RESUMO
Atrial fibrillation (AF) is prevalent in patients with obesity and diabetes, and such patients often exhibit cardiac steatosis. Since the role of cardiac steatosis per se in the induction of AF has not been elucidated, the present study was designed to explore the relation between cardiac steatosis and AF. Transgenic (Tg) mice with cardiac-specific overexpression of perilipin 2 (PLIN2) were housed in the laboratory for more than 12 mo before the study. Electron microscopy of the atria of PLIN2-Tg mice showed accumulation of small lipid droplets around mitochondrial chains, and five- to ninefold greater atrial triacylglycerol (TAG) content compared with wild-type (WT) mice. Electrocardiography showed significantly longer RR intervals in PLIN2-Tg mice than in WT mice. Transesophageal electrical burst pacing resulted in significantly higher prevalence of sustained (>5 min) AF (69%) in PLIN2-Tg mice than in WT mice (24%), although it was comparable in younger (4-mo-old) mice. Connexin 43 (Cx43), a gap junction protein, was localized at the intercalated disks in WT atria but was heterogeneously distributed on the lateral side of cardiomyocytes in PLIN2-Tg atria. Langendorff-perfused hearts using the optical mapping technique showed slower and heterogeneous impulse propagation in PLIN2-Tg atria compared with WT atria. Cardiac overexpression of hormone-sensitive lipase in PLIN2-Tg mice resulted in atrial TAG depletion and amelioration of AF susceptibility. The results suggest that PLIN2-induced steatosis is associated with Cx43 remodeling, impaired conduction propagation, and higher incidence of AF in aged mice. Therapies targeting cardiac steatosis could be potentially beneficial against AF in patients with obesity or diabetes.
Assuntos
Fibrilação Atrial/genética , Conexina 43/metabolismo , Átrios do Coração/metabolismo , Gotículas Lipídicas/ultraestrutura , Miócitos Cardíacos/metabolismo , Perilipina-2/genética , Animais , Fibrilação Atrial/metabolismo , Fibrilação Atrial/patologia , Técnicas de Introdução de Genes , Átrios do Coração/ultraestrutura , Preparação de Coração Isolado , Camundongos , Camundongos Transgênicos , Microscopia Eletrônica , Miócitos Cardíacos/ultraestrutura , Perilipina-2/metabolismo , Esterol Esterase/genética , Esterol Esterase/metabolismo , Triglicerídeos/metabolismo , Imagens com Corantes Sensíveis à VoltagemRESUMO
Beige adipocytes in white adipose tissue (WAT) have received considerable recognition because of their potential protective effect against obesity. Pycnogenol (PYC), extracted from French maritime pine bark, has anti-inflammatory and antioxidant properties and can improve lipid profiles. However, the effect of PYC on obesity has never been explored. In this study, we investigated the effects of PYC on obesity and WAT browning in apolipoprotein E- (ApoE-) deficient mice. The results showed that PYC treatment clearly reversed body weight and the mass of eWAT gain resulting from a high-cholesterol and high-fat diet (HCD), but no difference in food intake. The morphology results showed that the size of the adipocytes in the PYC-treated mice was obviously smaller than that in the HCD-fed mice. Next, we found that PYC upregulated the expression of genes related to lipolysis (ATGL and HSL), while it decreased the mRNA level of PLIN1. PYC significantly increased the expression of UCP1 and other genes related to beige adipogenesis. Additionally, PYC increased the expression of proteins related to the protein kinase A (PKA) signaling pathway. The findings suggested that PYC decreased obesity by promoting lipolysis and WAT browning. Thus, PYC may be a novel therapeutic target for obesity.
