RESUMO

Renal fibrosis and inflammation are associated with hypoxia, and tissue pO(2) plays a central role in modulating the progression of chronic kidney disease. Key mediators of cellular adaptation to hypoxia are hypoxia-inducible factor (HIF)-1 and -2. In the kidney, they are expressed in a cell type-specific manner; to what degree activation of each homolog modulates renal fibrogenesis and inflammation has not been established. To address this issue, we used Cre-loxP recombination to activate or to delete both Hif-1 and Hif-2 either globally or cell type specifically in myeloid cells. Global activation of Hif suppressed inflammation and fibrogenesis in mice subjected to unilateral ureteral obstruction, whereas activation of Hif in myeloid cells suppressed inflammation only. Suppression of inflammatory cell infiltration was associated with downregulation of CC chemokine receptors in renal macrophages. Conversely, global deletion or myeloid-specific inactivation of Hif promoted inflammation. Furthermore, prolonged hypoxia suppressed the expression of multiple inflammatory molecules in noninjured kidneys. Collectively, we provide experimental evidence that hypoxia and/or myeloid cell-specific HIF activation attenuates renal inflammation associated with chronic kidney injury.


Assuntos
Injúria Renal Aguda/imunologia , Injúria Renal Aguda/patologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/fisiologia , Células Mieloides/imunologia , Células Mieloides/patologia , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia , Injúria Renal Aguda/genética , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/deficiência , Modelos Animais de Doenças , Fibrose/imunologia , Fibrose/prevenção & controle , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/deficiência , Inflamação/imunologia , Inflamação/patologia , Inflamação/prevenção & controle , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Células Mieloides/metabolismo , Cultura Primária de Células , Obstrução Ureteral/genética
14.
Am J Physiol Renal Physiol ; 305(10): F1445-54, 2013 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-24026183

RESUMO

CD44 family members are cell surface glycoproteins, which are expressed on tubular epithelial cells (TEC) solely upon kidney injury and are involved in renal fibrosis development. Renal interstitial fibrosis is the final manifestation of chronic kidney diseases and is regulated by a complex network of cytokines, including the profibrotic factor transforming growth factor-ß1 (TGF-ß1) and the two antifibrotic cytokines bone morphogenic protein-7 (BMP-7) and hepatocyte growth factor (HGF). The present study investigates the potential role of CD44 standard (CD44s) and CD44v3-v10 (CD44v3) isoforms as modulators of the balance between TGF-ß1 and HGF/BMP-7. CD44s is the shortest and most common isoform. CD44v3-v10 (CD44v3) has heparan sulfate moieties, which enable the binding to HGF/BMP-7, and hence, might exert renoprotective effects. Using transgenic mice overexpressing either CD44s or CD44v3 specifically on proximal TEC, we found that in vitro the overexpression of CD44v3 on primary TEC renders cells less susceptible to TGF-ß1 profibrotic actions and more sensitive to BMP-7 and HGF compared with TEC overexpressing CD44s. One day after unilateral ureteric obstruction, obstructed kidneys from CD44v3 transgenic mice showed less tubular damage and myofibroblasts accumulation, which was associated with decreased TGF-ß1 signaling and increased BMP-7 synthesis and signaling compared with kidneys from wild-type and CD44s transgenic mice. These data suggest that CD44v3 plays a renoprotective role in early stage of chronic obstructive nephropathy.


Assuntos
Receptores de Hialuronatos/metabolismo , Túbulos Renais Proximais/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Obstrução Ureteral/prevenção & controle , Animais , Proteína Morfogenética Óssea 7/metabolismo , Células Cultivadas , Doença Crônica , Modelos Animais de Doenças , Fibrose , Fator de Crescimento de Hepatócito/metabolismo , Receptores de Hialuronatos/genética , Mediadores da Inflamação/metabolismo , Túbulos Renais Proximais/imunologia , Túbulos Renais Proximais/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Miofibroblastos/metabolismo , Proteínas Recombinantes/metabolismo , Transdução de Sinais , Fatores de Tempo , Regulação para Cima , Obstrução Ureteral/genética , Obstrução Ureteral/imunologia , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia
15.
Kidney Int ; 84(2): 317-26, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23515052

