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1.
Genes Cells ; 28(7): 482-495, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37073980

RESUMO

Histone methylation plays a vital role in retinal development. However, the role of histone H3K36 methylation in retinal development is not clear. We examined the role of H3K36 methylation by loss-of-function analysis of H3K36me1/2 demethylases, Fbxl10, and Fbxl11. We analyzed the effect of knockout of these genes in the developing and mature retina on retinal development. Knockout of Fbxl10 specifically in the developing retina did not result in gross developmental abnormalities. Although adult rod photoreceptor-specific knockout of Fbxl11 in mature retinas did not result in morphological abnormalities, Fbxl11 knockout in developing retinas increased apoptosis, suppressed the proliferation of retinal progenitor cells, and resulted in microphthalmia. Morphological analysis revealed perturbed differentiation of rod photoreceptor and bipolar cells. RNA-seq of retinas at P7 showed markedly decreased expression of genes characterizing rod photoreceptor and bipolar cells in Fbxl11-knockout retinas. In addition, perturbation of alternative splicing increased intron retention in Fbxl11-knockout retinas. Genome-wide evaluation of the H3K36 methylation status revealed that Fbxl11 knockout altered the distribution of H3K36me2/3 in genes important for rod photoreceptor development. Taken together, we show that Fbxl11 plays pivotal roles in the development of retinal late-born cell types and may contribute to tight control of H3K36 methylation during retinal development.


Assuntos
Histona Desmetilases , Histonas , Diferenciação Celular/genética , Histona Desmetilases/genética , Histonas/genética , Histonas/metabolismo , Retina/metabolismo , Células Fotorreceptoras Retinianas Bastonetes , Animais , Camundongos
2.
Nephrology (Carlton) ; 16(1): 76-86, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21175982

RESUMO

AIM: Transforming growth factor-ß (TGF-ß) is involved in renal tubulointerstitial fibrosis. Recently, the ubiquitin proteasome system was shown to participate in the TGF-ß signalling pathway. The aim of this study was to examine the effects of proteasome inhibitors on TGF-ß-induced transformation of renal fibroblasts and tubular epithelial cells in vitro and on unilateral ureteral obstruction (UUO) in vivo. METHODS: Rat renal fibroblasts NRK-49F cells and tubular epithelial cells, NRK-52E, were treated with TGF-ß in the presence or absence of a proteasome inhibitor, MG132 or lactacystin. Rats were subjected to UUO and received MG132 i.p. for 7 days. RESULTS: In cultured renal cells, both MG132 and lactacystin inhibited TGF-ß-induced α-smooth muscle actin (α-SMA) protein expression according to both western blotting and immunofluorescent study results. MG132 also suppressed TGF-ß-induced mRNA expression of α-SMA and upregulation of Smad-response element reporter activity. However, MG132 did not inhibit TGF-ß-induced phosphorylation and nuclear translocation of Smad2. In contrast, MG132 increased the protein level of Smad co-repressor SnoN, demonstrating that SnoN is one of the target molecules by which MG132 blocks the TGF-ß signalling pathway. Although the proteasome inhibitor suppressed TGF-ß-induced transformation of cultured fibroblasts and tubular epithelial cells, MG132 treatment did not ameliorate tubulointerstitial fibrosis in the rat UUO model. CONCLUSION: Proteasome inhibitors attenuate TGF-ß signalling by blocking Smad signal transduction in vitro, but do not inhibit renal interstitial fibrosis in vivo.


Assuntos
Inibidores de Cisteína Proteinase/farmacologia , Fibrose/metabolismo , Nefropatias/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Fator de Crescimento Transformador beta/metabolismo , Acetilcisteína/análogos & derivados , Acetilcisteína/farmacologia , Actinas/genética , Actinas/metabolismo , Animais , Células Cultivadas , Modelos Animais de Doenças , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Fibroblastos/citologia , Fibroblastos/metabolismo , Fibrose/genética , Nefropatias/patologia , Leupeptinas/farmacologia , Masculino , Proteínas do Tecido Nervoso/metabolismo , Ratos , Ratos Wistar , Transdução de Sinais/genética , Proteínas Smad Reguladas por Receptor/metabolismo , Proteína Smad2/metabolismo , Proteína Smad3/metabolismo , Proteína Smad4/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica/efeitos dos fármacos , Fator de Crescimento Transformador beta/efeitos dos fármacos , Obstrução Ureteral/metabolismo , Obstrução Ureteral/patologia
3.
Nephron Exp Nephrol ; 116(1): e3-10, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20502051

