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1.
Hepatology ; 75(6): 1523-1538, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-34773257

RESUMO

BACKGROUND AND AIMS: Currently there is no Food and Drug Administration-approved drug to treat NAFLD and NASH, the rates of which are increasing worldwide. Although NAFLD/NASH are highly complex and heterogeneous conditions, most pharmacotherapy pipelines focus on a single mechanistic target. Considering the importance of the gut-liver axis in their pathogenesis, we investigated the therapeutic effect of a long-acting dual agonist of glucagon-like peptide (GLP)-1 and GLP-2 receptors in mice with NAFLD/NASH. APPROACH AND RESULTS: C57BL/6J mice were fed a choline-deficient high-fat diet/high fructose and sucrose solution. After 16 weeks, mice were randomly allocated to receive vehicle, GLP1-Fc, GLP2-Fc, or GLP1/2-Fc fusion (GLP1/2-Fc) subcutaneously every 2 days for 4 weeks. Body weight was monitored, insulin/glucose tolerance tests were performed, feces were collected, and microbiome profiles were analyzed. Immobilized cell systems were used to evaluate direct peptide effect. Immunohistochemistry, quantitative PCR, immunoblot analysis, tunnel assay, and biochemical assays were performed to assess drug effects on inflammation, hepatic fibrosis, cell death, and intestinal structures. The mice had well-developed NASH phenotypes. GLP1/2-Fc reduced body weight, glucose levels, hepatic triglyceride levels, and cellular apoptosis. It improved liver fibrosis, insulin sensitivity, and intestinal tight junctions, and increased microvillus height, crypt depth, and goblet cells of intestine compared with a vehicle group. Similar effects of GLP1/2-Fc were found in in vitro cell systems. GLP1/2-Fc also changed microbiome profiles. We applied fecal microbiota transplantation (FMT) gain further insight into the mechanism of GLP1/2-Fc-mediated protection. We confirmed that FMT exerted an additive effect on GLP1-Fc group, including the body weight change, liver weight, hepatic fat accumulation, inflammation, and hepatic fibrosis. CONCLUSIONS: A long-acting dual agonist of GLP-1 and GLP-2 receptors is a promising therapeutic strategy to treat NAFLD/NASH.


Assuntos
Microbiota , Hepatopatia Gordurosa não Alcoólica , Animais , Peso Corporal , Dieta Hiperlipídica/efeitos adversos , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Receptor do Peptídeo Semelhante ao Glucagon 2/metabolismo , Inflamação/metabolismo , Fígado/patologia , Cirrose Hepática/complicações , Camundongos , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/patologia
2.
J Korean Med Sci ; 37(37): e276, 2022 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-36163475

RESUMO

Autophagy is critically involved in the maintenance of intracellular nutrient homeostasis and organelle function. Dysregulated autophagy is likely to play a role in the development of metabolic disorders and diabetes because autophagy is critical in the rejuvenation of dysfunctional or stressed endoplasmic reticulum and mitochondria that play a crucial role in the development of diabetes. Indeed, systemic autophagy insufficiency led to the increased tissue lipid content, aggravated metabolic and finally more severe diabetes when metabolic stress was imposed, suggesting that autophagy insufficiency of dysfunction of lysosome, an effector organelle of autophagy, due to aging, genetic predisposition or environmental factors could be an underlying cause of diabetes. Conversely, autophagy enhancer could improve metabolic profile of obese mice by reducing tissue lipid content and ameliorating metabolic inflammation. Furthermore, clearance of human islet amyloid polypeptide (hIAPP) oligomer and amyloid that accumulate in pancreatic islets of > 90% of diabetes patients was also dependent on autophagy. Consistently, autophagy enhancer could improve glucose profile and ß-cell function of transgenic mice expressing amyloidogenic hIAPP in pancreatic ß-cells, which was accompanied by reduced accumulation of hIAPP oligomer or amyloid, ameliorated ß-cell apoptosis and increased ß-cell mass. These results suggest that autophagy enhancer could be a novel therapeutic modality against diabetes associated with lipid overload and human diabetes characterized by islet amyloid accumulation.


Assuntos
Diabetes Mellitus Tipo 2 , Diabetes Mellitus , Células Secretoras de Insulina , Ilhotas Pancreáticas , Síndrome Metabólica , Amiloide/genética , Amiloide/metabolismo , Animais , Autofagia/fisiologia , Diabetes Mellitus/tratamento farmacológico , Diabetes Mellitus/patologia , Diabetes Mellitus Tipo 2/patologia , Glucose/metabolismo , Humanos , Células Secretoras de Insulina/metabolismo , Polipeptídeo Amiloide das Ilhotas Pancreáticas/química , Polipeptídeo Amiloide das Ilhotas Pancreáticas/genética , Polipeptídeo Amiloide das Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/metabolismo , Ilhotas Pancreáticas/patologia , Lipídeos , Síndrome Metabólica/complicações , Camundongos , Camundongos Transgênicos
3.
Mol Cell ; 51(5): 618-31, 2013 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-24011591