Assuntos
Adipócitos/efeitos dos fármacos , Tecido Adiposo Marrom/efeitos dos fármacos , Tecido Adiposo Branco/efeitos dos fármacos , Apolipoproteínas E/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Flavonoides/farmacologia , Transdução de Sinais/efeitos dos fármacos , Adipócitos/metabolismo , Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Branco/metabolismo , Animais , Apolipoproteínas E/genética , Tamanho Celular/efeitos dos fármacos , Lipólise/efeitos dos fármacos , Masculino , Camundongos , Camundongos Knockout , Extratos VegetaisRESUMO
PycnogenolR (PYC), a combination of active flavonoids derived from French maritime pine bark, is a natural antioxidant that has various pharmacological activities. Here, we investigated the beneficial effect of PYC on diet-induced hepatic steatosis. Apolipoprotein E (ApoE)-deficient male mice were administered PYC at oral doses of 30 or 100 mg·kg-1·day-1 for 2 wk in advance and were then fed a high-cholesterol and -fat diet (HCD) for 8 wk. Biochemical, immunohistochemical, and gene expression analyses were conducted to explore the effect of PYC on lipid metabolism in ApoE-deficient mice on a HCD. Short-term treatment with HCD in ApoE-deficient mice induced hepatic injuries, such as lipid metabolism disorder and hepatic histopathological changes. We found that PYC reduced body weight and the increase of serum lipids that had been caused by HCD. Supplementation of PYC significantly reduced lipid deposition in the liver, as shown by the lowered hepatic lipid content and histopathological lesions. We subsequently detected genes related to lipid metabolism and inflammatory cytokines. The study showed that PYC markedly suppressed the expression of genes related to hepatic lipogenesis, fatty acid uptake, and lipid storage while increasing the lipolytic gene, which thus reduced hepatic lipid content. Furthermore, PYC mainly reduced the expression of inflammatory cytokines and the infiltration of inflammatory cells, which were resistant to the development of hepatic steatosis. These results demonstrate that PYC protects against the occurrence and development of hepatic steatosis and may provide a new prophylactic approach for nonalcoholic fatty liver disease (NAFLD).
Assuntos
Antioxidantes/farmacologia , Dieta , Flavonoides/farmacologia , Camundongos Knockout para ApoE/fisiologia , Hepatopatia Gordurosa não Alcoólica/prevenção & controle , Extratos Vegetais/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Peso Corporal/genética , Citocinas/biossíntese , Citocinas/genética , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Lipídeos/sangue , Masculino , Camundongos , Camundongos Knockout , Camundongos Knockout para ApoE/genética , Hepatopatia Gordurosa não Alcoólica/genéticaRESUMO
OBJECTIVES: This study aimed to evaluate the utility of endoscopic ultrasonography screening for small pancreatic cancer (PC) and propose a new simple scoring system for selecting individuals who should be screened. METHODS: Risk factors or symptoms related to PC were tentatively divided into high- and low-grade risk groups based mainly on reported relative risk values. Numbers of risk factors were designated as risk scores. Endoscopic ultrasonography screening was performed for 632 individuals. We analyzed scores for PC detection prospectively, and risk factors and scores of PC patients retrospectively. RESULTS: We detected 10 small malignant pancreatic neoplasms (size ≤20 mm; 8 PCs; 9 Tis or T1) and 14 advanced PCs. All small PCs and 95.5% of PCs were found in individuals with low-grade risk scores of at least 3 points (P) or high-grade risk scores of at least 1P. Both average risk scores were significantly higher in patients with small PCs (P ≤ 0.04). Cutoffs for low- and high-grade risk scores implying the presence of small PC and all PC were 3P and 1P, respectively. When subjects having one or both cutoff scores were screened, sensitivity and specificity were 100% and 64.4% for small PCs and 95.5% and 64.4% for all PCs. CONCLUSION: Endoscopic ultrasonography screening combining new scoring is effective for detecting small PC.
Assuntos
Detecção Precoce de Câncer/métodos , Endossonografia/métodos , Endossonografia/estatística & dados numéricos , Neoplasias Pancreáticas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pâncreas/patologia , Estudos Retrospectivos , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
Pancreatic neuroendocrine tumors (pNETs) are particularly rare. The various forms of PNETs, such as cystic degeneration, make differentiation from other similar pancreatic lesions difficult. We can detect small lesions by endoscopic ultrasound (EUS) and obtain preoperative pathological diagnosis by EUS-guided fine needle aspiration (FNA). We describe, here, an interesting case of pNET in a 42-year-old woman with no family history. Computed tomography and magnetic resonance imaging revealed an 18 mm × 17 mm cystic lesion with a nodule in the pancreatic tail. Two microtumors about 7 mm in diameter in the pancreatic body detected only by EUS, cystic rim and nodules all showed similar enhancement on contrast-harmonic EUS. Preoperative EUS-FNA of the microtumor was performed, diagnosing multiple pNETs. Macroscopic examination of the resected pancreatic body and tail showed that the cystic lesion had morphologically changed to a 13-mm main nodule, and 11 new microtumors (diameter 1-3 mm). Microscopically, all microtumors represented pNETs. From the findings of a broken peripheral rim on the main lesion with fibrosis, rupture of the cystic pNET was suspected. Postoperatively, pituitary adenoma and parathyroid adenoma were detected. The final diagnosis was multiple grade 1 pNETs with multiple endocrine neoplasia type 1. To the best of our knowledge, no case of spontaneous rupture of a cystic pNET has previously been reported in the English literature. Therefore, this case of very rare pNET with various morphological changes is reported.