RESUMO

BACKGROUND/AIMS: Mizoribine (MZR) is an imidazole nucleoside used as a therapeutic immunosuppressive agent. Though a previous report showed that MZR ameliorates proteinuria in puromycin aminonucleoside (PAN) nephropathy, the effect of MZR on podocytes has not been clarified. In this study, we determined whether MZR directly prevents PAN-induced podocyte injury. METHODS: Rats were intravenously injected once on day 0 with 100 mg/kg of PAN and received daily subcutaneous injections of MZR at a dose of 10 mg/kg from days 0 to 14. Cultured podocytes were pretreated with 50 microg/ml of MZR and then treated with 30 microg/ml of PAN. RESULTS: In rat PAN nephrosis, treatment with MZR from days 0 to 14 almost completely inhibited proteinuria. Immunofluorescence staining of nephrin was diminished, showing a discontinuous pattern in saline-treated PAN rats. In contrast, MZR treatment resulted in maintenance of a normal linear pattern. In cultured podocytes exposed to PAN, the percentages of viable cells were significantly increased with MZR treatment. The protective effect of MZR on PAN-induced podocyte injury was independent of inosine 5'-monophosphate dehydrogenase that is a known target enzyme of MZR as an immunosuppressant. MZR reduced PAN-induced integrin-linked kinase activation (ILK) and phosphorylation of glycogen synthase kinase-3beta (GSK3beta) in vivo and in vitro. CONCLUSION: MZR directly prevents PAN-induced podocyte injury, possibly by affecting signaling cascades involving ILK and GSK3beta.


Assuntos
Imunossupressores/uso terapêutico , Nefrose/tratamento farmacológico , Podócitos/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Ribonucleosídeos/uso terapêutico , Animais , IMP Desidrogenase/antagonistas & inibidores , IMP Desidrogenase/metabolismo , Masculino , Proteínas de Membrana/biossíntese , Nefrose/induzido quimicamente , Puromicina Aminonucleosídeo/antagonistas & inibidores , Ratos , Ratos Wistar
4.
Nephrology (Carlton) ; 15(5): 568-74, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20649878

RESUMO

AIM: Nestin, an intermediate filament originally identified as a marker of neural progenitor cells, is transiently expressed in endothelial cells and tubuloepithelial cells during kidney development. However, in adult kidneys, podocytes are the only cells that express nestin. In this study, we examined tubulointerstitial nestin expression in human glomerulonephritis. METHODS: Renal biopsy specimens obtained from 41 adult patients with immunoglobulin (Ig)A nephropathy were studied. Nestin expression was determined by immunohistochemical staining and estimated by digital image analysis. To identify the phenotype of nestin-positive cells, a double immunofluorescent study was performed for nestin and CD34 (a marker for endothelial cells) or alpha-smooth muscle actin (alpha-SMA, a marker for myofibroblasts). RESULTS: In normal kidney, nestin expression was restricted to the podocytes and was not detected in tubular cells and tubulointerstitial cells. In contrast, increased nestin expression was observed at tubulointerstitial areas of IgA nephropathy. The degree of tubulointerstitial nestin expression was positively correlated with tubulointerstitial fibrosis (r = 0.546, P < 0.001). The double immunofluorescent study showed that most nestin-positive cells in the interstitium were co-stained with CD34 or alpha-SMA, suggesting that peritubular endothelial cells and tubulointerstitial myofibroblasts express nestin during the progression of tubulointerstitial injury. In addition, strong nestin expression was associated with deterioration of renal function. CONCLUSION: Nestin expression is associated with tubulointerstitial injury and predicts renal prognosis in IgA nephropathy. Nestin could be a new marker for peritubular endothelial cell injury and tubulointerstitial fibrosis.