RESUMO

The Keap1-Nrf2 system and autophagy are both involved in the oxidative-stress response, metabolic pathways, and innate immunity, and dysregulation of these processes is associated with pathogenic processes. However, the interplay between these two pathways remains largely unknown. Here, we show that phosphorylation of the autophagy-adaptor protein p62 markedly increases p62's binding affinity for Keap1, an adaptor of the Cul3-ubiquitin E3 ligase complex responsible for degrading Nrf2. Thus, p62 phosphorylation induces expression of cytoprotective Nrf2 targets. p62 is assembled on selective autophagic cargos such as ubiquitinated organelles and subsequently phosphorylated in an mTORC1-dependent manner, implying coupling of the Keap1-Nrf2 system to autophagy. Furthermore, persistent activation of Nrf2 through accumulation of phosphorylated p62 contributes to the growth of human hepatocellular carcinomas (HCCs). These results demonstrate that selective autophagy and the Keap1-Nrf2 pathway are interdependent, and that inhibitors of the interaction between phosphorylated p62 and Keap1 have potential as therapeutic agents against human HCC.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/química , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Autofagia/fisiologia , Proteínas do Citoesqueleto/química , Proteínas do Citoesqueleto/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Adenoma/metabolismo , Adenoma/patologia , Sequência de Aminoácidos , Animais , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , Cristalografia por Raios X , Proteínas de Choque Térmico/química , Proteínas de Choque Térmico/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Alvo Mecanístico do Complexo 1 de Rapamicina , Camundongos , Dados de Sequência Molecular , Complexos Multiproteicos/metabolismo , Fosforilação , Proteína Sequestossoma-1 , Serina-Treonina Quinases TOR/metabolismo
4.
Biochem Soc Trans ; 48(3): 1213-1225, 2020 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-32510139

RESUMO

The importance of innate immunity in host defense and inflammatory responses has been clearly demonstrated after the discovery of innate immune receptors such as Toll-like receptors (TLRs) or Nucleotide-binding oligomerization domain-containing protein (Nod)-like receptors (NLRs). Innate immunity also plays a critical role in diverse pathological conditions including autoimmune diseases such as type 1 diabetes (T1D). In particular, the role of a variety of innate immune receptors in T1D has been demonstrated using mice with targeted disruption of such innate immune receptors. Here, we discuss recent findings showing the role of innate immunity in T1D that were obtained mostly from studies of genetic mouse models of innate immune receptors. In addition, the role of innate immune receptors involved in the pathogenesis of T1D in sensing death-associated molecular patterns (DAMPs) released from dead cells or pathogen-associated molecular patterns (PAMPs) will also be covered. Elucidation of the role of innate immune receptors in T1D and the nature of DAMPs sensed by such receptors may lead to the development of new therapeutic modalities against T1D.


Assuntos
Morte Celular , Diabetes Mellitus Tipo 1/imunologia , Imunidade Inata , Moléculas com Motivos Associados a Patógenos/imunologia , Animais , Células Dendríticas/metabolismo , Humanos , Inflamação , Ligantes , Macrófagos/metabolismo , Complexo Principal de Histocompatibilidade , Camundongos , Camundongos Endogâmicos NOD , Modelos Genéticos , Fator 88 de Diferenciação Mieloide/metabolismo , Nucleotídeos/metabolismo , Receptores Imunológicos/metabolismo , Transdução de Sinais/imunologia , Receptores Toll-Like/metabolismo
5.
Biochem Biophys Res Commun ; 508(3): 965-972, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30545632

RESUMO

Renal fibrosis is the final common pathway of various renal injuries and it leads to chronic kidney disease. Recent studies reported that FOXD1-lineage pericyte plays a critical role in tubulointerstitial fibrosis (TIF). However the regulatory mechanisms remain unclear. Autophagy is a cellular process of degradation of damaged cytoplasmic components that regulates cell death and proliferation. To investigate the role of autophagy in FOXD1-lineage pericytes on renal TIF, we generated the FOXD1-lineage stromal cell-specific Atg7 deletion (Atg7△FOXD1) mice. FOXD1-lineage stromal cell-specific Atg7 deletion enhanced renal TIF through Smad-dependent transforming growth factor (TGF)-ß signaling after unilateral ureteral obstruction (UUO). FOXD1-lineage stromal cell-specific Atg7 deletion increased the accumulation of interstitial myofibroblasts and enhanced the differentiation of pericytes into myofibroblasts after UUO. Peritubular capillary rarefaction was accelerated in Atg7△FOXD1 mice after UUO. Atg7△FOXD1 mice increased the accumulation of SQSTM1/p62-positive aggregates in the obstructed kidney and resulted in increased expression of NLRP3 inflammasome, interleukin (IL) 1-ß and caspase-1 signaling pathway, which enhanced apoptosis of interstitial cells after UUO. In summary, our data showed that autophagy in FOXD1-lineage stromal cells plays a protective role in renal TIF through regulating the Smad4 dependent TGF-ß an NLRP3 inflammasome signaling pathway.


