Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.929
Filtrar
1.
Life Sci ; 240: 117019, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31678554

RESUMO

AIMS: Long noncoding RNA melanotransferrin antisense RNA (MFI2-AS1) plays a vital role in the development of multiple diseases. This study aimed to investigate the effect of this lncRNA on osteoarthritis progression and explore the interaction among MFI2-AS1, microRNA (miR)-130a-3p and transcription factor 4 (TCF4). METHODS: Forty-six knee osteoarthritis tissues and 28 normal samples were collected. Human chondrocytes C28/I2 cells treated by lipopolysaccharide (LPS) were used as the model of osteoarthritis. The expression levels of MFI2-AS1, miR-130a-3p and TCF4 were detected by quantitative real-time polymerase chain reaction or western blot. LPS-induced chondrocytes injury was investigated by cell viability, apoptosis, inflammatory response and extracellular matrix degradation using MTT, flow cytometry, enzyme-linked immunosorbent assay and western blot. The target association between miR-130a-3p and MFI2-AS1 or TCF4 was confirmed by luciferase reporter assay and RNA immunoprecipitation. RESULTS: MFI2-AS1 expression was increased in osteoarthritis tissues and LPS-treated C28/I2 cells. Silence of MFI2-AS1 attenuated LPS-induced viability suppression, apoptosis production, inflammatory response and extracellular matrix degradation. MFI2-AS1 was validated as a decoy of miR-130a-3p and TCF4 was confirmed as a target of miR-130a-3p. miR-130a-3p overexpression inhibited LPS-induced cell injury in C28/I2 cells by decreasing TCF4 expression. Moreover, knockdown of MFI2-AS1 alleviated LPS-induced cell injury in C28/I2 cells by mediating miR-130a-3p and TCF4. CONCLUSION: Knockdown of MFI2-AS1 increased cell viability but suppressed apoptosis, inflammatory response and extracellular matrix degradation in LPS-treated chondrocytes by increasing miR-130a-3p and decreasing TCF4, indicating a novel target for the treatment of osteoarthritis.


Assuntos
Técnicas de Silenciamento de Genes , Lipopolissacarídeos , MicroRNAs/genética , Osteoartrite/genética , Osteoartrite/prevenção & controle , RNA Antissenso/genética , RNA Longo não Codificante/genética , Fator de Transcrição 4/genética , Animais , Linhagem Celular , Proliferação de Células , Sobrevivência Celular , Condrócitos , Humanos
2.
Toxicol Lett ; 321: 122-130, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31874197

RESUMO

Our previous studies confirmed that prenatal caffeine exposure (PCE) could induce susceptibility to osteoarthritis in adult offspring rats due to poor chondrocyte differentiation, but its mechanism remains to be further investigated. This study aimed to explore whether subchondral bone dysplasia mediates susceptibility to osteoarthritis in adult offspring rats induced by PCE. Pregnant Wistar rats were treated with caffeine (120 mg/kg.d) or saline from gestational day (GD) 9 to 20. The female offspring were euthanized to collect femurs at GD20, postnatal week (PW) 6, and PW28 (non-ovariectomy and ovariectomy groups) to detect osteoarthritis-like phenotype, subchondral bone mass, ossification center development, and other evidence. The results showed that PCE increased the Mankin score of pathological articular cartilage, but decreased articular cartilage thickness and subchondral bone mass, which were more obvious after ovariectomy. Meanwhile, the correlation analysis results demonstrated that the Mankin score of articular cartilage was significantly negatively correlated with subchondral bone mass, and the thickness of articular cartilage was significantly positively correlated with subchondral bone mass. Further, the length and area of the primary and secondary ossification centers, the number of osteoblasts, and the related genes' expression of osteogenic differentiation (e.g., Runx2, BSP, ALP, and OCN) were all significantly decreased in the PCE group before and after birth. Taken together, PCE induced susceptibility to osteoarthritis in adult female offspring, which was likely related to the subchondral bone dysplasia and reduction of subchondral bone mass production due to developmental disorder of primary and secondary ossification centers caused by osteoblast differentiation disability before and after birth.


Assuntos
Doenças do Desenvolvimento Ósseo/induzido quimicamente , Cafeína/toxicidade , Cartilagem Articular/efeitos dos fármacos , Estimulantes do Sistema Nervoso Central/toxicidade , Osteoartrite/induzido quimicamente , Osteogênese/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal , Fatores Etários , Animais , Doenças do Desenvolvimento Ósseo/genética , Doenças do Desenvolvimento Ósseo/metabolismo , Doenças do Desenvolvimento Ósseo/patologia , Cartilagem Articular/patologia , Diferenciação Celular/efeitos dos fármacos , Feminino , Fêmur/efeitos dos fármacos , Fêmur/metabolismo , Fêmur/patologia , Idade Gestacional , Osteoartrite/genética , Osteoartrite/metabolismo , Osteoartrite/patologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoblastos/patologia , Osteogênese/genética , Ovariectomia , Gravidez , Ratos Wistar , Fatores Sexuais , Tíbia/efeitos dos fármacos , Tíbia/metabolismo , Tíbia/patologia
3.
Pediatrics ; 144(6)2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31722963