Assuntos
Neoplasia Endócrina Múltipla Tipo 1/diagnóstico , Tumores Neuroendócrinos/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Neoplasias das Paratireoides/diagnóstico , Neoplasias Hipofisárias/diagnóstico , Adulto , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico , Feminino , Humanos , Imageamento por Ressonância Magnética , Neoplasia Endócrina Múltipla Tipo 1/patologia , Neoplasia Endócrina Múltipla Tipo 1/cirurgia , Tumores Neuroendócrinos/patologia , Tumores Neuroendócrinos/cirurgia , Pâncreas/diagnóstico por imagem , Pâncreas/patologia , Pâncreas/cirurgia , Neoplasias Pancreáticas/patologia , Neoplasias Pancreáticas/cirurgia , Neoplasias das Paratireoides/patologia , Neoplasias das Paratireoides/cirurgia , Neoplasias Hipofisárias/patologia , Neoplasias Hipofisárias/cirurgia , Estudos Retrospectivos , Ruptura Espontânea/diagnóstico , Ruptura Espontânea/patologia , Ruptura Espontânea/cirurgia , Tomografia Computadorizada por Raios XRESUMO
Cardiac intracellular lipid accumulation (steatosis) is a pathophysiological phenomenon observed in starvation and diabetes mellitus. Perilipin 2 (PLIN2) is a lipid droplet (LD)-associated protein expressed in nonadipose tissues, including the heart. To explore the pathophysiological function of myocardial PLIN2, we generated transgenic (Tg) mice by cardiac-specific overexpression of PLIN2. Tg hearts showed accumulation of numerous small LDs associated with mitochondrial chains and high cardiac triacylglycerol (TAG) content [8-fold greater than wild-type (WT) mice]. Despite massive steatosis, cardiac uptake of glucose, fatty acids and VLDL, systolic function, and expression of metabolic genes were comparable in the two genotypes, and no morphological changes were observed by electron microscopy in the Tg hearts. Twenty-four hours of fasting markedly reduced steatosis in Tg hearts, whereas WT mice showed accumulation of LDs. Although activity of adipose triglyceride lipase in heart homogenate was comparable between WT and Tg mice, activity of hormone-sensitive lipase (HSL) was 40-50% less in Tg than WT mice under both feeding and fasting conditions, suggesting interference of PLIN2 with HSL. Mice generated through crossing of PLIN2-Tg mice and HSL-Tg mice showed cardiac-specific HSL overexpression and complete lack of steatosis. The results suggest that cardiac PLIN2 plays an important pathophysiological role in the development of dynamic steatosis and that the latter was prevented by upregulation of intracellular lipases, including HSL.
Assuntos
Cardiopatias/genética , Transtornos do Metabolismo dos Lipídeos/genética , Miocárdio/metabolismo , Perilipina-2/genética , Esterol Esterase/genética , Animais , Feminino , Expressão Gênica/fisiologia , Terapia Genética/métodos , Cardiopatias/metabolismo , Cardiopatias/patologia , Cardiopatias/prevenção & controle , Transtornos do Metabolismo dos Lipídeos/metabolismo , Transtornos do Metabolismo dos Lipídeos/patologia , Transtornos do Metabolismo dos Lipídeos/prevenção & controle , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Camundongos Transgênicos , Miocárdio/patologia , Especificidade de Órgãos/genética , Perilipina-2/metabolismo , Esterol Esterase/fisiologiaRESUMO
Pycnogenol (PYC) is an extract from French maritime pine bark. Its antioxidative and anti-inflammatory effects have been shown to be beneficial for atherosclerosis. Here, we tested whether PYC could suppress high cholesterol and fat diet (HCD)-induced atherosclerosis formation in apolipoprotein E (apoE)-deficient mice. In our study, PYC suppressed oxidized low-density lipoprotein (ox-LDL)-induced lipid accumulation in peritoneal macrophages. Apolipoprotein E-deficient mice were orally administered PYC or a control solvent for ten weeks, and these mice were fed a standard diet or high cholesterol and fat diet during the latter eight weeks. Pycnogenol markedly decreased the size of atherosclerotic lesions induced by high cholesterol and fat diet compared with the nontreated controls. In addition, TLR4 expression in aortic sinus was stimulated by high cholesterol and fat diet feeding and was significantly reduced by PYC. A mechanistic analysis indicated that lipopolysaccharide (LPS) significantly increased expression of fatty acid binding protein (aP2) and macrophage scavenger receptor class A (SR-A), which were blocked by a JNK inhibitor. Furthermore, PYC inhibited the lipopolysaccharide-induced upregulation of aP2 and scavenger receptor class A via the JNK pathway. In conclusion, PYC administration effectively attenuates atherosclerosis through the TLR4-JNK pathway. Our results suggest that PYC could be a potential prophylaxis or treatment for atherosclerosis in humans.