Assuntos
Glomerulonefrite por IGA/metabolismo , Proteínas de Filamentos Intermediários/análise , Túbulos Renais/química , Proteínas do Tecido Nervoso/análise , Actinas/análise , Adulto , Antígenos CD34/análise , Biomarcadores/análise , Biomarcadores/sangue , Biópsia , Creatinina/sangue , Feminino , Fibrose , Imunofluorescência , Taxa de Filtração Glomerular , Glomerulonefrite por IGA/patologia , Glomerulonefrite por IGA/fisiopatologia , Humanos , Imuno-Histoquímica , Japão , Estimativa de Kaplan-Meier , Túbulos Renais/patologia , Túbulos Renais/fisiopatologia , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Nestina , Valor Preditivo dos Testes , Prognóstico , Proteinúria/metabolismo , Processamento de Sinais Assistido por Computador , Fatores de Tempo
5.
Nephrol Dial Transplant ; 24(8): 2378-83, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19188342

RESUMO

BACKGROUND: Recent studies have reported that statins have renoprotective effects, independent from lowering plasma cholesterol. In this study, we examined whether statins were beneficial in a murine model of HIV-associated nephropathy (HIVAN). METHODS: We used conditional transgenic mice that express one of the HIV-1 accessory genes, vpr, selectively in podocytes using podocin promoter and the Tet-on system. These mice develop aggressive collapsing focal segmental glomerular sclerosis with massive proteinuria and deterioration of renal function within 4 weeks following heminephrectomy and doxycycline administration. Fluvastatin was administrated simultaneously with doxycycline, and the effect was compared with untreated controls after 4 weeks. RESULTS: Fluvastatin at 10 mg/kg/day significantly decreased urinary albumin excretion (87 versus 11 mg/day, P < 0.01) and glomerular sclerosis (2.4 versus 1.0, P < 0.01, assessed by semi-quantitative scoring: 0-4). Fluvastatin also decreased serum creatinine and total cholesterol, but these differences were not statistically significant (0.36 versus 0.32 mg/dl, P = 0.35; 492 versus 378 mg/dl, P = 0.11, respectively). Phenotypic changes in podocytes, as indicated by the downregulation of nephrin, Wilms' tumour 1 and synaptopodin, along with upregulation of proliferating cell nuclear antigen, were attenuated by fluvastatin, suggesting its protective effects against podocyte injuries. In cultured podocytes, angiotensin II treatment decreased nephrin expression to 13% of basal levels, which was reversed to 58% by adding fluvastatin. CONCLUSIONS: In conclusion, fluvastatin was effective in treating experimental HIVAN. The beneficial effect of this drug might be caused, in part, by preserving nephrin expression in podocytes against angiotensin II-mediated injury.


Assuntos
Nefropatia Associada a AIDS/prevenção & controle , Bloqueadores do Receptor Tipo 1 de Angiotensina II/uso terapêutico , Modelos Animais de Doenças , Ácidos Graxos Monoinsaturados/uso terapêutico , Glomerulosclerose Segmentar e Focal/prevenção & controle , Indóis/uso terapêutico , Podócitos/efeitos dos fármacos , Proteinúria/prevenção & controle , Nefropatia Associada a AIDS/patologia , Angiotensina II/metabolismo , Animais , Western Blotting , Células Cultivadas , Imunofluorescência , Fluvastatina , Genes do Tumor de Wilms , Técnicas Imunoenzimáticas , Rim/citologia , Rim/metabolismo , Rim/cirurgia , Masculino , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Transgênicos , Proteínas dos Microfilamentos/metabolismo , Nefrectomia , Fenótipo , Podócitos/metabolismo , Podócitos/patologia
6.
Nihon Shokakibyo Gakkai Zasshi ; 105(3): 391-6, 2008 Mar.
Artigo em Japonês | MEDLINE | ID: mdl-18332604

RESUMO

A 56-year-old man, who had epigastralgia and abnormal liver function test, was admitted to our hospital. ERCP showed common bile duct dilation and stones. We therefore performed endoscopic lithotripsy after endoscopic sphincterotomy. Ten months later, MRCP showed recurrent CBD stones. After rehospitalization, resection of the dilated bile duct was carried out. This case was diagnosed Todani's type Ib congenital cystic dilatation of common bile duct. Todani's type Ib is rare type of congenital dilatation of bile duct, and is interesting embryologically.