RESUMO

OBJECTIVES: In this study, we aimed to contrast the bacteriologic epidemiology of osteoarticular infections (OAIs) between 2 patient groups in successive 10-year periods, before and after the extensive use of nucleic acid amplification assays in the diagnostic process. METHODS: Epidemiologic data and bacteriologic etiologies of all children presenting with OAIs on admission to our institution over 20 years (1997-2016) were assessed retrospectively. The population was divided into 2 cohorts, using the standardized use of polymerase chain reaction as the cutoff point (2007). The conventional cohort included children with OAIs mainly investigated by using classic cultures, whereas the molecular cohort referred to patients also investigated by using molecular assays. RESULTS: Kingella kingae was the most frequently isolated pathogen, responsible for 51% of OAIs, whereas other classic pathogens were responsible for 39.7% of cases in the molecular cohort. A statistically significant increase in the mean incidence of OAIs was observed, as was a decrease in the mean age at diagnosis after 2007. After 2007, the pathogen remained unidentified in 21.6% of OAIs in our pediatric population. CONCLUSIONS: Extensive use of nucleic acid amplification assays improved the detection of fastidious pathogens and has increased the observed incidence of OAI, especially in children aged between 6 and 48 months. We propose the incorporation of polymerase chain reaction assays into modern diagnostic algorithms for OAIs to better identify the bacteriologic etiology of OAIs.


Assuntos
Artrite Infecciosa/diagnóstico , Kingella kingae/isolamento & purificação , Infecções por Neisseriaceae/diagnóstico , Osteoartrite/diagnóstico , Osteomielite/diagnóstico , Adolescente , Artrite Infecciosa/genética , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Humanos , Lactente , Recém-Nascido , Kingella kingae/genética , Masculino , Infecções por Neisseriaceae/genética , Osteoartrite/genética , Osteomielite/genética , Reação em Cadeia da Polimerase em Tempo Real/métodos , Estudos Retrospectivos
4.
Yonsei Med J ; 60(11): 1081-1092, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31637891

RESUMO

PURPOSE: Accumulating evidence suggests that microRNA-145 (miR-145) plays an important role in osteoarthritis (OA), which is a chronic progressive joint disease. Long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) promotes metastasis in cancers and functions as a sponge for miR-145. However, the role of MALAT1/miR-145 in OA pathogenesis has not yet been elucidated. MATERIALS AND METHODS: The expression of MALAT1 and miR-145 was examined by quantitative real-time PCR; the interaction between miR-145, MALAT1 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) 5 was verified by luciferase reporter assay. Correlations among MALAT1, miR-145, and ADAMTS5 were analyzed by Spearman rank analysis. Chondrocytes viability and cartilage extracellular matrix (ECM) degradation were investigated with cell viability assay and Western blotting analyzing expression of ADAMTS5, collagen type 2 alpha 1 (COL2A1), aggrecan (ACAN), and cartilage oligomeric matrix protein (COMP). RESULTS: MALAT1 was upregulated, and miR-145 was downregulated in OA samples and IL-1ß-induced chondrocytes. Mechanically, miR-145 could directly bind to MALAT1 and ADAMTS5. Moreover, miR-145 expression was negatively correlated with MALAT1 and ADAMTS5 expression in OA patients, whereas MALAT1 and ADAMTS5 expression was positively correlated. Functionally, overexpression of MALAT1 inhibited chondrocyte viability and promoted cartilage ECM degradation in IL-1ß-induced chondrocytes. In support thereof, MALAT1 silencing and miR-145 upregulation exerted the opposite effect in IL-1ß-induced chondrocytes. Moreover, the effect of MALAT1 was counteracted by miR-145 upregulation, and ADAMTS5 restoration could abate miR-145 effects. CONCLUSION: An MALAT1/miR-145 axis contributes to ECM degradation in IL-1ß-induced chondrocytes through targeting ADAMTS5, suggesting that MALAT1/miR-145/ADAMTS5 signaling may underlie human OA pathogenesis.


Assuntos
Proteína ADAMTS5/genética , Cartilagem Articular/patologia , Condrócitos/patologia , Matriz Extracelular/metabolismo , Interleucina-1beta/efeitos adversos , MicroRNAs/metabolismo , Osteoartrite/genética , RNA Longo não Codificante/metabolismo , Proteína ADAMTS5/metabolismo , Adulto , Idoso , Sequência de Bases , Sobrevivência Celular/genética , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , MicroRNAs/genética , Osteoartrite/patologia , RNA Longo não Codificante/genética
5.
Nat Commun ; 10(1): 4898, 2019 10 25.
Artigo em Inglês | MEDLINE | ID: mdl-31653858

RESUMO

Osteoarthritis (OA) is a prevalent degenerative disease, which involves progressive and irreversible destruction of cartilage matrix. Despite efforts to reconstruct cartilage matrix in osteoarthritic joints, it has been a difficult task as adult cartilage exhibits marginal repair capacity. Here we report the identification of tankyrase as a regulator of the cartilage anabolism axis based on systems-level factor analysis of mouse reference populations. Tankyrase inhibition drives the expression of a cartilage-signature matrisome and elicits a transcriptomic pattern that is inversely correlated with OA progression. Furthermore, tankyrase inhibitors ameliorate surgically induced OA in mice, and stem cell transplantation coupled with tankyrase knockdown results in superior regeneration of cartilage lesions. Mechanistically, the pro-regenerative features of tankyrase inhibition are mainly triggered by uncoupling SOX9 from a poly(ADP-ribosyl)ation (PARylation)-dependent protein degradation pathway. Our findings provide insights into the development of future OA therapies aimed at reconstruction of articular cartilage.