Assuntos
Apolipoproteínas E/deficiência , Aterosclerose/metabolismo , Flavonoides/uso terapêutico , Inibidores da Agregação Plaquetária/uso terapêutico , Transdução de Sinais/fisiologia , Receptor 4 Toll-Like/biossíntese , Animais , Aterosclerose/etiologia , Aterosclerose/prevenção & controle , Colesterol na Dieta/administração & dosagem , Colesterol na Dieta/efeitos adversos , Dieta Hiperlipídica/efeitos adversos , Flavonoides/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Extratos Vegetais , Inibidores da Agregação Plaquetária/farmacologia , Transdução de Sinais/efeitos dos fármacos , Receptor 4 Toll-Like/antagonistas & inibidoresRESUMO
Over activation of microglia results in the production of proinflammatory agents that have been implicated in various brain diseases. Pycnogenol is a patented extract from French maritime pine bark (Pinus pinaster Aiton) with strong antioxidant and anti-inflammatory potency. The present study investigated whether pycnogenol may be associated with the production of proinflammatory mediators in lipopolysaccharide-stimulated BV2 (mouse-derived) microglia. It was found that pycnogenol treatment was dose-dependently associated with significantly less release of nitricoxide (NO), TNF-α, IL-6 and IL-1ß, and lower levels of intercellular adhesion molecule1 (ICAM-1) and perilipin 2 (PLIN2). Furthermore, this effect was replicated in primary brain microglia. Levels of inducible NO synthase mRNA and protein were attenuated, whereas there was no change in the production of the anti-inflammatory cytokine IL-10. Further evidence indicated that pycnogenol treatment led to the suppression of NF-κB activation through inhibition of p65 translocation into the nucleus and inhibited DNA binding of AP-1, suggesting that these proinflammatory factors are associated with NF-κB and AP-1. We conclude that pycnogenol exerts anti-inflammatory effects through inhibition of the NF-κB and AP-1pathway, and may be useful as a therapeutic agent in the prevention of diseases caused by over activation of microglia.
Assuntos
Citocinas/metabolismo , Flavonoides/farmacologia , Proteínas de Membrana/metabolismo , Microglia/efeitos dos fármacos , NF-kappa B/antagonistas & inibidores , Fator de Transcrição AP-1/metabolismo , Animais , Células Cultivadas , Relação Dose-Resposta a Droga , Flavonoides/administração & dosagem , Inflamação/metabolismo , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Lipopolissacarídeos/farmacologia , Proteínas de Membrana/genética , Camundongos , Microglia/metabolismo , NF-kappa B/genética , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Perilipina-2 , Extratos VegetaisRESUMO
OBJECTIVE: Perilipin (PLIN) 3, an intracellular lipid droplet (LD)-associated protein, is implicated in foam cell formation. Since metabolic derangements found in metabolic syndrome, such as high serum levels of glucose, insulin and free fatty acids (FFAs), are major risk factors promoting atherosclerosis, we investigated whether PLIN3 expression is affected by glucose, insulin and oleic acid (OA) using RAW264.7 cells. METHODS: Real-time PCR and Western blotting were performed to detect PLIN3 or PLIN2 expression. Oil-red O staining and Lipid Analysis were employed to measure cellular content of triacylglycerides (TAG) and cholesterol. RESULTS: PLIN3 mRNA was stimulated by high glucose or insulin concentrations individually, but not by OA. A combination of any two factors did not enhance PLIN3 expression any more than that evoked by glucose alone at 24h. Interestingly, however, simultaneous addition of all three factors synergistically enhanced the PLIN3 expression. This synergistic effect was not apparent for PLIN2 mRNA expression. Inhibitors of Src family tyrosine kinase and/or phosphatidylinositol 3-kinase, both of which are activated by insulin and FFA signaling, partially suppressed PLIN3 expression induced by the combination of the three factors. While simultaneous addition of glucose, insulin and OA remarkably increased the cellular content of TAG and cholesterol, knocking-down of PLIN3 predominantly reduced TAG content. CONCLUSIONS: These results indicate that PLIN3 expression is synergistically stimulated by high glucose, insulin and FFA concentrations, in parallel with TAG accumulation in macrophages. This finding raises new evidence of PLIN3 involvement in conversion of macrophages into foam cells.