Assuntos
Cisto do Colédoco/patologia , Colangiopancreatografia Retrógrada Endoscópica , Cisto do Colédoco/diagnóstico , Cisto do Colédoco/cirurgia , Ducto Colédoco/patologia , Ducto Colédoco/cirurgia , Dilatação Patológica , Endoscopia do Sistema Digestório , Humanos , Litotripsia , Masculino , Pessoa de Meia-Idade
8.
Am J Physiol Renal Physiol ; 293(4): F1214-21, 2007 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17652372

RESUMO

Conditional transgenic mice that express one of the human immunodeficiency virus (HIV)-1 accessory genes, vpr, selectively in podocytes using a podocin promoter and a tetracycline-inducible system develop renal injuries similar to those of patients with HIV-associated nephropathy (HIVAN). We have shown that a heminephrectomy accelerates podocyte injury, which is alleviated by angiotensin II (ANG II) type 1 receptor blocker (ARB). The current study further explores the role of ANG II in the genesis of HIVAN in this murine model. With ANG II infusion, heavy proteinuria was observed at 1 wk after the initiation of doxycycline administration to induce vpr expression in podocytes. Severe morphological and phenotypical changes in the podocytes were observed at 2 wk, together with extensive glomerulosclerosis. Norepinephrine infusion, instead of ANG II, increased the systemic blood pressure to the same level as that achieved using ANG II. However, albuminuria and glomerular injury were modest in norepinephrine-infused mice. Treatment with an ARB, olmesartan, almost completely inhibited glomerular injury. In contrast, lowering the blood pressure with a vasodilator, hydralazine, partially decreased albuminuria but did not produce any histological changes. ANG II infusion alone without doxycycline resulted in a lower level of albuminuria and minimal histological changes. These data demonstrate that excessive ANG II accelerates vpr-induced podocyte injury in a mouse model of HIVAN.


Assuntos
Nefropatia Associada a AIDS/patologia , Angiotensina II/fisiologia , Podócitos/efeitos dos fármacos , Podócitos/patologia , Nefropatia Associada a AIDS/metabolismo , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Animais , Anti-Hipertensivos/farmacologia , Pressão Sanguínea/efeitos dos fármacos , Pressão Sanguínea/fisiologia , Modelos Animais de Doenças , Produtos do Gene vpr/genética , Produtos do Gene vpr/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Transgênicos , Norepinefrina/farmacologia , Fenótipo , Podócitos/metabolismo , Receptor Tipo 1 de Angiotensina/efeitos dos fármacos , Receptor Tipo 1 de Angiotensina/fisiologia
9.
J Am Soc Nephrol ; 18(2): 515-27, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17229913

RESUMO

HIV-associated nephropathy (HIVAN) is characterized by a collapsed glomerular capillary tuft with hyperplasia and hypertrophy of podocytes. Recently generated were conditional transgenic mice (podocin/Vpr) that express one of the HIV-1 accessory genes, vpr, selectively in podocytes using podocin promoter and Tet-on system. These transgenic mice developed renal injury similar to HIVAN when treated with doxycycline for 8 to 12 wk. This study demonstrated that nephron reduction by heminephrectomy markedly enhanced phenotypic changes of podocytes and led to severe FSGS within 4 wk. Nephrotic-range proteinuria was observed already at 2 wk, together with dedifferentiation and dysregulation of podocytes, indicated by decreased expression of nephrin, synaptopodin, and Wilms' tumor 1 protein and increased expression of Ki-67. The acceleration of phenotypic changes of podocytes, proteinuria, and subsequent glomerulosclerosis by heminephrectomy was almost completely inhibited by angiotensin II type 1 receptor (AT1R) blocker olmesartan. In contrast, the renoprotective effect of the calcium channel antagonist azelnidipine was minimal, although it lowered systemic BP to the same level as olmesartan, demonstrating that the inhibitory effect of AT1R blocker was independent of systemic BP. Olmesartan also reduced proteinuria and prevented glomerulosclerosis even by the delayed treatment, which was initiated after the podocyte injury appeared. These data suggest that nephron reduction exaggerates podocyte injury and subsequent glomerulosclerosis, possibly through glomerular hypertension, in the mouse model of HIVAN. AT1R blockade could be beneficial in the treatment of HIVAN by ameliorating podocyte injury by avoiding the vicious cycle of nephron reduction and glomerular hypertension.