Assuntos
Autofagia , Fatores de Transcrição Forkhead/análise , Inflamassomos/metabolismo , Rim/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Animais , Apoptose , Proteína 7 Relacionada à Autofagia/genética , Diferenciação Celular , Linhagem da Célula , Fibrose , Fatores de Transcrição Forkhead/genética , Rim/citologia , Rim/metabolismo , Masculino , Camundongos , Camundongos Knockout , Miofibroblastos/citologia , Pericitos/citologia , Receptores do Fator de Crescimento Derivado de Plaquetas/análise , Transdução de Sinais , Proteínas Smad/metabolismo , Células Estromais/química , Obstrução Ureteral/complicações
6.
Am J Hum Genet ; 96(2): 266-74, 2015 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-25620203

RESUMO

Singleton-Merten syndrome (SMS) is an autosomal-dominant multi-system disorder characterized by dental dysplasia, aortic calcification, skeletal abnormalities, glaucoma, psoriasis, and other conditions. Despite an apparent autosomal-dominant pattern of inheritance, the genetic background of SMS and information about its phenotypic heterogeneity remain unknown. Recently, we found a family affected by glaucoma, aortic calcification, and skeletal abnormalities. Unlike subjects with classic SMS, affected individuals showed normal dentition, suggesting atypical SMS. To identify genetic causes of the disease, we performed exome sequencing in this family and identified a variant (c.1118A>C [p.Glu373Ala]) of DDX58, whose protein product is also known as RIG-I. Further analysis of DDX58 in 100 individuals with congenital glaucoma identified another variant (c.803G>T [p.Cys268Phe]) in a family who harbored neither dental anomalies nor aortic calcification but who suffered from glaucoma and skeletal abnormalities. Cys268 and Glu373 residues of DDX58 belong to ATP-binding motifs I and II, respectively, and these residues are predicted to be located closer to the ADP and RNA molecules than other nonpathogenic missense variants by protein structure analysis. Functional assays revealed that DDX58 alterations confer constitutive activation and thus lead to increased interferon (IFN) activity and IFN-stimulated gene expression. In addition, when we transduced primary human trabecular meshwork cells with c.803G>T (p.Cys268Phe) and c.1118A>C (p.Glu373Ala) mutants, cytopathic effects and a significant decrease in cell number were observed. Taken together, our results demonstrate that DDX58 mutations cause atypical SMS manifesting with variable expression of glaucoma, aortic calcification, and skeletal abnormalities without dental anomalies.


Assuntos
Doenças da Aorta/genética , RNA Helicases DEAD-box/genética , Hipoplasia do Esmalte Dentário/genética , Glaucoma/genética , Metacarpo/anormalidades , Modelos Moleculares , Doenças Musculares/genética , Odontodisplasia/genética , Osteoporose/genética , Calcificação Vascular/genética , Adulto , Doenças da Aorta/patologia , Sequência de Bases , Células Cultivadas , Pré-Escolar , Proteína DEAD-box 58 , RNA Helicases DEAD-box/química , Hipoplasia do Esmalte Dentário/patologia , Exoma/genética , Feminino , Genes Dominantes/genética , Humanos , Masculino , Metacarpo/patologia , Dados de Sequência Molecular , Doenças Musculares/patologia , Anormalidades Musculoesqueléticas/diagnóstico por imagem , Anormalidades Musculoesqueléticas/genética , Mutação de Sentido Incorreto/genética , Odontodisplasia/diagnóstico por imagem , Odontodisplasia/patologia , Osteoporose/patologia , Linhagem , Polimorfismo de Nucleotídeo Único/genética , Radiografia , Receptores Imunológicos , Análise de Sequência de DNA , Calcificação Vascular/patologia
7.
Hum Mol Genet ; 24(25): 7196-206, 2015 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-26433933

RESUMO

Really interesting new gene (RING) finger protein 170 (RNF170) is an E3 ubiquitin ligase known to mediate ubiquitination-dependent degradation of type-I inositol 1,4,5-trisphosphate receptors (ITPR1). It has recently been demonstrated that a point mutation of RNF170 gene is linked with autosomal-dominant sensory ataxia (ADSA), which is characterized by an age-dependent increase of walking abnormalities, a rare genetic disorder reported in only two families. Although this mutant allele is known to be dominant, the functional identity thereof has not been clearly established. Here, we generated mice lacking Rnf170 (Rnf170(-/-)) to evaluate the effect of its loss of function in vivo. Remarkably, Rnf170(-/-) mice began to develop gait abnormalities in old age (12 months) in the form of asynchronous stepping between diagonal limb pairs with a fixed step sequence during locomotion, while age-matched wild-type mice showed stable gait patterns using several step sequence repertoires. As reported in ADSA patients, they also showed a reduced sensitivity for proprioception and thermal nociception. Protein blot analysis revealed that the amount of Itpr1 protein was significantly elevated in the cerebellum and spinal cord but intact in the cerebral cortex in Rnf170(-/-) mice. These results suggest that the loss of Rnf170 gene function mediates ADSA-associated phenotypes and this gives insights on the cure of patients with ADSA and other age-dependent walking abnormalities.