Assuntos
Proteínas de Transporte/biossíntese , Ácidos Graxos não Esterificados/farmacologia , Glucose/farmacologia , Hipoglicemiantes/farmacologia , Insulina/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Macrófagos/metabolismo , Animais , Compostos Azo , Western Blotting , Proteínas de Transporte/genética , Colesterol/sangue , Corantes , Macrófagos/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos C57BL , Ácido Oleico/farmacologia , Perilipina-3 , Fosfatidilinositol 3-Quinases/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Transfecção , Triglicerídeos/sangue , Quinases da Família src/metabolismoRESUMO
Hemophagocytic syndrome (HPS) is an unusual disorder associated with systemic lupus erythematosus (SLE). A 64-year-old woman was admitted because of fever and urticarial vasculitis. Laboratory data revealed pancytopenia and immunological abnormalities, suggesting elevated disease activity. Prednisolone monotherapy failed to improve the pancytopenia despite the amelioration of other clinical findings. Because her condition was suggestive of HPS, tacrolimus at 2-3 mg/day was added to the prednisolone regimen. Eventually, the pancytopenia improved and prednisolone could be effectively tapered. Tacrolimus could be an additional or alternative modality for treating refractory HPS.
Assuntos
Imunossupressores/uso terapêutico , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Linfo-Histiocitose Hemofagocítica/tratamento farmacológico , Tacrolimo/uso terapêutico , Medula Óssea/patologia , Relação Dose-Resposta a Droga , Feminino , Febre , Humanos , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/patologia , Linfo-Histiocitose Hemofagocítica/complicações , Linfo-Histiocitose Hemofagocítica/patologia , Pessoa de Meia-Idade , Pancitopenia/tratamento farmacológico , Pancitopenia/etiologia , Pancitopenia/patologia , Prednisolona/uso terapêutico , Indução de Remissão , Falha de Tratamento , Resultado do Tratamento , Urticária/tratamento farmacológico , Urticária/etiologia , Urticária/patologia , Vasculite/tratamento farmacológico , Vasculite/etiologia , Vasculite/patologiaRESUMO
Excessive accumulation of lipids in macrophages results in formation of foam cells and is a hallmark of atherosclerosis. The PAT family of proteins has been implicated in this process, but details of their involvement in foam cell formation have not been fully elucidated. One of dominant members of the PAT proteins, perilipin 3 (TIP47), is likely to be involved in such a regulatory mechanism. In this study, we demonstrated that the Toll-like receptor 9 (TLR9)-mediated pathway stimulates perilipin 3 expression and accumulation of lipids, especially triglycerides, in macrophages. Oligodeoxynucleotide (ODN) 1826, a ligand of TLR9, significantly enhanced perilipin 3 expression in RAW264.7 cells, and chloroquine, a TLR9 inhibitor, almost completely inhibited ODN1826-induced perilipin 3 expression. The inhibitors of c-jun NH2-terminal kinase and PI 3-kinase suppressed the level of perilipin 3 mRNA induced by ODN1826. ODN1826 induced the expression of IL-1α and IFNß, both of which increased perilipin 3 expression. Antibodies against these cytokines suppressed the ODN1826-induced perilipin 3 mRNA levels. These results suggest that the expression of perilipin 3 in macrophages is in part regulated through the TLR9-mediated mechanism. Furthermore, ODN1826 increased intracellular lipid accumulation in the presence of oxLDL, which was reduced by perilipin 3 siRNA. Perilipin 3 expression was not stimulated by oxLDL. Depletion of perilipin 3 by siRNA specifically reduced triglyceride content in the cells but not cholesterol content, indicating that perilipin 3 is involved mainly in triglyceride accumulation. In conclusion, the TLR9-mediated pathway facilitates foam cell formation in part through increased expression of perilipin 3.