Assuntos
Cartilagem Articular/efeitos dos fármacos , Condrócitos/metabolismo , Matriz Extracelular/efeitos dos fármacos , Transplante de Células-Tronco Mesenquimais , Osteoartrite do Joelho/metabolismo , Poli ADP Ribosilação/efeitos dos fármacos , Fatores de Transcrição SOX9/efeitos dos fármacos , Tanquirases/antagonistas & inibidores , Animais , Cartilagem Articular/metabolismo , Cartilagem Articular/fisiologia , Simulação por Computador , Inibidores Enzimáticos , Matriz Extracelular/metabolismo , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Camundongos , Osteoartrite/genética , Osteoartrite/metabolismo , Osteoartrite do Joelho/genética , Poli ADP Ribosilação/fisiologia , Ratos , Regeneração/genética , Fatores de Transcrição SOX9/metabolismo , Tanquirases/genética , Tanquirases/metabolismo
6.
BMC Complement Altern Med ; 19(1): 264, 2019 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-31590658

RESUMO

BACKGROUND: Osteoarthritis (OA) is a common degenerative disease of synovial joints caused by inflammation. Acteoside (ACT), a major component and lipase inhibitor from the Chinese tea Ligustrum purpurascens kudingcha, has been reported to regulate the inflammation and immune response. The study aims to investigate the effects of ACT on inflammatory responses and joint protection in OA rats. METHODS: Cell proliferation was examined by MTT and colony formation assay. Apoptosis was analyzed using flow cytometry with Annexin V/PI staining. ELISA was employed to examine the concentration of inflammatory cytokines. OA rat model was established by surgery stimulation. RESULTS: ACT treatment significantly inhibited the upregulation of inflammatory cytokines induced by IL-1ß in primary chondrocytes, including IL-6, IL-12, TNF-α and IFN-γ. ACT stimulation also enhanced the cell proliferation, while inhibited cell apoptosis in IL-1ß-treated chondrocytes. Consistently, ACT treatment led to downregulation of cleaved-caspase-3 and apoptosis regulator Bax, and upregulation of Bcl-2. Furthermore, ACT treatment inhibited IL-1ß-induced activation of JAK/STAT pathway. The results were confirmed in surgery-induced OA rat model. Moreover, ACT treatment significantly inhibited synovial inflammation and articular chondrocyte apoptosis in OA rats. CONCLUSION: Our findings indicate that ACT has the potential therapeutic effect on OA through inhibiting the inflammatory responses via inactivating JAK/STAT signaling pathway.


Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Glucosídeos/administração & dosagem , Ligustrum/química , Osteoartrite/tratamento farmacológico , Fenóis/administração & dosagem , Animais , Proliferação de Células/efeitos dos fármacos , Condrócitos , Modelos Animais de Doenças , Humanos , Interferon gama/genética , Interferon gama/imunologia , Janus Quinases/genética , Janus Quinases/imunologia , Masculino , Osteoartrite/genética , Osteoartrite/imunologia , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição STAT/genética , Fatores de Transcrição STAT/imunologia , Transdução de Sinais/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia
7.
Clin Exp Rheumatol ; 37 Suppl 120(5): 40-47, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31621575

RESUMO

MicroRNAs are small double-stranded RNAs, which negatively regulate gene expression and have been shown to have key roles in both chondrocyte development and cartilage homeostasis with age. Deletion of all microRNAs in chondrocytes leads to skeletal growth defects in mice, whilst deletion of specific microRNAs, e.g. miR-140, leads to premature articular cartilage degradation and increased susceptibility to posttraumatic osteoarthritis. Studies comparing microRNA expression in normal human articular cartilage compared to osteoarthritic cartilage show differential expression, but varying sample groups make interpretation difficult. MicroRNAs have been proposed as circulating biomarkers of osteoarthritis, but again, this differs amongst patient cohorts. Many micro-RNAs have been shown to have roles in chondrocyte phenotype via signalling pathways, apoptosis, autophagy and senescence. Modulating microRNAs in the joint has been shown to reduce osteoarthritis in animal models and translating this to man as a novel therapeutic strategy will be key.


Assuntos
Autofagia , Cartilagem Articular , MicroRNAs , Osteoartrite , Animais , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Condrócitos/metabolismo , Condrócitos/patologia , Humanos , Masculino , Camundongos , MicroRNAs/genética , MicroRNAs/fisiologia , Osteoartrite/genética , Osteoartrite/metabolismo
8.
J Biosci ; 44(4)2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31502578

RESUMO

Chondrosenescence (chondrocyte senescence) and subchondral bone deterioration in osteoarthritic rats were analyzed after treatment with the estrogenic herb Labisia pumila (LP) or diclofenac. Osteoarthritis (OA) was induced in bilaterally ovariectomized (OVX) rats by injecting mono-iodoacetate into the right knee joints. Rats were grouped (n = 8) into nontreated OVX+OA control, OVX+OA + diclofenac (5 mg/kg) (positive control), OVX+OA + LP leaf extract (150 and 300 mg/kg) and healthy sham control. After 8 weeks' treatment, their conditions were evaluated via serum biomarkers, knee joint histology, bone histomorphometry, protein and mRNA expressions. The LP significantly reduced cartilage erosion, femur bone surface alteration, bone loss and porosity and increased trabecular bone thickness better than diclofenac and the non-treated OA. The cartilage catabolic markers' (matrix metalloproteinase (MMP)-13, RUNX2, COL10a, ERa, CASP3 and HIF-2 alpha) mRNA expressions were down-regulated and serum bone formation marker, PINP, was increased by LP in a dose-dependent manner. The LP (containing myricetin and gallic acid) showed protection against chondrosenescence, chondrocyte death, hypoxia-induced cartilage catabolism and subchondral bone deterioration. The bone and cartilage protective effects were by suppressing proteases (collagen break-down), bone resorption and upregulating subchondral bone restoration. The cartilage ER alpha over-expression showed a strong positive correlation with MMP-13, COL10 alpha1, histological, micro-computed tomography evidence for cartilage degradation and chondrosenescence.