RESUMO

Mast cell release of chymase is important in tissue remodeling and may participate in inflammation leading to fibrosis and organ failure. Here we analyzed the function of chymase in unilateral ureteral obstruction, an established accelerated model of renal tubulointerstitial fibrosis. Mice deficient in mouse mast cell protease 4 (mMCP4), the functional counterpart of human chymase, had increased obstruction-induced fibrosis when compared to wild-type mice indicating a protective effect of mMCP4. Engraftment of mast cell-deficient Kit(Wsh/Wsh) mice with wild type, but not mMCP4-deficient mast cells, restored protection confirming the role of mMCP4. Kidneys of mMCP4-deficient mice had higher levels of renal tubular damage, interstitial fibrosis, collagen deposition, increased α-smooth muscle actin, and decreased E-cadherin expression compared to the kidneys of wild-type mice. Further analysis showed an elevated inflammatory response in mMCP4-deficient mice with increased levels of kidney-infiltrating macrophages and T cells and local profibrotic TGF-ß1 and CCL2. Granulated and degranulated mast cells and mMCP4 were mainly found in the kidney capsule, respectively, before and after ureteral obstruction. Analysis of mMCP4 substrates showed that it mediates its anti-fibrotic actions by degrading interstitial deposits of fibronectin, a known promoter of inflammatory cell infiltration and adhesion. Thus, mast cell released mMCP4 has anti-fibrotic potential in acutely induced obstructive nephropathy.


Assuntos
Quimases/metabolismo , Nefropatias/prevenção & controle , Rim/enzimologia , Mastócitos/enzimologia , Serina Endopeptidases/metabolismo , Obstrução Ureteral/complicações , Actinas/metabolismo , Animais , Caderinas/metabolismo , Degranulação Celular , Quimiocina CCL2/metabolismo , Quimiotaxia , Quimases/deficiência , Quimases/genética , Colágeno/metabolismo , Modelos Animais de Doenças , Fibronectinas/metabolismo , Fibrose , Rim/imunologia , Rim/patologia , Nefropatias/enzimologia , Nefropatias/etiologia , Nefropatias/imunologia , Nefropatias/patologia , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Mastócitos/imunologia , Mastócitos/transplante , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Miofibroblastos/metabolismo , Serina Endopeptidases/deficiência , Serina Endopeptidases/genética , Linfócitos T/imunologia , Linfócitos T/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Obstrução Ureteral/enzimologia , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
16.
Am J Pathol ; 180(1): 91-103, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22079432

RESUMO

Unilateral ureteral obstruction (UUO) is a well-characterized murine model of renal inflammation leading to fibrosis. Renal dendritic cells (DCs) constitute a significant portion of kidney leukocytes and may participate in local inflammation and have critical roles in antigen presentation. The heterogeneity in renal DC populations and surface marker overlap with monocytes/macrophages has made studying renal DCs difficult. These studies used CD11c-promoter driven reporter/depletion mice to study DCs in vivo. Studying early local inflammatory events (day 3 of UUO), in vivo multiphoton imaging of the intact kidney of CD11c reporter mice revealed more dendrite extensions and increased activity of renal DCs in real time. Phenotypic analysis suggested resident DC maturation in obstructed kidneys with increased CD11b and less F4/80 expressed. CD11b(hi) Gr-1(+) inflammatory DCs were also present in obstructed kidneys. T-cell receptor transgenic mice revealed enhanced antigen-presenting capacity of renal DCs after UUO, with increased antigen-specific T-cell proliferation in vivo and ex vivo. However, conditional DC ablation at days 0, 2, or 4 did not attenuate fibrosis or apoptosis 7 days after UUO, and depletion at 7 days did not alter outcomes at day 14. Therefore, after UUO, renal DCs exhibit inflammatory morphological and functional characteristics and are more effective antigen-presenting cells, but they do not directly contribute to tubulointerstitial damage and fibrosis.


Assuntos
Células Dendríticas/fisiologia , Ativação Linfocitária/imunologia , Linfócitos T/imunologia , Ureter/patologia , Obstrução Ureteral/patologia , Animais , Apoptose/imunologia , Antígeno CD11c/metabolismo , Citocinas/biossíntese , Células Dendríticas/metabolismo , Fibrose , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Nefrite/patologia , Fenótipo , Obstrução Ureteral/imunologia
17.
Zhongguo Dang Dai Er Ke Za Zhi ; 15(12): 1134-8, 2013 Dec.
Artigo em Zh | MEDLINE | ID: mdl-24342215