Assuntos
Aterosclerose/metabolismo , Proteínas de Transporte/metabolismo , Macrófagos/metabolismo , Receptor Toll-Like 9/metabolismo , Triglicerídeos/metabolismo , Animais , Western Blotting , Proteínas de Transporte/genética , Linhagem Celular , Cloroquina/farmacologia , Feminino , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Macrófagos/enzimologia , Camundongos , Camundongos Endogâmicos C57BL , Oligodesoxirribonucleotídeos/farmacologia , Perilipina-3 , RNA/química , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Fatty acids stimulate lipid accumulation in parallel with increased expression of adipose differentiation-related protein (ADRP) in liver cells. Although it is generally considered that the fatty acid effect on ADRP expression is mediated by peroxisome proliferator-activated receptors (PPARs), we identified here an additional molecular mechanism using the NMuLi mouse liver nonparenchymal cell line, which expresses PPARgamma and delta but not alpha. Oleic acid (OA) and specific ligands for PPARgamma and -delta stimulated ADRP expression as well as the -2,090-bp ADRP promoter activity which encompasses the PPAR response element (PPRE) adjacent to an Ets/activator protein (AP)-1 site. When the AP-1 site was mutated, OA failed to stimulate the activity despite the presence of the PPRE, whereas ligands for PPARgamma and -delta did stimulate it and so did a PPARalpha ligand under the coexpression of PPARalpha. DNA binding of AP-1 was stimulated by OA but not by PPAR ligands. Because we previously demonstrated that Pycnogenol (PYC), a French maritime pine bark extract, suppressed ADRP expression in macrophages partly by suppression of AP-1 activity, we tested the effect of PYC on NMuLi cells. PYC reduced the OA-induced ADRP expression along with suppression of lipid droplet formation. However, PYC neither suppressed the OA-stimulated ADRP promoter activity nor DNA binding of AP-1 but, instead, reduced the ADRP mRNA half-life. All these results indicate that the effect of OA on ADRP expression requires AP-1 as well as PPRE, and PYC suppresses the ADRP expression in part by facilitating mRNA degradation. PYC, a widely used dietary supplement, could be beneficial for the prevention of excessive lipid accumulation such as hepatic steatosis.
Assuntos
Flavonoides/farmacologia , Proteínas de Membrana/genética , Ácido Oleico/farmacologia , Receptores Ativados por Proliferador de Peroxissomo/fisiologia , Estabilidade de RNA/efeitos dos fármacos , Elementos de Resposta/fisiologia , Fator de Transcrição AP-1/fisiologia , Animais , Sequência de Bases , Linhagem Celular , Células Cultivadas , Ácidos Graxos/farmacologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Proteínas de Membrana/metabolismo , Camundongos , Dados de Sequência Molecular , Perilipina-2 , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Extratos Vegetais , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Elementos de Resposta/efeitos dos fármacos , Fator de Transcrição AP-1/metabolismo , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genéticaRESUMO
We conducted a survey on the use of dietary supplements and health foods (DS/HF) in definite rheumatoid arthritis (RA) patients treated by RA specialists. Among 296 patients (male 48, female 248), 179 patients (60.5%) had experience of DS/HF use. Prevalence of DS/HF use was significantly higher in female than in male patients (63.7% versus 43.8%). Overall, patients who have used DS/HF were significantly younger than those who have not used; it was particularly notable in female patients. The proportion of current users was significantly higher in those less than 5 years from diagnosis than those who had been diagnosed for 5 years or more. Products of herbs or algae (44.1%) and components of cartilage (40.8%) were the most popular DS/HF. Primary sources of product information were family members or friends (56.4%) and advertisements in the mass media (34.1%). Of the users, 73.7% did not disclose DS/HF use to their physicians. The users expected alleviation of the symptoms (35.2%) and improvement of health (34.6%). However, 59.2% of the users were unsure of the benefits. In conclusion, physicians should be aware of the high prevalence of DS/HF usage in patients with RA in Japan.
Assuntos
Artrite Reumatoide/dietoterapia , Artrite Reumatoide/tratamento farmacológico , Suplementos Nutricionais , Alimentos Orgânicos , Automedicação , Idoso , Feminino , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Inquéritos NutricionaisRESUMO
CONTEXT: Pendrin is an apical protein of thyroid follicular cells, responsible for the efflux of iodide into the follicular lumen via an iodide-chloride transport mechanism. It is unknown whether pendrin is recognized by autoantibodies. OBJECTIVE: Our objective was to examine the prevalence of pendrin antibodies in autoimmune thyroid diseases and compare with that of thyroglobulin, thyroperoxidase, TSH receptor, and sodium iodide symporter antibodies. DESIGN: In a prevalent case-control study, we analyzed the sera of 140 autoimmune thyroid disease cases (100 with Graves' disease and 40 with Hashimoto's thyroiditis) and 80 controls (50 healthy subjects, 10 patients with papillary thyroid cancer, 10 with systemic lupus erythematosus, and 10 with rheumatoid arthritis). Pendrin antibodies were measured by immunoblotting using extract of COS-7 cells transfected with pendrin and a rabbit polyclonal pendrin antibody. RESULTS: Pendrin antibodies were found in 81% of the cases and 9% of controls (odds ratio = 44; P < 0.0001). Among cases, pendrin antibodies were more frequent and of higher titers in Hashimoto's thyroiditis than in Graves' disease. Pendrin antibodies correlated significantly with thyroglobulin, thyroperoxidase, and sodium iodide symporter antibodies but not with TSH receptor antibodies. Pendrin antibodies were equally effective as thyroglobulin and thyroperoxidase antibodies in diagnosis of autoimmune thyroid diseases, especially Hashimoto's thyroiditis. CONCLUSIONS: The study identifies pendrin as a novel autoantigen recognized by patients with autoimmune thyroid diseases and proposes the use of pendrin antibodies as an accurate diagnostic tool.