Assuntos
Ataxia/congênito , Marcha/fisiologia , Ubiquitina-Proteína Ligases/genética , Fatores Etários , Animais , Ataxia/genética , Ataxia/fisiopatologia , Marcha/genética , Humanos , Camundongos , Camundongos Knockout , Ubiquitina-Proteína Ligases/deficiência
8.
Biochem J ; 473(12): 1791-803, 2016 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-27095850

RESUMO

Secretagogin (SCGN), a Ca(2+)-binding protein having six EF-hands, is selectively expressed in pancreatic ß-cells and neuroendocrine cells. Previous studies suggested that SCGN enhances insulin secretion by functioning as a Ca(2+)-sensor protein, but the underlying mechanism has not been elucidated. The present study explored the mechanism by which SCGN enhances glucose-induced insulin secretion in NIT-1 insulinoma cells. To determine whether SCGN influences the first or second phase of insulin secretion, we examined how SCGN affects the kinetics of insulin secretion in NIT-1 cells. We found that silencing SCGN suppressed the second phase of insulin secretion induced by glucose and H2O2, but not the first phase induced by KCl stimulation. Recruitment of insulin granules in the second phase of insulin secretion was significantly impaired by knocking down SCGN in NIT-1 cells. In addition, we found that SCGN interacts with the actin cytoskeleton in the plasma membrane and regulates actin remodelling in a glucose-dependent manner. Since actin dynamics are known to regulate focal adhesion, a critical step in the second phase of insulin secretion, we examined the effect of silencing SCGN on focal adhesion molecules, including FAK (focal adhesion kinase) and paxillin, and the cell survival molecules ERK1/2 (extracellular-signal-regulated kinase 1/2) and Akt. We found that glucose- and H2O2-induced activation of FAK, paxillin, ERK1/2 and Akt was significantly blocked by silencing SCGN. We conclude that SCGN controls glucose-stimulated insulin secretion and thus may be useful in the therapy of Type 2 diabetes.


Assuntos
Actinas/metabolismo , Adesões Focais/metabolismo , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Secretagoginas/metabolismo , Animais , Western Blotting , Linhagem Celular Tumoral , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Adesões Focais/efeitos dos fármacos , Adesões Focais/ultraestrutura , Glucose/farmacologia , Peróxido de Hidrogênio/farmacologia , Imunoprecipitação , Secreção de Insulina , Células Secretoras de Insulina/efeitos dos fármacos , Camundongos , Microscopia Confocal , Microscopia Eletrônica de Transmissão , Paxilina/metabolismo , Cloreto de Potássio/farmacologia , Ligação Proteica , Secretagoginas/genética
9.
Diabetologia ; 59(7): 1480-1491, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26831301

RESUMO

AIMS/HYPOTHESIS: We studied the role of protein degradation pathways in the regulation of insulin production and secretion and hypothesised that autophagy regulates proinsulin degradation, thereby modulating beta cell function. METHODS: Proinsulin localisation in autophagosomes was demonstrated by confocal and electron microscopy. Autophagy was inhibited by knockdown of autophagy-related (ATG) proteins and using the H(+)-ATPase inhibitor bafilomycin-A1. Proinsulin and insulin content and secretion were assessed in static incubations by ELISA and RIA. RESULTS: Confocal and electron microscopy showed proinsulin localised in autophagosomes and lysosomes. Beta-Atg7 (-/-) mice had proinsulin-containing sequestosome 1 (p62 [also known as SQSTM1])(+) aggregates in beta cells, indicating proinsulin is regulated by autophagy in vivo. Short-term bafilomycin-A1 treatment and ATG5/7 knockdown increased steady-state proinsulin and hormone precursor chromogranin A content. ATG5/7 knockdown also increased glucose- and non-fuel-stimulated insulin secretion. Finally, mutated forms of proinsulin that are irreparably misfolded and trapped in the endoplasmic reticulum are more resistant to degradation by autophagy. CONCLUSIONS/INTERPRETATION: In the beta cell, transport-competent secretory peptide precursors, including proinsulin, are regulated by autophagy, whereas efficient clearance of transport-incompetent mutated forms of proinsulin by alternative degradative pathways may be necessary to avoid beta cell proteotoxicity. Reduction of autophagic degradation of proinsulin increases its residency in the secretory pathway, followed by enhanced secretion in response to stimuli.


Assuntos
Autofagia/fisiologia , Insulina/metabolismo , Animais , Autofagia/genética , Proteína 7 Relacionada à Autofagia/genética , Proteína 7 Relacionada à Autofagia/metabolismo , Western Blotting , Linhagem Celular , Homeostase/genética , Homeostase/fisiologia , Humanos , Células Secretoras de Insulina/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Eletrônica , Consumo de Oxigênio/genética , Consumo de Oxigênio/fisiologia , Interferência de RNA/fisiologia
10.
Diabetologia ; 58(4): 809-18, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25537833