RESUMO

OBJECTIVE: To study the expression of Fractalkine (FKN) in the kidney tissue of rats with renal fibrosis and the effect of IL-18BP on FKN. METHODS: Male Wister rats were randomly assigned to sham-operation (n=24), unilatral ureteral obstruction (UUO, n=22), and IL-18 binding protein (IL-18BP) treatment groups (n=23). The UUO model was prepared by unilateral ureteral ligation in the later two groups. The IL-18BP treatment group received an intraperitoneal injection of IL-18BP (0.1 mg/kg) every other day after UUO inducement, for 7 times, while normal saline was administered in the other two groups. Seven or eight rats of every group were sacrificed at 3, 7 or 14 days after IL-18BP or normal saline injections. FKN levels at various times were detected by immunohistochemistry and Western blot. RESULTS: Compared with the sham-operation group, FKN levels in the kidney tissue of the untreated UUO group increased significantly at all time points (P<0.01). IL-18BP treatment decreased significantly FKN levels in the kidney tissue at all time points compared with the untreated UUO group (P<0.01). CONCLUSIONS: IL-18BP treatment may down-regulate the increased FKN levels of the rat kidney tissue caused by UUO, possibly thus delays the occurrence and development of renal fibrosis.


Assuntos
Quimiocina CX3CL1/genética , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Rim/patologia , Animais , Western Blotting , Fibrose , Rim/imunologia , Masculino , Ratos , Ratos Wistar , Obstrução Ureteral/imunologia
18.
Kidney Int ; 81(4): 379-90, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21975862

RESUMO

Interleukin 17A-secreting T-helper 17 (Th17) cells are pathogenic in inflammatory kidney diseases, but their intrarenal regulation is poorly understood. In order to better define Th17 cell dynamics during interstitial inflammation, we utilized the mouse unilateral ureteral obstruction model to analyze inflammatory cell subtypes by multicolor flow cytometry and cell sorting and by effects on in vitro-generated Th17 cells. Interleukin 17A expression localized to CCR6(+)CCR4(+/-)CD4(+) T-cells and progressively increased in obstructed kidneys. The number of CCR6(+)CD4(+) T-cells increased over 10-fold by 72 h, were enriched for interleukin 17A production, and were highly proliferative based on in vivo bromodeoxyuridine incorporation. Secreted products of leukocytes isolated from obstructed kidneys enhanced the interleukin 17A production of in vitro-generated Th17 cells. This Th17-enhancing activity was identified as interleukin-1 produced by renal dendritic cells and monocytes. The in vivo validity of these findings was confirmed in mice lacking the interleulin-1 receptor and in mice treated with a recombinant interleukin-1 receptor antagonist, each of which exhibited reduced intrarenal Th17 activity compared with control mice. Thus, the inflamed kidney accumulates CCR6(+) Th17 cells that undergo activation and proliferation. Production of interleukin 1 family cytokines by resident dendritic cells and infiltrating monocytes enhances intrarenal Th17 activation in acute kidney injury.


Assuntos
Interleucina-17/metabolismo , Interleucina-1/imunologia , Nefrite/imunologia , Células Th17/imunologia , Obstrução Ureteral/imunologia , Animais , Antígenos CD4/análise , Proliferação de Células , Células Cultivadas , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Feminino , Citometria de Fluxo , Interleucina-1/metabolismo , Contagem de Linfócitos , Camundongos , Camundongos Endogâmicos C57BL , Monócitos/metabolismo , Nefrite/metabolismo , Receptores CCR4/análise , Receptores CCR6/análise , Obstrução Ureteral/complicações , Obstrução Ureteral/metabolismo
19.
Kidney Int ; 82(6): 676-85, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22673890

RESUMO

Progressive renal fibrosis is the final common pathway leading to renal failure irrespective of the initiating cause. Clinical studies of renal fibrosis found that prominent mast cell accumulation correlated with worse outcomes. Mast cells are pluripotent innate immune cells that synthesize and secrete profibrotic mediators. Here we use mast cell-deficient (Kit(W-sh/W-sh)) mice to define a functional pathogenic role for these cells in the development of renal fibrosis. Intrarenal collagen deposition was significantly decreased in mast cell-deficient compared to wild-type mice 7 and 14 days after unilateral ureteric obstruction. The intrarenal expression of mRNAs for transforming growth factor-ß, α-smooth muscle actin, chemokines, and renal macrophages and CD4(+) T cells were also decreased in mast cell-deficient mice. Reconstitution of the mast cell population in mast cell-deficient mice with wild-type bone marrow-derived mast cells restored the pattern and intensity of renal fibrosis to levels seen in wild-type mice following ureteric ligation. Interestingly, the mast cells were recruited, activated, and degranulated within 6 h of ureteric ligation. A mast cell stabilizer that impairs degranulation, disodium chromoglycate, significantly attenuated renal fibrosis following ureteric ligation in wild-type mice. Thus, mast cells promote renal fibrosis and their targeting may offer therapeutic potential in the treatment of renal fibrosis.