Assuntos
Autoanticorpos/sangue , Autoantígenos/imunologia , Proteínas de Membrana Transportadoras/imunologia , Tireoidite Autoimune/sangue , Tireoidite Autoimune/imunologia , Animais , Reações Antígeno-Anticorpo , Autoantígenos/sangue , Autoantígenos/isolamento & purificação , Células COS , Estudos de Casos e Controles , Chlorocebus aethiops , Humanos , Iodeto Peroxidase/imunologia , Proteínas de Ligação ao Ferro/imunologia , Proteínas de Membrana Transportadoras/genética , Sensibilidade e Especificidade , Estudos Soroepidemiológicos , Transportadores de Sulfato , Simportadores/imunologia , Tireoglobulina/imunologia , Tireoidite Autoimune/diagnósticoRESUMO
Mycobacterium leprae survives and replicates within a lipid droplet stored in the enlarged phagosome of histiocytes, a typical feature of lepromatous leprosy that is thought to be an important nutrient source for the bacillus. However, the underlying mechanisms by which lipids accumulate within phagosomes remain unclear. Recently, it was revealed that the lipid droplet-associated proteins, including ADRP and perilipin, play essential roles in lipid accumulation in adipocytes or macrophages. Therefore, we attempted to examine the role of these proteins in leprosy pathogenesis. ADRP and perilipin localized to the phagosomal membrane, which contains M. leprae in skin biopsy specimens of lepromatous leprosy. ADRP expression was transiently increased after phagocytosis in THP-1 cells. However, high levels of ADRP expression persisted only when live M. leprae, but not dead bacilli or latex beads, was added. Furthermore, although peptidoglycan, a Toll-like receptor 2 ligand, suppressed the expression levels of ADRP and perilipin, M. leprae infection inhibited this suppression. These results suggest that live M. leprae has the ability to actively induce and support ADRP/perilipin expression to facilitate the accumulation of lipids within the phagosome and to further maintain a suitable environment for the intracellular survival within the macrophage.
Assuntos
Regulação da Expressão Gênica , Hanseníase Virchowiana/metabolismo , Macrófagos/microbiologia , Proteínas de Membrana/metabolismo , Mycobacterium leprae/patogenicidade , Fosfoproteínas/metabolismo , Animais , Proteínas de Transporte , Linhagem Celular , Humanos , Hanseníase Virchowiana/patologia , Macrófagos/metabolismo , Proteínas de Membrana/genética , Camundongos , Camundongos Nus , Perilipina-1 , Perilipina-2 , Fosfoproteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Pele/metabolismo , Pele/microbiologia , Pele/patologiaRESUMO
Adipose differentiation-related protein (ADRP) is highly expressed in macrophages and human atherosclerotic lesions. We demonstrated that Toll-like receptor (TLR) 4-mediated signals, which are involved in atherosclerosis formation, enhanced the expression of ADRP in macrophages. Lipopolysaccharide (LPS) enhanced the ADRP expression in RAW264.7 cells or peritoneal macrophages from wild-type mice, but not in macrophages from TLR4-deficient mice. Actinomycin D almost completely abolished the LPS effect, whereas cycloheximide decreased the expression at 12 h, indicating that the LPS-induced ADRP expression was stimulated at the transcriptional level and was also mediated by new protein synthesis. LPS enhanced the ADRP promoter activity, in part, by stimulating activator protein (AP)-1 binding to the Ets/AP-1 element. In addition, preceding the increase of the ADRP mRNA, LPS induced the expression of interleukin (IL)-6, IL-1alpha, and interferon-beta mRNAs, all of which stimulated the ADRP expression. Antibodies against these cytokines or inhibitors of c-Jun NH(2)-terminal kinase and nuclear factor (NF)-kappaB suppressed the ADRP mRNA level. Thus TLR4 signals stimulate the ADRP expression both in direct and indirect manners. Pycnogenol (PYC), an extract of French maritime pine, suppressed the expression of ADRP and the above-mentioned cytokines. PYC suppressed the ADRP promoter activity and enhancer activity of AP-1 and NF-kappaB, whereas it did not affect the LPS-induced DNA binding of these factors. In conclusion, TLR4-mediated signals stimulate the ADRP expression in macrophages while PYC antagonizes this process. PYC, a widely used dietary supplement, might be useful for prevention of atherosclerosis.