RESUMO

AIMS/HYPOTHESIS: Fibroblast growth factor 21 (FGF21) is an endocrine hormone that exhibits anti-diabetic and anti-obesity activity. FGF21 expression is increased in patients with and mouse models of obesity or nonalcoholic fatty liver disease (NAFLD). However, the functional role and molecular mechanism of FGF21 induction in obesity or NAFLD are not clear. As endoplasmic reticulum (ER) stress is triggered in obesity and NAFLD, we investigated whether ER stress affects FGF21 expression or whether FGF21 induction acts as a mechanism of the unfolded protein response (UPR) adaptation to ER stress induced by chemical stressors or obesity. METHODS: Hepatocytes or mouse embryonic fibroblasts deficient in UPR signalling pathways and liver-specific eIF2α mutant mice were employed to investigate the in vitro and in vivo effects of ER stress on FGF21 expression, respectively. The in vivo importance of FGF21 induction by ER stress and obesity was determined using inducible Fgf21-transgenic mice and Fgf21-null mice with or without leptin deficiency. RESULTS: We found that ER stressors induced FGF21 expression, which was dependent on a PKR-like ER kinase-eukaryotic translation factor 2α-activating transcription factor 4 pathway both in vitro and in vivo. Fgf21-null mice exhibited increased expression of ER stress marker genes and augmented hepatic lipid accumulation after tunicamycin treatment. However, these changes were attenuated in inducible Fgf21-transgenic mice. We also observed that Fgf21-null mice with leptin deficiency displayed increased hepatic ER stress response and liver injury, accompanied by deteriorated metabolic variables. CONCLUSIONS/INTERPRETATION: Our results suggest that FGF21 plays an important role in the adaptive response to ER stress- or obesity-induced hepatic metabolic stress.


Assuntos
Estresse do Retículo Endoplasmático , Retículo Endoplasmático/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Hepatócitos/metabolismo , Obesidade/metabolismo , Estresse Fisiológico , Fator 4 Ativador da Transcrição/metabolismo , Adaptação Fisiológica , Animais , Modelos Animais de Doenças , Fator de Iniciação 2 em Eucariotos/metabolismo , Fatores de Crescimento de Fibroblastos/deficiência , Fatores de Crescimento de Fibroblastos/genética , Células Hep G2 , Humanos , Camundongos Knockout , Camundongos Obesos , Obesidade/genética , Obesidade/fisiopatologia , Interferência de RNA , Transdução de Sinais , Fatores de Tempo , Transfecção , Resposta a Proteínas não Dobradas , eIF-2 Quinase/metabolismo
11.
J Biol Chem ; 289(36): 24944-55, 2014 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-25049227

RESUMO

The ubiquitin-proteasome system and autophagy are crucially important for proteostasis in cells. These pathways are interdependent, and dysfunction in either pathway causes accumulation of ubiquitin-positive aggregates, a hallmark of human pathological conditions. To elucidate in vivo compensatory action(s) against proteasomal dysfunction, we developed mice with reduced proteasome activity in their livers. The mutant mice exhibited severe liver damage, accompanied by formation of aggregates positive for ubiquitin and p62/Sqstm1, an adaptor protein for both selective autophagy and the anti-oxidative Keap1-Nrf2 pathway. These aggregates were selectively entrapped by autophagosomes, and pathological features of livers with impaired proteasome activity were exacerbated by simultaneous suppression of autophagy. In contrast, concomitant loss of p62/Sqstm1 had no apparent effect on the liver pathology though p62/Sqstm1 was indispensable for the aggregates formation. Furthermore, defective proteasome function led to transcriptional activation of the Nrf2, which served as a physiological adaptation. Our in vivo data suggest that cells contain networks of cellular defense mechanisms against defective proteostasis.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Autofagia , Proteínas do Citoesqueleto/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Complexo de Endopeptidases do Proteassoma/metabolismo , Transdução de Sinais , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Proteínas do Citoesqueleto/genética , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Immunoblotting , Proteína 1 Associada a ECH Semelhante a Kelch , Fígado/metabolismo , Fígado/patologia , Fígado/ultraestrutura , Camundongos Knockout , Camundongos Transgênicos , Microscopia Confocal , Microscopia Imunoeletrônica , Fator 2 Relacionado a NF-E2/genética , Fagossomos/genética , Fagossomos/metabolismo , Fosforilação , Complexo de Endopeptidases do Proteassoma/genética , Proteína Sequestossoma-1 , Fatores de Tempo , Ubiquitina/metabolismo
12.
Cardiovasc Diabetol ; 14: 70, 2015 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-26041130

RESUMO

BACKGROUND: It is presently unclear whether glycemic variability is associated with diabetic cardiovascular autonomic neuropathy (CAN). The aim of this study was to examine whether short- and/or long-term glycemic variability (GV) contribute to CAN. METHODS: A total of 110 patients with type 2 diabetes who underwent three-day continuous glucose monitoring (CGM) completed five standardized autonomic neuropathy tests. Short-term GV was measured by the standard deviation (SD), coefficient of variation (CV) of glucose, and the mean amplitude of glycemic excursions (MAGE) in CGM. HbA1c variability was calculated from the intrapersonal SD, adjusted SD, and CV of serial HbA1c over 2-year period. CAN was defined as the presence of at least two abnormal parasympathetic function tests. The severity of CAN was evaluated by total scores of five autonomic function tests. RESULTS: In univariate analysis, not only SD and CV in CGM but also all parameters of HbA1c variability were significantly higher in the patients with CAN (n = 47, 42.7 %) than in those without CAN. In multivariate analysis, CV (Odds ratio [OR] 1.07, 95 % confidence interval [CI] 1.01-1.13; p = 0.033), but neither SD nor MAGE in CGM, independently correlated with the presence of CAN. All parameters of HbA1c variability, such as SD of HbA1c (OR 12.10 [95 % CI 2.29-63.94], p = 0.003), adjusted SD of HbA1c (OR 17.02 [95 % CI 2.66-108.86], p = 0.003), and log CV of HbA1c (OR 24.00 [95 % CI 3.09-186.48], p = 0.002), were significantly associated with the presence of CAN. The patients with higher HbA1c variability had an increased risk of advanced CAN. CONCLUSION: CV in CGM and all parameters of HbA1c variability were independently associated with the presence of CAN in patients with inadequately controlled type 2 diabetes requiring CGM.