Assuntos
Envelhecimento/efeitos dos fármacos , Receptor alfa de Estrogênio/genética , Osteoartrite/tratamento farmacológico , Extratos Vegetais/farmacologia , Primulaceae/química , Envelhecimento/genética , Animais , Desenvolvimento Ósseo/efeitos dos fármacos , Cartilagem/efeitos dos fármacos , Cartilagem/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Diclofenaco/farmacologia , Modelos Animais de Doenças , Flavonoides/farmacologia , Ácido Gálico/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Iodoacetatos/farmacologia , Metaloproteinase 13 da Matriz/genética , Metabolismo/efeitos dos fármacos , Osteoartrite/genética , Osteoartrite/patologia , Ovariectomia , Extratos Vegetais/química , Ratos
9.
Food Funct ; 10(9): 6135-6146, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31497826

RESUMO

Osteoarthritis (OA) is a degenerative joint disease, whose progression is closely related to the inflammatory environment. Urolithin A (UA), a natural metabolite of a class of compounds (ellagitannins and ellagic acid) found in pomegranates and other fruits and nuts, has been proved to exert anti-inflammatory effects in a variety of diseases. However, the exact role of UA in OA development is still unclear. In the present study, we examined the latent mechanism of UA and its protective role in the progression of OA by both in vitro and in vivo experiments. In vitro, UA inhibited the interleukin-1 beta (IL-1ß) induced over-production of nitric oxide (NO), prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in a concentration-dependent manner in human OA chondrocytes. Furthermore, by downregulating the expression of metalloproteinase 13 (MMP13) and thrombospondin motifs 5 (ADAMTS5), UA attenuated the degradation of the extracellular matrix (ECM) induced by IL-1ß. Mechanistically, UA was found to suppress the activation of PI3K/Akt/NF-κB pathways. In vivo, in a surgically induced mouse OA model, UA-induced protective effects in OA development could be detected. In summary, this research suggested that UA may be adopted as a new therapeutic agent for the treatment of OA.


Assuntos
Cumarínicos/administração & dosagem , Interleucina-1beta/imunologia , NF-kappa B/imunologia , Osteoartrite/tratamento farmacológico , Fosfatidilinositol 3-Quinases/imunologia , Proteínas Proto-Oncogênicas c-akt/imunologia , Animais , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Dinoprostona/imunologia , Humanos , Interleucina-1beta/genética , Interleucina-6/genética , Interleucina-6/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Óxido Nítrico/imunologia , Osteoartrite/genética , Osteoartrite/imunologia , Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/efeitos dos fármacos
10.
Arch Med Res ; 50(3): 86-90, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31495394

RESUMO

BACKGROUND AND AIMS: The chemokine (C-C motif) Ligand 2 (CCL2)/CCR2 signaling was associated with macrophage accumulation, synovitis and cartilage damage in a mouse osteoarthritis (OA) model. MATERIALS AND METHODS: Here a case-control study in a Chinese Han population was conducted to investigate the possible association between the CCL2 gene polymorphism and risk of OA. DNA was extracted from 367 primary knee OA patients and 303 healthy controls. Then CCL2 gene polymorphisms were determined using a standard polymerase chain reaction restriction fragment length polymorphism. Plasma CCL2 levels were measured by using sandwich enzyme-linked immunosorbent assay (ELISA). RESULTS: It was found the CCL2 gene rs1024611 and rs4586 polymorphisms significantly increased the risk of OA. Stratified analyses showed the risk of OA might be increased by rs1024611 polymorphism in males and non-drinkers, and was increased by rs4586 polymorphism among smokers and drinkers. The CC genotype of rs4586 polymorphism was significantly correlated with the increased CCL2 level compared to TT genotype. CONCLUSIONS: In conclusion, CCL2 gene polymorphisms (rs1024611 and rs4586) confer susceptibility to OA and may be potential markers for early diagnosis of OA.


Assuntos
Quimiocina CCL2/genética , Predisposição Genética para Doença/genética , Osteoartrite/genética , Idoso , Animais , Grupo com Ancestrais do Continente Asiático/genética , Estudos de Casos e Controles , Quimiocina CCL2/metabolismo , Feminino , Genótipo , Humanos , Masculino , Camundongos , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição/genética , Polimorfismo de Nucleotídeo Único/genética
11.
Mol Med Rep ; 20(4): 3867-3873, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31485657