Assuntos
Nefropatia Associada a AIDS/prevenção & controle , Bloqueadores do Receptor Tipo 1 de Angiotensina II/uso terapêutico , Receptor Tipo 1 de Angiotensina/fisiologia , Nefropatia Associada a AIDS/patologia , Animais , População Negra , Modelos Animais de Doenças , Humanos , Imidazóis/uso terapêutico , Peptídeos e Proteínas de Sinalização Intracelular/genética , Rim/patologia , Falência Renal Crônica/etiologia , Proteínas de Membrana/genética , Camundongos , Camundongos Transgênicos , Receptor Tipo 1 de Angiotensina/efeitos dos fármacos , Tetrazóis/uso terapêutico
10.
Jpn J Cancer Res ; 93(7): 774-82, 2002 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12149143

RESUMO

We examined in the present study whether human gastric cancer cells express peroxisome proliferator-activated receptor gamma (PPARgamma), the effect of PPARgamma activation by troglitazone, a selective ligand, on cellular growth, and the mechanism of the growth arrest by troglitazone in gastric cancer cells. RT-PCR, northern blot and western blot analysis demonstrated that all four tested human gastric cancer cell lines, MKN-28, MKN-45, MKN-74 and KATO-III, expressed PPARgamma mRNA and protein. WST-1 assay and flow cytometric analysis revealed that troglitazone inhibited the growth and induced G1 arrest in all four gastric cancer cell lines. To examine the role of p27(Kip1), a cyclin-dependent kinase inhibitor, in the G1 arrest by troglitazone, we determined p27(Kip1) protein expression by western blot analysis in gastric cancer cells that had been treated with troglitazone. Troglitazone increased p27(Kip1) in all four gastric cancer cell lines. Since it has been reported that the ubiquitin-proteasome system plays a vital role in the degradation of p27(Kip1) protein, we evaluated the hypothesis that inhibition of proteasome mediates the troglitazone-induced p27(Kip1) accumulation. Lactacystin, a proteasome inhibitor, inhibited cell growth and increased p27(Kip1) expression in MKN-74 cells. It was further demonstrated that troglitazone inhibited proteasome activity in a dose-dependent manner in MKN-74 cells. All these results suggest that troglitazone inhibited proteasome activity, followed by induction of p27(Kip1), which arrests cells at the G1 phase of the cell cycle in gastric cancer cells. The troglitazone-mediated inhibition of the proteasome suggests a novel mechanism for the anti-proliferative effect of this agent in cancer cells.


Assuntos
Proteínas de Ciclo Celular/metabolismo , Cromanos/farmacologia , Fase G1 , Complexos Multienzimáticos/antagonistas & inibidores , Neoplasias Gástricas/metabolismo , Tiazóis/farmacologia , Tiazolidinedionas , Proteínas Supressoras de Tumor/metabolismo , Antineoplásicos/farmacologia , Northern Blotting , Western Blotting , Ciclo Celular , Divisão Celular , Inibidor de Quinase Dependente de Ciclina p27 , Cisteína Endopeptidases , Inibidores Enzimáticos/farmacologia , Citometria de Fluxo , Humanos , Ligantes , Complexo de Endopeptidases do Proteassoma , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , RNA/metabolismo , RNA Mensageiro/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Fatores de Transcrição/metabolismo , Troglitazona , Células Tumorais Cultivadas
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