Assuntos
Flavonoides/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Proteínas de Membrana/genética , Receptor 4 Toll-Like/antagonistas & inibidores , Animais , Células Cultivadas , Lipopolissacarídeos/farmacologia , Macrófagos/metabolismo , Proteínas de Membrana/metabolismo , Camundongos , Perilipina-2 , Pinus/química , Extratos Vegetais/farmacologia , RNA Mensageiro/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Receptor 4 Toll-Like/metabolismo , Receptor 4 Toll-Like/fisiologiaAssuntos
Tireoidite Supurativa , Antibacterianos/administração & dosagem , Diagnóstico Diferencial , Infecções por Escherichia coli , Humanos , Infecções por Pseudomonas , Infecções Estafilocócicas , Infecções Estreptocócicas , Tireoidite Supurativa/diagnóstico , Tireoidite Supurativa/microbiologia , Tireoidite Supurativa/fisiopatologia , Tireoidite Supurativa/terapiaRESUMO
ADRP is associated with intracellular lipid droplets. We demonstrate the regulatory mechanism for ADRP expression in RAW264.7 macrophages. The ADRP mRNA expression was stimulated by PMA, and synergistically enhanced in association with its protein level in the presence of lipids. A proteasome inhibitor protected the protein from degradation under the lipid-free conditions. One of the possible sites of the PMA action was proved to be an Ets/AP-1 element in the promoter, since mutations of this site reduced the PMA-induced promoter activity, and ligation of this element led to a significant increase in the PMA-responsiveness of homologous or heterologous promoters. Mutations of this site diminished the synergistic effect on the promoter activity induced by PMA and oleic acid, suggesting a possible interaction between this site and the downstream PPARdelta site. EMSA revealed that PU.1 and AP-1 conjointly bound to this site. The juxtaposition of the two sequences was requisite for full activity, since spacer sequences between them decreased the PMA-induced activity. PI3 kinase inhibitor was found to reduce the PMA-induced mRNA expression and promoter activity in parallel with PU.1/AP-1 complex formation on EMSA. From these results, we concluded that the Ets/AP-1 site is an important cis-acting element that regulates the ADRP gene expression in macrophages.
Assuntos
Regulação da Expressão Gênica , Macrófagos/metabolismo , Proteínas de Membrana/biossíntese , Proteínas Proto-Oncogênicas/metabolismo , Fator de Transcrição Sp1/genética , Transativadores/metabolismo , Fator de Transcrição AP-1/metabolismo , Tecido Adiposo , Animais , Células Cultivadas , Expressão Gênica/efeitos dos fármacos , Proteínas de Membrana/genética , Camundongos , Dados de Sequência Molecular , PPAR delta , Perilipina-2 , Regiões Promotoras Genéticas , RNA Mensageiro/biossíntese , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
A novel human tumor-associated antigen, receptor-binding cancer antigen expressed on SiSo cells (RCAS1), induces apoptosis in normal human erythroid progenitor cells, which express putative RCAS1 receptors. In the present study, we investigated a possible role of RCAS1 produced by human peripheral blood monocytes (CD14-positive cells) and monocyte-derived macrophages. RCAS1 was immunohistochemically detected in monocytes as well as macrophages. When macrophages were stimulated with lipopolysaccharide (LPS), the expression of RCAS1 was remarkably enhanced. An increased production of RCAS1 mRNA was observed in LPS-stimulated macrophages by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Soluble RCAS1 molecules were only detected in the culture supernatants obtained from LPS-stimulated macrophages. Moreover, LPS-stimulated macrophages induced cell death of erythroid progenitor cells through RCAS1 production. These results suggest that macrophages may negatively regulate erythropoiesis at least in part through the production of RCAS1 molecules, and this may contribute to the pathogenesis of the anemia seen in patients with inflammatory disorders.