RESUMO

Osteoarthritis (OA) is one of the most prevalent types of chronic joint diseases. Chondrocytes survival is closely associated with the destruction of joints in patients with OA. Long noncoding RNAs (lncRNAs) serve a critical role in OA. However, to the best of our knowledge, the role of lncRNAs NR024118 in OA has not been examined. In the present study, the expression levels of NR024118 in lipopolysaccharide (LPS)­induced chondrocytes was measured using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) and the apoptosis levels of cells was determined using flow cytometry. The levels of scavenged reactive oxygen species and expression levels of the antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and heme oxygenase­1 (HO­1) were measured using specialized detection kits. The expression of interleukin (IL)­1ß, IL­6 and IL­18 were measured using ELISA. Expression of the cell apoptosis markers Bcl­2, Bax, Bcl­2­like protein 11, NF­κB, phosphorylated (p)­NF­κB inhibitor ß (IκBß), IκBß, p­transcription factor p65 (p65) and p65, and nuclear factor erythroid­2 related factor 2 (Nrf2) signaling pathways­associated proteins, Nrf2, HO­1 and quinone oxidoreductase­1 were detected by western blot analysis and RT­qPCR. The results indicated that in ATDC5 cells, apoptosis, oxidative stress and inflammation were significantly increased and the expression level of NR024118 was significantly decreased by LPS­mediated induction. NR024118 overexpression significantly reversed the effects of LPS treatment in the ATDC5 cell line. In addition, the overexpression of NR024118 decreased NF­κB expression levels and activated the Nrf2 signaling pathways in LPS­induced ATDC5 cells. The present study demonstrated that NR024118 attenuated the effects of LPS­induction on ATDC5 cells. These results suggest that NR024118 may be a potential target for diagnosis and treatment of OA.


Assuntos
Apoptose , Condrócitos/imunologia , Inflamação/genética , Fator 2 Relacionado a NF-E2/imunologia , NF-kappa B/imunologia , RNA Longo não Codificante/genética , Animais , Linhagem Celular , Condrócitos/metabolismo , Inflamação/imunologia , Lipopolissacarídeos/imunologia , Camundongos , Osteoartrite/genética , Osteoartrite/imunologia , Transdução de Sinais , Regulação para Cima
12.
Int J Med Sci ; 16(8): 1057-1071, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31523167

RESUMO

Synovitis in osteoarthritis (OA) the consequence of low grade inflammatory process caused by cartilage breakdown products that stimulated the production of pro-inflammatory mediators by fibroblast-like synoviocytes (FLS). FLS participate in joint homeostasis and low grade inflammation in the joint microenvironment triggers FLS transformation. In the current study, we aimed to identify differentially expressed genes and potential miRNA regulations in human OA FLS through deep sequencing and bioinformatics approaches. The 245 differentially expressed genes in OA FLS were identified, and pathway analysis using various bioinformatics databases indicated their enrichment in functions related to altered extracellular matrix organization, cell adhesion and cellular movement. Moreover, among the 14 dysregulated genes with potential miRNA regulations identified, src kinase associated phosphoprotein 2 (SKAP2), adaptor related protein complex 1 sigma 2 subunit (AP1S2), PHD finger protein 21A (PHF21A), lipoma preferred partner (LPP), and transcription factor AP-2 alpha (TFAP2A) showed similar expression patterns in OA FLS and OA synovial tissue datasets in Gene Expression Omnibus database. Ingenuity Pathway Analysis identified the dysregulated LPP participated in cell migration and cell spreading of OA FLS, which was potentially regulated by miR-141-3p. The current findings suggested new perspectives into understanding the novel molecular signatures of FLS involved in the pathogenesis of OA, which may be potential therapeutic targets.


Assuntos
Osteoartrite/genética , Osteoartrite/patologia , Sinoviócitos/fisiologia , Transcriptoma , Adulto , Adesão Celular/genética , Movimento Celular/genética , Células Cultivadas , Biologia Computacional , Proteínas do Citoesqueleto/genética , Bases de Dados Genéticas , Matriz Extracelular/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Proteínas com Domínio LIM/genética , MicroRNAs/genética , Família Multigênica , Mapas de Interação de Proteínas/genética , RNA Mensageiro , Reprodutibilidade dos Testes , Sinoviócitos/patologia
13.
Clin Lab ; 65(8)2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31414759

RESUMO

BACKGROUND: The current study aims to verify the abnormal expression of miR-29b-3p in peripheral blood mono-nuclear cells and synovial fluid of patients with osteoarthritis (OA), and to explore whether this abnormality is related to the severity of OA. METHODS: The level of miR-29b-3p was detected by real-time quantitative PCR in peripheral blood mononuclear cells (PBMCs) and joint fluid. Kellgren-Lawrence X-ray grading was performed in patients with OA, and the correlation between miR-29b-3p and age, weight and Kellgren-Lawrence X-ray grading was analyzed. RESULTS: The expression of miR-29b-3p in PBMCs of patients with OA was higher than that of the healthy control group ((4.48 ± 2.44) vs. (7.40 ± 4.10), p < 0.05), and the difference was statistically significant. In the synovial fluid of OA patients, the expression of miR-29b-3p was higher than that of the non-OA group ((4.90 ± 2.33) vs. (77.55 ± 2.79), p < 0.05), and the difference was statistically significant. Meanwhile, the relative expression of miR-29b-3p in PBMCs of patients with OA was positively correlated with age, body mass index (BMI), Kellgren-Lawrence X-ray classification (r = 0.896, p < 0.001; r = 0.880, p < 0.001; r = 0.357, p = 0.005). CONCLUSIONS: The high expression of miR-29b-3p in PBMCs and synovial fluid of patients with OA was positively correlated with age, body mass index and Kellgren-Lawrence X-ray classification. Therefore, miR-29b-3p may be involved in the pathogenesis of OA, and miR-29b-3p may be used as a potential biomarker for OA.