Assuntos
Doenças do Sistema Nervoso Autônomo/fisiopatologia , Glicemia/metabolismo , Doenças Cardiovasculares/fisiopatologia , Sistema Cardiovascular/inervação , Diabetes Mellitus Tipo 2/metabolismo , Neuropatias Diabéticas/fisiopatologia , Idoso , Doenças do Sistema Nervoso Autônomo/etiologia , Pressão Sanguínea/fisiologia , Doenças Cardiovasculares/etiologia , Estudos de Coortes , Diabetes Mellitus Tipo 2/complicações , Neuropatias Diabéticas/etiologia , Feminino , Hemoglobinas Glicadas/metabolismo , Frequência Cardíaca/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Monitorização Ambulatorial , Manobra de Valsalva
13.
J Immunol ; 190(12): 6368-77, 2013 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-23670194

RESUMO

Intracellular reactive oxygen species (ROS) are essential secondary messengers in many signaling cascades governing innate immunity and cellular functions. TLR3 signaling is crucially involved in antiviral innate and inflammatory responses; however, the roles of ROS in TLR3 signaling remain largely unknown. In this study, we show that TLR3-induced ROS generation is required for the activation of NF-κB, IFN-regulatory factor 3, and STAT1-mediated innate immune responses in macrophages. TLR3 induction led to a rapid increase in ROS generation and a physical association between components of the NADPH oxidase (NOX) enzyme complex (NOX2 and p47(phox)) and TLR3 via a Ca(2+)-c-Src tyrosine kinase-dependent pathway. TLR3-induced ROS generation, NOX2, and p47(phox) were required for the phosphorylation and nuclear translocation of STAT1 and STAT2. TLR3-induced activation of STAT1 contributed to the generation of inflammatory mediators, which was significantly attenuated in NOX2- and p47(phox)-deficient macrophages, suggesting a role for ROS-STAT1 in TLR3-mediated innate immune responses. Collectively, these results provide a novel insight into the crucial role that TLR3-ROS signaling plays in innate immune responses by activating STAT1.


Assuntos
Imunidade Inata/imunologia , Espécies Reativas de Oxigênio/imunologia , Fator de Transcrição STAT1/imunologia , Transdução de Sinais/imunologia , Receptor 3 Toll-Like/imunologia , Animais , Western Blotting , Células Cultivadas , Feminino , Imunofluorescência , Imunoprecipitação , Inflamação/imunologia , Inflamação/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Microscopia Confocal , Espécies Reativas de Oxigênio/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT1/metabolismo , Receptor 3 Toll-Like/metabolismo
14.
Endocr J ; 62(3): 243-50, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25476661

RESUMO

The aim of this study was to determine the diagnostic efficacy of free metanephrines in plasma samples drawn in the seated position compared with 24-h urinary metanephrines in detecting pheochromocytomas in Asian patients. This prospective study was conducted at Samsung Medical Center between May 2010 and July 2011. The study contained 245 subjects, including 28 patients with histologically-proven pheochromocytoma, 44 with histologically-proven non-pheochromocytoma, 112 controls suspected of having tumors but with negative investigations during two or more years of follow-up, and 45 healthy normotensive volunteers. Plasma-free metanephrines were measured by LC-MS/MS. The cut-off values with optimal sensitivity and specificity for plasma metanephrine and plasma normetanephrine were 0.33 nmol/L and 0.61 nmol/L, respectively. Both the plasma metanephrines measurement and urinary metanephrines measurement had a sensitivity of 96.4% (p = 1.00). However, the urinary metanephrines measurement was significantly more specific than the plasma metanephrines measurement (94.2% vs. 75.6%; p < 0.001). When we applied cut-off values based on BMI, specificity improved from 75.6% to 87.2%, with a comparable gain in sensitivity. From a diagnostic perspective, measurement of free metanephrines in plasma drawn in the seated position is highly sensitive but insufficiently specific when compared with measurement of 24-h urinary fractionated metanephrines. The specificity may be improved by applying cut-off values based on BMI. We suggest that free metanephrines in plasma drawn from seated position can also be used as an initial screening test to ensure that pheochromocytomas are not missed in Asian patients.