Assuntos
Degranulação Celular , Nefropatias/etiologia , Rim/imunologia , Mastócitos/imunologia , Obstrução Ureteral/complicações , Actinas/genética , Actinas/metabolismo , Animais , Linfócitos T CD4-Positivos/imunologia , Degranulação Celular/efeitos dos fármacos , Células Cultivadas , Quimiocinas/genética , Quimiocinas/metabolismo , Colágeno/metabolismo , Cromolina Sódica/farmacologia , Modelos Animais de Doenças , Progressão da Doença , Fibrose , Regulação da Expressão Gênica , Hidronefrose/etiologia , Hidronefrose/imunologia , Hidronefrose/patologia , Rim/efeitos dos fármacos , Rim/patologia , Nefropatias/genética , Nefropatias/imunologia , Nefropatias/patologia , Nefropatias/prevenção & controle , Macrófagos/imunologia , Masculino , Mastócitos/efeitos dos fármacos , Mastócitos/transplante , Metaloproteinase 12 da Matriz/genética , Metaloproteinase 12 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mutação , Fenótipo , Proteínas Proto-Oncogênicas c-kit/genética , Fatores de Tempo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Obstrução Ureteral/genética , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
20.
Mol Med ; 18: 1231-9, 2012 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-22777483

RESUMO

Inflammation contributes to the pathogenesis of chronic kidney disease (CKD). Molecules released by the inflamed injured tissue can activate toll-like receptors (TLRs), thereby modulating macrophage and CD4(+) T-cell activity. We propose that in renal fibrogenesis, M2 macrophages are recruited and activated in a T helper subset 2 cell (T(H)2)-prone inflammatory milieu in a MyD88-dependent manner. Mice submitted to unilateral ureteral ligation (UUO) demonstrated an increase in macrophage infiltration with collagen deposition after 7 d. Conversely, TLR2, TLR4 and MyD88 knockout (KO) mice had an improved renal function together with diminished T(H)2 cytokine production and decreased fibrosis formation. Moreover, TLR2, TLR4 and MyD88 KO animals exhibited less M2 macrophage infiltration, namely interleukin (IL)-10(+) and CD206(+) CD11b(high) cells, at 7 d after surgery. We evaluated the role of a T(H)2 cytokine in this context, and observed that the absence of IL-4 was associated with better renal function, decreased IL-13 and TGF-ß levels, reduced arginase activity and a decrease in fibrosis formation when compared with IL-12 KO and wild-type (WT) animals. Indeed, the better renal outcomes and the decreased fibrosis formation were restricted to the deficiency of IL-4 in the hematopoietic compartment. Finally, macrophage depletion, rather than the absence of T cells, led to reduced lesions of the glomerular filtration barrier and decreased collagen deposition. These results provide evidence that future therapeutic strategies against renal fibrosis should be accompanied by the modulation of the M1:M2 and T(H)1:T(H)2 balance, as T(H)2 and M2 cells are predictive of fibrosis toward mechanisms that are sensed by innate immune response and triggered in a MyD88-dependent pathway.


Assuntos
Imunidade/imunologia , Rim/patologia , Ativação de Macrófagos/imunologia , Macrófagos/imunologia , Fator 88 de Diferenciação Mieloide/metabolismo , Transdução de Sinais/imunologia , Células Th2/imunologia , Animais , Citocinas/metabolismo , Fibrose , Hematopoese , Interleucina-12/metabolismo , Interleucina-4/deficiência , Rim/imunologia , Rim/fisiopatologia , Nefropatias/imunologia , Nefropatias/patologia , Nefropatias/fisiopatologia , Testes de Função Renal , Ligadura , Macrófagos/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptor 2 Toll-Like/metabolismo , Receptor 4 Toll-Like/metabolismo , Ureter/patologia , Obstrução Ureteral/complicações , Obstrução Ureteral/imunologia , Obstrução Ureteral/patologia
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