Assuntos
Biomarcadores/metabolismo , Leucócitos Mononucleares/metabolismo , MicroRNAs/genética , Osteoartrite/genética , Líquido Sinovial/metabolismo , Adulto , Idoso , Índice de Massa Corporal , Células Cultivadas , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/diagnóstico
14.
Food Funct ; 10(9): 5697-5706, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31435629

RESUMO

Osteoarthritis (OA) is a common form of arthritis, which is characterized by the degeneration of articular cartilage, leading to joint dysfunction. Oral drug therapy seems to ameliorate some signs and symptoms of OA, but may be accompanied by side effects and does not appear to be effective long-term. Seaweed has received much attention for pharmacological application due to its various biomedical properties, including the anti-inflammation, antitumor, and antioxidant effects. This study investigated the ameliorative effects of a dietary polysaccharide from Eucheuma cottonii extract (ECE) on an anterior cruciate ligament transection with partial medial meniscectomy surgery (ACLT+MMx) to induce OA in high-fat diet (HFD)-induced obese rats. Male Sprague-Dawley rats were fed an HFD for 12 weeks before ACLT+MMx surgery, after which they were administered a daily oral gavage of saline (Sham, OB Sham, and OBOA) and either low-dose ECE (100 mg per kg body weight), high-dose ECE (400 mg per kg body weight), or glucosamine sulfate as a positive control (OBOAGS; 200 mg per kg body weight) for 5 weeks. Treatment with ECE decreased the body weight, triglyceride and total cholesterol (TC) levels, and the TC/high-density lipoprotein (HDL)-C ratio in the obese rats. Additionally, ECE downregulated the expression of proinflammatory cytokines, including tumor necrosis factor-α, interleukin-1ß, and leptin, and suppressed nuclear factor-kappa B and extracellular-signal-regulated kinase-1/2 expression, resulting in a decrease in the levels of matrix metalloproteinase (MMP)-1 and MMP-13 and prostaglandin-E2 and attenuated cartilage degradation. These results demonstrate that the dietary polysaccharide from ECE can suppress OA development in obese rats, suggesting its potential efficacy as a promising candidate for OA treatment.


Assuntos
Cartilagem Articular/efeitos dos fármacos , Citocinas/imunologia , Obesidade/complicações , Osteoartrite/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Polissacarídeos/administração & dosagem , Rodófitas/química , Alga Marinha/química , Animais , Ligamento Cruzado Anterior/efeitos dos fármacos , Ligamento Cruzado Anterior/imunologia , Cartilagem Articular/imunologia , Citocinas/genética , Modelos Animais de Doenças , Humanos , Masculino , Obesidade/metabolismo , Osteoartrite/etiologia , Osteoartrite/genética , Osteoartrite/imunologia , Ratos , Ratos Sprague-Dawley
15.
Mol Cell Biochem ; 461(1-2): 183-193, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31435813

RESUMO

Osteoarthritis (OA) has been identified to be one of the most prevalent forms of joint disorders, marked with inflammatory immune response that may give rise to several complications including disability. Numbers of investigations have proven that microRNA play a key role in chondrogenesis regulation. Accordingly, the current study was intended to explore more about the potential role of miR-940 in the regulation of immune response, pertaining to osteoarthritis. Our findings indicated miR-940 associated down-regulation in both, the tissue as well as at cellular levels, i.e. chondrocytes that are being induced with IL-1ß. However, the expression of MyD88 was found to be opposite. Moreover, our findings indicated that miR-940 targets MyD88 to regulate its expression. The study was based on the proposition that normal human chondrocytes when induced with IL-1ß significantly enhanced the level of inflammation along with simultaneous stimulation of NF-κB signaling mechanism. Alternatively, siRNA against MyD88, miR-940 mimic or the NF-κB inhibitor, reversed the effect of IL-1ß. The chondrocytes that were transfected with miR-940 inhibitor increased the secretion of inflammatory cytokines and activated NF-κB. Furthermore, the expression of miR-490 was reduced in vivo, which was increased through an injection of lentivirus, to foster the production of necessary cytokines and NF-κB and the down-regulation of MyD88. In conclusion, the pathogenesis of OA can be regulated by miR-940/MyD88 axis, which can be achieved through the combined signaling mechanism of MyD88/NF-κB signaling, induced with the help of IL-1ß.


Assuntos
Condrócitos/metabolismo , Condrócitos/patologia , Inflamação/genética , MicroRNAs/metabolismo , Fator 88 de Diferenciação Mieloide/metabolismo , Osteoartrite/genética , Regiões 3' não Traduzidas/genética , Adulto , Idoso , Animais , Sequência de Bases , Proliferação de Células , Citocinas/metabolismo , Feminino , Regulação da Expressão Gênica , Humanos , Inflamação/patologia , Interleucina-1beta/metabolismo , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Fator 88 de Diferenciação Mieloide/genética , NF-kappa B/metabolismo , Osteoartrite/patologia , Ratos , Transdução de Sinais
16.
BMC Complement Altern Med ; 19(1): 191, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-31362725