Assuntos
Neoplasias das Glândulas Suprarrenais/diagnóstico , Biomarcadores Tumorais/metabolismo , Metanefrina/metabolismo , Feocromocitoma/diagnóstico , Neoplasias das Glândulas Suprarrenais/sangue , Neoplasias das Glândulas Suprarrenais/urina , Adulto , Idoso , Biomarcadores Tumorais/sangue , Biomarcadores Tumorais/urina , Feminino , Humanos , Masculino , Metanefrina/sangue , Metanefrina/urina , Pessoa de Meia-Idade , Posicionamento do Paciente , Feocromocitoma/sangue , Feocromocitoma/urina , Estudos Prospectivos , Sensibilidade e Especificidade , Espectrometria de Massas em Tandem
15.
Gut ; 63(5): 727-35, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-23804561

RESUMO

BACKGROUND: Recent evidence indicates that the composition of the gut microbiota contributes to the development of metabolic disorders by affecting the physiology and metabolism of the host. Metformin is one of the most widely prescribed type 2 diabetes (T2D) therapeutic agents. OBJECTIVE: To determine whether the antidiabetic effect of metformin is related to alterations of intestinal microbial composition. DESIGN: C57BL/6 mice, fed either a normal-chow diet or a high-fat diet (HFD), were treated with metformin for 6 weeks. The effect of metformin on the composition of the gut microbiota was assessed by analysing 16S rRNA gene sequences with 454 pyrosequencing. Adipose tissue inflammation was examined by flow cytometric analysis of the immune cells present in visceral adipose tissue (VAT). RESULTS: Metformin treatment significantly improved the glycaemic profile of HFD-fed mice. HFD-fed mice treated with metformin showed a higher abundance of the mucin-degrading bacterium Akkermansia than HFD-fed control mice. In addition, the number of mucin-producing goblet cells was significantly increased by metformin treatment (p<0.0001). Oral administration of Akkermansia muciniphila to HFD-fed mice without metformin significantly enhanced glucose tolerance and attenuated adipose tissue inflammation by inducing Foxp3 regulatory T cells (Tregs) in the VAT. CONCLUSIONS: Modulation of the gut microbiota (by an increase in the Akkermansia spp. population) may contribute to the antidiabetic effects of metformin, thereby providing a new mechanism for the therapeutic effect of metformin in patients with T2D. This suggests that pharmacological manipulation of the gut microbiota in favour of Akkermansia may be a potential treatment for T2D.


Assuntos
Glicemia/efeitos dos fármacos , Homeostase/efeitos dos fármacos , Hipoglicemiantes/farmacologia , Intestinos/efeitos dos fármacos , Metformina/farmacologia , Microbiota/efeitos dos fármacos , Verrucomicrobia/crescimento & desenvolvimento , Animais , Biomarcadores/metabolismo , Glicemia/metabolismo , Dieta Hiperlipídica , Esquema de Medicação , Citometria de Fluxo , Hipoglicemiantes/administração & dosagem , Inflamação/imunologia , Intestinos/microbiologia , Gordura Intra-Abdominal/efeitos dos fármacos , Gordura Intra-Abdominal/imunologia , Metformina/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Obesos , Verrucomicrobia/isolamento & purificação
16.
J Hum Genet ; 59(9): 488-93, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25078357

RESUMO

Von Hippel-Lindau (VHL) disease is an inherited tumor syndrome caused by germline mutations in the VHL tumor suppressor gene. It is characterized by hemangioblastoma in the central nervous system and retina, renal cell carcinoma, pancreatic tumor and cysts, and pheochromocytoma. In this study, we detected 26 germline mutations in the VHL gene of Korean patients, of which 1 was a novel mutation, c.417_418insT. We also integrated our data from this study with the published literature to identify 55 VHL germline mutations in Koreans, and identified a unique hotspot at codon 70. Nine unrelated patients (9/55, 16.4%) had the same amino-acid substitution at codon 70 (Glu70Lys) and showed VHL type 1 phenotypes. Although this mutation was shown to have a mild effect on VHL function, four of the nine patients (44.4%) subsequently developed multiple central nervous system hemangioblastomas or retinal hemangioblastoma. However, this hotspot has not been identified in Chinese or Japanese patients. This study provides information on the spectrum of VHL mutations in Korean VHL disease and contributes to a better understanding of VHL disease in terms of improvements in the clinical management of VHL families.


Assuntos
Substituição de Aminoácidos , Mutação em Linhagem Germinativa , Proteína Supressora de Tumor Von Hippel-Lindau/genética , Doença de von Hippel-Lindau/genética , Adulto , Povo Asiático/genética , Criança , Análise Mutacional de DNA , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Fenótipo , República da Coreia , Estudos Retrospectivos , Adulto Jovem , Doença de von Hippel-Lindau/etnologia , Doença de von Hippel-Lindau/patologia
17.
Rev Endocr Metab Disord ; 15(1): 11-20, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24085381

RESUMO

Autophagy plays an important role in the regulation of cellular homeostasis through elimination of aggregated proteins, damaged organelles, and intracellular pathogens. Autophagy also contributes to the maintenance of energy balance through degradation of energy reserves such as lipids, glycogen, and proteins in the setting of increased energy demand. Recent studies have suggested that autophagy, or its deficiency, is implicated in the pathogenesis of insulin resistance, obesity, and diabetes. These effects of autophagy or its deficiency in regulation of energy metabolism are mediated not only by cell-autonomous effects, such as direct autophagic degradation of energy stores or intracellular organelles (endoplasmic reticulum and mitochondria) but also by non-cell-autonomous effects, such as induction/suppression of secreted factors or changes of sympathetic tone. In the present review, we highlight a recent surge in the research on the autophagy in the regulation of energy homeostasis, with a focus on its role as a mediator for crosstalk between metabolic organs.


Assuntos
Tecido Adiposo/metabolismo , Autofagia/fisiologia , Metabolismo Energético/fisiologia , Resistência à Insulina/fisiologia , Fígado/metabolismo , Músculo Esquelético/metabolismo , Animais , Fator 2 de Crescimento de Fibroblastos/metabolismo , Homeostase/fisiologia , Humanos , Interleucina-1beta/metabolismo , Obesidade/metabolismo
18.
Crit Rev Immunol ; 33(3): 245-81, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23756246

RESUMO

Type 1 diabetes is a prototypic, organ-specific autoimmune disease. Diverse antigen-specific immunotherapy using insulin or glutamic acid decarboxylase peptides and other immunotherapies, such as antibodies, fusion proteins, cytokines, regulatory T cells, small-molecule inhibitors, nonspecific immune modulators, or dietary modifications, have been attempted in human type 1 diabetes. Some of these immunotherapies delay the onset of diabetes or reduce insulin requirements or blood glucose level in patients with established type 1 diabetes. However, most of these immunotherapies failed to induce complete remission of established type 1 diabetes, which could be due to 1) technical difficulties in the achievement of immune tolerance to diabetic autoantigens or in the inhibition of autoimmune responses to those antigens that can be applied to human patients without significant adverse effects, and 2) markedly reduced ß-cell mass at the time of disease onset that should be replenished. This review focuses on the immunological aspects of the disease and its treatment, and data from previous or ongoing human clinical trials using immune-logical measures, and recent results from immunological studies employing animal models are discussed.


Assuntos
Diabetes Mellitus Tipo 1/terapia , Animais , Anticorpos/imunologia , Autoantígenos/imunologia , Ensaios Clínicos como Assunto , Diabetes Mellitus Tipo 1/imunologia , Humanos , Imunoterapia , Ilhotas Pancreáticas/imunologia
19.
J Korean Med Sci ; 29(8): 1038-41, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25120311

RESUMO

The importance of innate immunity in host defense is becoming clear after discovery of innate immune receptors such as Toll-like receptor or Nod-like receptor. Innate immune system plays an important role in diverse pathological situations such as autoimmune diseases. Role of innate immunity in the pathogenesis of metabolic disorders such as type 2 diabetes, metabolic syndrome or atherosclerosis that has not been previously considered as inflammatory disorders, is also being appreciated. Here, the role of innate immunity in the development of type 1 diabetes, a classical organ-specific autoimmune disease, and type 2 diabetes will be discussed, focusing on the role of specific innate immune receptors involved in these disease processes.


Assuntos
Citocinas/imunologia , Diabetes Mellitus Tipo 1/imunologia , Diabetes Mellitus Tipo 2/imunologia , Imunidade Inata/imunologia , Inflamassomos/imunologia , Modelos Imunológicos , Animais , Humanos , Pâncreas/imunologia
20.
J Diabetes Investig ; 15(6): 656-668, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38470018

RESUMO

Elucidating the molecular mechanism of autophagy was a landmark in understanding not only the physiology of cells and tissues, but also the pathogenesis of diverse diseases, including diabetes and metabolic disorders. Autophagy of pancreatic ß-cells plays a pivotal role in the maintenance of the mass, structure and function of ß-cells, whose dysregulation can lead to abnormal metabolic profiles or diabetes. Modulators of autophagy are being developed to improve metabolic profile and ß-cell function through the removal of harmful materials and rejuvenation of organelles, such as mitochondria and endoplasmic reticulum. Among the known antidiabetic drugs, glucagon-like peptide-1 receptor agonists enhance the autophagic activity of ß-cells, which might contribute to the profound effects of glucagon-like peptide-1 receptor agonists on systemic metabolism. In this review, the results from studies on the role of autophagy in ß-cells and their implication in the development of diabetes are discussed. In addition to non-selective (macro)autophagy, the role and mechanisms of selective autophagy and other minor forms of autophagy that might occur in ß-cells are discussed. As ß-cell failure is the ultimate cause of diabetes and unresponsiveness to conventional therapy, modulation of ß-cell autophagy might represent a future antidiabetic treatment approach, particularly in patients who are not well managed with current antidiabetic therapy.


Assuntos
Autofagia , Diabetes Mellitus , Células Secretoras de Insulina , Humanos , Células Secretoras de Insulina/fisiologia , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patologia , Autofagia/fisiologia , Animais , Diabetes Mellitus/fisiopatologia , Diabetes Mellitus/metabolismo , Hipoglicemiantes/uso terapêutico , Hipoglicemiantes/farmacologia
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