RESUMO

BACKGROUND: Wnt/ß-catenin signaling pathway is closely related to osteoarthritis. In our preliminary study, ß-catenin conditional activation (cAct) mice that specifically over-express ß-catenin gene in cartilage chondrocyte exhibits osteoarthritis-like phenotype in the lumbar disc and knee joint. Therefore, we used the mice to model FJ-OA and test the potential curative effect of Velvet Antler Polypeptide (VAP) on this mice model. METHODS: We tested the effect of VAP on ß-catenin conditional activation mice, and used Cre negative littermates as controls. Micro-CT, histology and histomorphometry analysis were performed to evaluate the curative effect of VAP on mice facet joint-like phenotype. Expression of ß-catenin and collagen II was detected by immunohistochemistry (IHC) and western-blot., MMP13, ADAMTS4 and ADAMTS5 was detected by immunofluorescence (IF). RT-PCR analysis was preformed to detect mRNA expression of cartilage degrading enzymes, such as MMP13, ADAMTS4 and ADAMTS5. RESULTS: Results of micro-CT (µCT) analysis showed that VAP could partially reverse lumbar disc osteophyte formation observed in ß-catenin(ex3)Col2ER mice. Histology data revealed VAP partially improved facet joint cartilage tissue invades. Histomorphometry analysis showed an increase in total cartilage area after VAP treatment. IHC show that VAP reduced ß-catenin protein levels and moderately up-regulated collagen II protein levels. RT-PCR and IF data showed that VAP down-regulated the expression of extracellular matrix synthesis (ECM) degradation enzymes MMP13, ADAMTS4 and ADAMTS5. CONCLUSION: Taken together, VAP may modulate ECM by inhibits MMP13, ADAMTS4 and ADAMTS5 via Wnt /ß-catenin signaling pathway. Velvet Antler Polypeptide may be a potential medicine for FJ-OA.


Assuntos
Chifres de Veado/química , Osteoartrite/tratamento farmacológico , Peptídeos/administração & dosagem , beta Catenina/metabolismo , Proteína ADAMTS4/genética , Proteína ADAMTS4/metabolismo , Proteína ADAMTS5/genética , Proteína ADAMTS5/metabolismo , Animais , Apoptose/efeitos dos fármacos , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Cervos , Humanos , Articulações/efeitos dos fármacos , Articulação do Joelho/efeitos dos fármacos , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Camundongos , Osteoartrite/genética , Osteoartrite/metabolismo , beta Catenina/genética
17.
Med Arch ; 73(3): 144-148, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31402800

RESUMO

Introduction: Previous research found that diabetes mellitus capable to aggravate osteoarthritis disease. In brief, the hyperglycemia condition in diabetes mellitus has an impact on protein glycation of all joint components, including molecule, such as perlecan. The protein expression of perlecan reflects the presence amount of perlecan in the matrix of articular cartilage. However, the impact of hyperglycemia on articular perlecan has not been explained. Moreover, the role of perlecan as a mechanotransducer for chondrocytes in type 1 Diabetes mellitus remains unclear. Aim: This research aims to analyze the effect of hyperglycemia in type 1 Diabetes mellitus to the mRNA level and protein expression of perlecan. Methods: Thirty-five adult male rats were divided into seven groups, such as three groups of rat model with anterior cruciate ligament transection (ACLT) at right knee (ACLT1, ACLT2, ACLT3); three groups of rats with ACLT at right knee which followed by Streptozotocin injection for diabetic mice model (DM1, DM2, DM3); and control group (N). Rat sacrificed at the third week, fourth week, and sixth week after two months of maintenance. The mRNA level and protein expression were analyzed by using PCR and Western blot. All of data was analyzed by ANOVA. Results: Protein expression of perlecan in ACLT mice with diabetes mellitus (DM1, DM2, DM3 group) was gradually decreased according to the increased hyperglycemia duration. Whilst, protein expression of perlecan within ACLT mice without diabetes mellitus (ACLT1, ACLT2, ACLT3 group) was increased. The similar result also demonstrated by the mRNA level of perlecan. Group of DM1, DM2, DM3 exhibited decreased mRNA level of perlecan over the hyperglycemia duration. While, ACLT1, ACLT2, and ACLT3 group had a gradually increased of perlecan mRNA level. Conclusion: Hyperglycemia on osteoarthritic condition decreased mRNA level and protein expression of perlecan which increased the severity of osteoarthritis disease.


Assuntos
Diabetes Mellitus Experimental/sangue , Diabetes Mellitus Tipo 1/sangue , Proteoglicanas de Heparan Sulfato/genética , Proteoglicanas de Heparan Sulfato/metabolismo , Hiperglicemia/sangue , Osteoartrite/metabolismo , Animais , Glicemia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Tipo 1/complicações , Modelos Animais de Doenças , Hiperglicemia/etiologia , Masculino , Osteoartrite/complicações , Osteoartrite/genética , RNA Mensageiro/metabolismo , Ratos
18.
Food Funct ; 10(9): 5873-5885, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31464310

RESUMO

Osteoarthritis (OA) is the most prevalent form of human arthritis which is characterized by the degradation of cartilage and inflammation. As a rare Sirt6 activator, cyanidin is the major component of anthocyanins commonly found in the Mediterranean diet, and increasing evidence has shown that cyanidin exhibits anti-inflammatory effects in a variety of diseases. However, the anti-inflammatory effects of cyanidin on OA have not been reported. In the present study, we identified that cyanidin treatment could strongly suppress the expression of NO, PGE2, TNF-α, IL-6, iNOs, COX-2, ADAMTS5 and MMP13, and reduce the degradation of aggrecan and collagen II in IL-1ß-induced human OA chondrocytes, indicating the anti-inflammatory effect of cyanidin. Further investigation of the mechanism involved revealed that cyanidin could upregulate the Sirt6 level in a dose-dependent manner and Sirt6 silencing abolished the effect of cyanidin in IL-1ß-stimulated human OA chondrocytes, indicating a stimulatory effect of cyanidin on Sirt6 activation. Meanwhile, we found that cyanidin could inhibit the NF-κB pathway in IL-1ß-stimulated human OA chondrocytes and its effect may to some extent depend on Sirt6 activation, suggesting that cyanidin may exert a protective effect through regulating the Sirt6/NF-κB signaling axis. Moreover, the in vivo study also proved that cyanidin ameliorated the development of OA in surgical destabilization of the medial meniscus (DMM) mouse OA models. In conclusion, these results demonstrate that cyanidin may have therapeutic potential for the treatment of OA.


Assuntos
Antocianinas/administração & dosagem , Anti-Inflamatórios/administração & dosagem , NF-kappa B/imunologia , Osteoartrite/tratamento farmacológico , Sirtuínas/imunologia , Animais , Condrócitos/efeitos dos fármacos , Condrócitos/imunologia , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/imunologia , Humanos , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Masculino , Metaloproteinase 13 da Matriz , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Osteoartrite/genética , Osteoartrite/imunologia , Sirtuínas/genética
19.
EBioMedicine ; 47: 402-413, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31466915

RESUMO

BACKGROUND: Although neoangiogenesis is a hallmark of chronic inflammatory diseases such as inflammatory arthritis and many cancers, therapeutic agents targeting the vasculature remain elusive. Here we identified miR-125a as an important regulator of angiogenesis. METHODS: MiRNA levels were quantified in Psoriatic Arthritis (PsA) synovial-tissue by RT-PCR and compared to macroscopic synovial vascularity. HMVEC were transfected with anti-miR-125a and angiogenic mechanisms quantified using tube formation assays, transwell invasion chambers, wound repair, RT-PCR and western blot. Real-time analysis of EC metabolism was assessed using the XF-24 Extracellular-Flux Analyzer. Synovial expression of metabolic markers was assessed by immunohistochemistry and immunofluorescent staining. MiR-125a CRISPR/Cas9-based knock-out zebrafish were generated and vascular development assessed. Finally, glycolytic blockade using 3PO, which inhibits Phosphofructokinase-fructose-2,6-bisphophatase 3 (PFKFB3), was assessed in miR-125a-/- ECs and zebrafish embryos. FINDINGS: MiR-125a is significantly decreased in PsA synovium and inversely associated with macroscopic vascularity. In-vivo, CRISPR/cas9 miR-125a-/- zebrafish displayed a hyper-branching phenotype. In-vitro, miR-125a inhibition promoted EC tube formation, branching, migration and invasion, effects paralleled by a shift in their metabolic profile towards glycolysis. This metabolic shift was also observed in the PsA synovial vasculature where increased expression of glucose transporter 1 (GLUT1), PFKFB3 and Pyruvate kinase muscle isozyme M2 (PKM2) were demonstrated. Finally, blockade of PFKFB3 significantly inhibited anti-miR-125a-induced angiogenic mechanisms in-vitro, paralleled by normalisation of vascular development of CRISPR/cas9 miR-125a-/- zebrafish embryos. INTEPRETATION: Our results provide evidence that miR-125a deficiency enhances angiogenic processes through metabolic reprogramming of endothelial cells. FUND: Irish Research Council, Arthritis Ireland, EU Seventh Framework Programme (612218/3D-NET).


Assuntos
Regulação da Expressão Gênica , MicroRNAs/genética , Neovascularização Patológica/genética , Animais , Biópsia , Movimento Celular , Proliferação de Células , Modelos Animais de Doenças , Células Endoteliais , Inativação Gênica , Glicólise , Humanos , Osteoartrite/genética , Osteoartrite/patologia , Interferência de RNA , Peixe-Zebra
20.
Biomed Res Int ; 2019: 7165406, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31467907

RESUMO

Early detection and treatment are critical in the management of osteoarthritis (OA). OA is closely associated with angiogenesis and the inhibition of angiogenesis presents a novel therapeutic approach to reduce inflammation and pain in OA. Recent reports suggest that circulating microRNAs (miRNAs) have great potential as biomarkers for the diagnosis and prognosis in OA. In this study, we aimed to explore the clinical significance of miR-210 in synovial fluid samples from 10 healthy volunteers and 20 early-stage OA and 20 late-stage OA patients. miR-210 expression was assessed by real-time RT-PCR. VEGF protein levels were examined by ELISA. The results show that miR-210 is significantly upregulated in early-stage OA and late-stage OA patients compared with healthy individuals. Higher levels of VEGF are also found in OA compared with the control. Moreover, miR-210 levels are positively correlated with VEGF levels, suggesting that miR-210 might contribute to OA development through promoting VEGF expression and angiogenesis. In conclusion, upregulation of miR-210 in synovial fluid may occur in the early stage of OA and can be a useful biomarker for early diagnosis of OA.


Assuntos
Biomarcadores/metabolismo , Diagnóstico Precoce , MicroRNAs/genética , Osteoartrite/diagnóstico , Idoso , MicroRNA Circulante , Feminino , Regulação da Expressão Gênica/genética , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite/genética , Osteoartrite/patologia , Líquido Sinovial/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA