Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 2.872
Filtrar
1.
Int J Mol Sci ; 22(11)2021 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-34073090

RESUMO

Nuclear magnetic resonance therapy (NMRT) is discussed as a participant in repair processes regarding cartilage and as an influence in pain signaling. To substantiate the application of NMRT, the underlying mechanisms at the cellular level were studied. In this study microRNA (miR) was extracted from human primary healthy and osteoarthritis (OA) chondrocytes after NMR treatment and was sequenced by the Ion PI Hi-Q™ Sequencing 200 system. In addition, T/C-28a2 chondrocytes grown under hypoxic conditions were studied for IL-1ß induced changes in expression on RNA and protein level. HDAC activity an NAD(+)/NADH was measured by luminescence detection. In OA chondrocytes miR-106a, miR-27a, miR-34b, miR-365a and miR-424 were downregulated. This downregulation was reversed by NMRT. miR-365a-5p is known to directly target HDAC and NF-ĸB, and a decrease in HDAC activity by NMRT was detected. NAD+/NADH was reduced by NMR treatment in OA chondrocytes. Under hypoxic conditions NMRT changed the expression profile of HIF1, HIF2, IGF2, MMP3, MMP13, and RUNX1. We conclude that NMRT changes the miR profile and modulates the HDAC and the NAD(+)/NADH signaling in human chondrocytes. These findings underline once more that NMRT counteracts IL-1ß induced changes by reducing catabolic effects, thereby decreasing inflammatory mechanisms under OA by changing NF-ĸB signaling.


Assuntos
Condrócitos , Espectroscopia de Ressonância Magnética/métodos , MicroRNAs/metabolismo , Osteoartrite , Linhagem Celular , Condrócitos/citologia , Condrócitos/metabolismo , Condrócitos/patologia , Humanos , Inflamação/metabolismo , Inflamação/patologia , Osteoartrite/metabolismo , Osteoartrite/terapia , Cultura Primária de Células
2.
Life Sci ; 278: 119553, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33932445

RESUMO

AIMS: We have evaluated the potential of a three-dimensional (3D) thermoreversible gelation polymer (TGP) matrix in enhancing miRNA 140 expression (a biomarker correlating with homeostasis and cartilage regeneration) during the in vitro expansion of osteoarthritis (OA)-affected human chondrocytes. MATERIALS AND METHODS: OA-chondrocytes were cultured in two-dimensional (2D) monolayer followed by culture in 3D-TGP. miRNA 140 expression levels in cell culture supernatant followed by expression in the cell lysate of both 2D and 3D-TGP cultures were analyzed. KEY FINDINGS: The expression of miRNA 140 in cell culture supernatant from the 3D-TGP group was 0.001 to 0.002% that in 2D culture supernatant while in the cell lysate, miRNA 140 expression in the 3D-TGP was nearly 30-fold higher than that of 2D group. SIGNIFICANCE: The 3D-TGP matrix allows enhanced expression of miRNA 140 in OA-affected human chondrocytes in vitro which after necessary validations can be applied in clinical transplantation to significantly improve the outcome.


Assuntos
Condrócitos/patologia , MicroRNAs/genética , Osteoartrite/genética , Regulação para Cima , Técnicas de Cultura de Células , Células Cultivadas , Condrócitos/metabolismo , Humanos , Osteoartrite/patologia , Polímeros/química , Tecidos Suporte/química
3.
Int J Mol Sci ; 22(5)2021 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-33803113

RESUMO

Obesity is a risk factor for osteoarthritis (OA) development and progression due to an altered biomechanical stress on cartilage and an increased release of inflammatory adipokines from adipose tissue. Evidence suggests an interplay between loading and adipokines in chondrocytes metabolism modulation. We investigated the role of loading, as hydrostatic pressure (HP), in regulating visfatin-induced effects in human OA chondrocytes. Chondrocytes were stimulated with visfatin (24 h) and exposed to high continuous HP (24 MPa, 3 h) in the presence of visfatin inhibitor (FK866, 4 h pre-incubation). Apoptosis and oxidative stress were detected by cytometry, B-cell lymphoma (BCL)2, metalloproteinases (MMPs), type II collagen (Col2a1), antioxidant enzymes, miRNA, cyclin D1 expressions by real-time PCR, and ß-catenin protein by western blot. HP exposure or visfatin stimulus significantly induced apoptosis, superoxide anion production, and MMP-3, -13, antioxidant enzymes, and miRNA gene expression, while reducing Col2a1 and BCL2 mRNA. Both stimuli significantly reduced ß-catenin protein and increased cyclin D1 gene expression. HP exposure exacerbated visfatin-induced effects, which were counteracted by FK866 pre-treatment. Our data underline the complex interplay between loading and visfatin in controlling chondrocytes' metabolism, contributing to explaining the role of obesity in OA etiopathogenesis, and confirming the importance of controlling body weight for disease treatment.


Assuntos
Adipocinas/biossíntese , Apoptose , Condrócitos/metabolismo , Regulação da Expressão Gênica , Osteoartrite/metabolismo , Idoso , Células Cultivadas , Condrócitos/patologia , Feminino , Humanos , Pressão Hidrostática , Masculino , Pessoa de Meia-Idade , Nicotinamida Fosforribosiltransferase/farmacologia , Osteoartrite/patologia
4.
Int J Mol Sci ; 22(5)2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33804447

RESUMO

Interleukin (IL)-1ß is an important pro-inflammatory cytokine in the progression of osteoarthritis (OA), which impairs mitochondrial function and induces the production of nitric oxide (NO) in chondrocytes. The aim was to investigate if blockade of NO production prevents IL-1ß-induced mitochondrial dysfunction in chondrocytes and whether cAMP and AMP-activated protein kinase (AMPK) affects NO production and mitochondrial function. Isolated human OA chondrocytes were stimulated with IL-1ß in combination with/without forskolin, L-NIL, AMPK activator or inhibitor. The release of NO, IL-6, PGE2, MMP3, and the expression of iNOS were measured by ELISA or Western blot. Parameters of mitochondrial respiration were measured using a seahorse analyzer. IL-1ß significantly induced NO release and mitochondrial dysfunction. Inhibition of iNOS by L-NIL prevented IL-1ß-induced NO release and mitochondrial dysfunction but not IL-1ß-induced release of IL-6, PGE2, and MMP3. Enhancement of cAMP by forskolin reduced IL-1ß-induced NO release and prevented IL-1ß-induced mitochondrial impairment. Activation of AMPK increased IL-1ß-induced NO production and the negative impact of IL-1ß on mitochondrial respiration, whereas inhibition of AMPK had the opposite effects. NO is critically involved in the IL-1ß-induced impairment of mitochondrial respiration in human OA chondrocytes. Increased intracellular cAMP or inhibition of AMPK prevented both IL-1ß-induced NO release and mitochondrial dysfunction.


Assuntos
Condrócitos/efeitos dos fármacos , Inflamação/prevenção & controle , Interleucina-1beta/farmacologia , Mitocôndrias/efeitos dos fármacos , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico/metabolismo , Osteoartrite do Joelho/prevenção & controle , Células Cultivadas , Condrócitos/metabolismo , Condrócitos/patologia , Feminino , Humanos , Inflamação/etiologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Pessoa de Meia-Idade , Mitocôndrias/metabolismo , Mitocôndrias/patologia , NF-kappa B/genética , NF-kappa B/metabolismo , Osteoartrite do Joelho/induzido quimicamente , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia
5.
Int J Mol Sci ; 22(6)2021 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-33799449

RESUMO

(1) Background: Tissue non-specific alkaline phosphatase (TNAP) is suspected to induce atherosclerosis plaque calcification. TNAP, during physiological mineralization, hydrolyzes the mineralization inhibitor inorganic pyrophosphate (PPi). Since atherosclerosis plaques are characterized by the presence of necrotic cells that probably release supraphysiological concentrations of ATP, we explored whether this extracellular adenosine triphosphate (ATP) is hydrolyzed into the mineralization inhibitor PPi or the mineralization stimulator inorganic phosphate (Pi), and whether TNAP is involved. (2) Methods: Murine aortic smooth muscle cell line (MOVAS cells) were transdifferentiated into chondrocyte-like cells in calcifying medium, containing ascorbic acid and ß-glycerophosphate. ATP hydrolysis rates were determined in extracellular medium extracted from MOVAS cultures during their transdifferentiation, using 31P-NMR and IR spectroscopy. (3) Results: ATP and PPi hydrolysis by MOVAS cells increased during transdifferentiation. ATP hydrolysis was sequential, yielding adenosine diphosphate (ADP), adenosine monophosphate (AMP), and adenosine without any detectable PPi. The addition of levamisole partially inhibited ATP hydrolysis, indicating that TNAP and other types of ectonucleoside triphoshatediphosphohydrolases contributed to ATP hydrolysis. (4) Conclusions: Our findings suggest that high ATP levels released by cells in proximity to vascular smooth muscle cells (VSMCs) in atherosclerosis plaques generate Pi and not PPi, which may exacerbate plaque calcification.


Assuntos
Aterosclerose/genética , Transdiferenciação Celular/genética , Difosfatos/metabolismo , Calcificação Vascular/genética , Trifosfato de Adenosina , Fosfatase Alcalina/genética , Animais , Aorta/citologia , Aorta/metabolismo , Ácido Ascórbico/farmacologia , Aterosclerose/metabolismo , Aterosclerose/patologia , Condrócitos/metabolismo , Condrócitos/patologia , Glicerofosfatos/genética , Glicerofosfatos/metabolismo , Humanos , Espectroscopia de Ressonância Magnética , Camundongos , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/patologia , Fosfatos/metabolismo , Calcificação Vascular/metabolismo , Calcificação Vascular/patologia
6.
Aging (Albany NY) ; 13(8): 11646-11664, 2021 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-33879632

RESUMO

OBJECTIVE: To investigate the heterogeneous responses of in vitro expanded chondrocytes, which were cultured in an interleukin (IL)-1ß -induced inflammatory environment. METHOD: Human articular chondrocytes were expanded, in vitro, for 13 days and treated with IL-1ß for 0, 24, and 48 h. Cells were collected and subjected to single-cell RNA sequencing. Multiple bioinformatics tools were used to determine the signatures that define chondrocyte physiology. RESULTS: Two major cell clusters with distinct expression patterns were identified at the initial phase and were with heterogeneous variation that coincides with inflammation progress. They transformed into two terminal cell clusters one of which exhibited OA-phenotype and proinflammatory characteristics through two paths, "response-to-inflammation" and "atypical response-to-inflammation", respectively. The involved cell clusters exhibited intrinsic relationship with cell types within native cartilage from OA patients. Genes controlling cell transformation to OA-phenotype were relating to the tumor necrosis factor (TNF) signaling pathway via NFKB, up-regulated KRAS signaling and the IL2/STAT5 signaling pathway and pathways relating to apoptosis and reactive oxygen species. CONCLUSION: The in vitro expanded chondrocytes under IL-1ß-induced inflammatory progression behave heterogeneously. One of the initial cell clusters could transform into a proinflammatory subpopulation through a termed response-to-inflammation path, which may serve as the core target to alleviate OA progression.


Assuntos
Condrócitos/patologia , Regulação da Expressão Gênica/imunologia , Osteoartrite/imunologia , Transdução de Sinais/genética , Cartilagem Articular/citologia , Células Cultivadas , Criança , Condrócitos/imunologia , Biologia Computacional , Meios de Cultura/metabolismo , Humanos , Interleucina-1beta/metabolismo , Osteoartrite/genética , Osteoartrite/patologia , Cultura Primária de Células , RNA-Seq , Transdução de Sinais/imunologia , Análise de Célula Única
7.
Methods Mol Biol ; 2269: 221-231, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33687682

RESUMO

Adipose tissue-derived mesenchymal stem cells (AD-MSCs) offer great therapeutic potential for osteoarthritis (OA) treatment. Recent investigations have revealed the contribution of extracellular vesicles (EVs) to AD-MSC actions. Here, we describe a method to study the in vitro effects of EVs from AD-MSCs in OA chondrocytes. This chapter includes the isolation and analysis of human AD-MSCs and their EVs as well as the isolation and treatment of OA chondrocytes.


Assuntos
Tecido Adiposo/metabolismo , Condrócitos/metabolismo , Vesículas Extracelulares/metabolismo , Células-Tronco Mesenquimais/metabolismo , Osteoartrite/metabolismo , Tecido Adiposo/patologia , Separação Celular , Condrócitos/patologia , Técnicas de Cocultura , Humanos , Células-Tronco Mesenquimais/patologia , Osteoartrite/patologia
8.
Life Sci ; 274: 119324, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33711382

RESUMO

AIMS: Intervertebral Disc Degeneration (IDD) is a key factor involved in low back pain (LBP) which affects approximately 540 million individuals worldwide. Chlorogenic Acid (CGA), a natural compound, exerts anti-inflammatory property in several diseases. Here, we aim to investigate the biological effect of CGA on IDD and explore the underlying mechanism. MATERIALS AND METHODS: Lumbar spine instability (LSI) model in mice was utilized to mimic process of IDD. The effects of CGA in response to LSI were evaluated by luminescent imaging, micro-CT, histomorphology, and immunohistochemistry in vivo. Besides, the cytotoxicity of CGA on chondrocytes was detected by cell counting kit-8 (CCK-8) and the biological effects were assessed by polymerase chain reaction (PCR) in vitro. KEY FINDINGS: We found that CGA treatment dramatically suppressed the NF-κB activity in LSI mice. Moreover, administration of CGA mitigated cartilaginous endplate degeneration and postponed IDD development accompanying a decrease of inflammatory and catabolic mediators. Specifically, CGA ameliorated endplate degeneration might be related to its protective effects against endplate chondrocytes apoptosis and trans-differentiation. We further elucidated that CGA exerted these biological effects mainly by repressing NF-κB signaling in cartilage endplate. SIGNIFICANCE: Our study has illustrated, for the first time, the curative effects as well as the latent mechanism of CGA in IDD and our results suggested that CGA administration might be used as an alternative therapy for IDD.


Assuntos
Apoptose , Cartilagem/efeitos dos fármacos , Ácido Clorogênico/farmacologia , Condrócitos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Degeneração do Disco Intervertebral/tratamento farmacológico , NF-kappa B/antagonistas & inibidores , Animais , Cartilagem/patologia , Células Cultivadas , Condrócitos/patologia , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL
9.
Cell Mol Life Sci ; 78(7): 3743-3762, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33683377

RESUMO

Mutations in the intraflagellar transport-A (IFT-A) gene, THM1, have been identified in skeletal ciliopathies. Here, we report a genetic interaction between Thm1, and its paralog, Thm2, in postnatal skeletogenesis. THM2 localizes to primary cilia, but Thm2 deficiency does not affect ciliogenesis and Thm2-null mice survive into adulthood. However, by postnatal day 14, Thm2-/-; Thm1aln/+ mice exhibit small stature and small mandible. Radiography and microcomputed tomography reveal Thm2-/-; Thm1aln/+ tibia are less opaque and have reduced cortical and trabecular bone mineral density. In the mutant tibial growth plate, the proliferation zone is expanded and the hypertrophic zone is diminished, indicating impaired chondrocyte differentiation. Additionally, mutant growth plate chondrocytes show increased Hedgehog signaling. Yet deletion of one allele of Gli2, a major transcriptional activator of the Hedgehog pathway, exacerbated the Thm2-/-; Thm1aln/+ small phenotype, and further revealed that Thm2-/-; Gli2+/- mice have small stature. In Thm2-/-; Thm1aln/+ primary osteoblasts, a Hedgehog signaling defect was not detected, but bone nodule formation was markedly impaired. This indicates a signaling pathway is altered, and we propose that this pathway may potentially interact with Gli2. Together, our data reveal that loss of Thm2 with one allele of Thm1, Gli2, or both, present new IFT mouse models of osteochondrodysplasia. Our data also suggest Thm2 as a modifier of Hedgehog signaling in postnatal skeletal development.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/fisiologia , Condrócitos/patologia , Condrogênese , Proteínas Hedgehog/metabolismo , Osteoblastos/patologia , Osteogênese , Animais , Animais Recém-Nascidos , Diferenciação Celular , Condrócitos/metabolismo , Cílios , Feminino , Proteínas Hedgehog/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Osteoblastos/metabolismo , Transdução de Sinais
10.
Int J Mol Sci ; 22(5)2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33668140

RESUMO

Osteoarthritis (OA) is a slow-progressing joint disease, leading to the degradation and remodeling of the cartilage extracellular matrix (ECM). The usually quiescent chondrocytes become reactivated and accumulate in cell clusters, become hypertrophic, and intensively produce not only degrading enzymes, but also ECM proteins, like the cartilage oligomeric matrix protein (COMP) and thrombospondin-4 (TSP-4). To date, the functional roles of these newly synthesized proteins in articular cartilage are still elusive. Therefore, we analyzed the involvement of both proteins in OA specific processes in in vitro studies, using porcine chondrocytes, isolated from femoral condyles. The effect of COMP and TSP-4 on chondrocyte migration was investigated in transwell assays and their potential to modulate the chondrocyte phenotype, protein synthesis and matrix formation by immunofluorescence staining and immunoblot. Our results demonstrate that COMP could attract chondrocytes and may contribute to a repopulation of damaged cartilage areas, while TSP-4 did not affect this process. In contrast, both proteins similarly promoted the synthesis and matrix formation of collagen II, IX, XII and proteoglycans, but inhibited that of collagen I and X, resulting in a stabilized chondrocyte phenotype. These data suggest that COMP and TSP-4 activate mechanisms to protect and repair the ECM in articular cartilage.


Assuntos
Artrite Experimental/metabolismo , Proteína de Matriz Oligomérica de Cartilagem/metabolismo , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Matriz Extracelular/metabolismo , Osteoartrite/metabolismo , Trombospondinas/metabolismo , Animais , Artrite Experimental/patologia , Cartilagem Articular/patologia , Condrócitos/patologia , Feminino , Osteoartrite/patologia , Suínos
11.
Carbohydr Polym ; 261: 117869, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-33766356

RESUMO

Osteoarthritis (OA) is an age-related joint disorder and one of the leading causes of physical disability. In this study, we designed and synthesized a new polysaccharide complex, carboxymethyl chitosan strontium (CMCS-Sr), which is believed to have positive effects on relieving OA. The synthesized CMCS-Sr was structurally verified by SEM, EDS, FTIR, etc. The therapeutic effects of CMCS-Sr were evaluated using various biological experiments. The cell viability and apoptosis results reveal that CMCS-Sr can significantly promote the proliferation and suppress OA chondrocytes apoptosis in vitro. The immunofluorescence staining results suggest that CMCS-Sr facilitates the promotion of the secretion of Type II collagen (Col-II). The transcriptomic results support the observed positive effects of CMCS-Sr on inhibiting chondrocytes apoptosis and alleviating inflammatory reactions. Moreover, animal study demonstrates that CMCS-Sr effectively reduced articular cartilage damage and subchondral bone degradation. Therefore, we propose the use of CMCS-Sr as a promising candidate for relieving OA.


Assuntos
Quitosana/análogos & derivados , Osteoartrite/tratamento farmacológico , Polímeros/síntese química , Polímeros/uso terapêutico , Estrôncio/química , Animais , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/patologia , Células Cultivadas , Quitosana/síntese química , Quitosana/química , Quitosana/farmacologia , Quitosana/uso terapêutico , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Condrócitos/fisiologia , Humanos , Masculino , Osteoartrite/patologia , Polímeros/química , Polímeros/farmacologia , Cultura Primária de Células , Ratos , Estrôncio/farmacologia , Estrôncio/uso terapêutico
12.
Life Sci ; 275: 119375, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33737085

RESUMO

AIMS: Both aerobic exercise and glucosamine hydrochloride capsules (OTL) have a therapeutic effect on knee osteoarthritis, but their joint application has not been investigated. This study clarified the mechanism of the combined treatment in knee osteoarthritis. MAIN METHODS: Aerobic exercise and OTL were used alone or in combination to treat papain-induced knee osteoarthritis model rabbits. Pathological changes of cartilage tissues, inflammatory cytokine content, glycosaminoglycan, and expressions of collagen II, cartilage differentiation-related genes and circUNK were analyzed by hematoxylin-eosin staining, Mankin score, Enzyme-linked immunosorbent assay, toluidine blue staining, Immunohistochemistry and qRT-PCR. The extracted chondrocytes were identified by Alcian Blue staining and immunohistochemistry and induced by iodoacetic acid (MIA) to establish osteoarthritis model. Effects of overexpressing or silencing circUNK on cell function and molecular changes in chondrocytes were analyzed by cell function experiments, qRT-PCR and Western blot. Rabbit modeling and intervention treatment were marked. KEY FINDINGS: Aerobic exercise or OTL treatment alone relieved the damage caused by knee osteoarthritis in terms of cartilage tissue lesions, Mankin score, inflammatory cytokine content, glycosaminoglycan, and expressions of collagen II, cartilage differentiation-related genes and circUNK. Combined application of aerobic exercise and OTL showed better synergistic treatment effects. Transfection of overexpressed circUNK could attenuate the MIA-induced effect on cell viability and apoptosis in chondrocytes by regulating genes related to differentiation and apoptosis. Aerobic exercise combined with glucosamine had a synergistic therapeutic effect on knee osteoarthritis. SIGNIFICANCE: Overexpressing circUNK protected osteoarthritis model cells by regulating cartilage differentiation- and apoptosis-related genes.


Assuntos
Glucosamina/uso terapêutico , Osteoartrite do Joelho/terapia , Condicionamento Físico Animal , RNA Circular/metabolismo , Animais , Western Blotting , Cartilagem Articular/patologia , Condrócitos/patologia , Terapia Combinada , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Osteoartrite do Joelho/patologia , Condicionamento Físico Animal/métodos , Coelhos , Reação em Cadeia da Polimerase em Tempo Real
13.
Biochem Biophys Res Commun ; 553: 119-125, 2021 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-33765556

RESUMO

Excessive activation of inflammation in chondrocyte has been considered to be a major reason cause of cellular death and degeneration in osteoarthritis (OA) development. The NLRP3 inflammasome-mediated pyroptosis pathway is closely related to inflammation regulation. This research was conducted to confirm whether NLRP3 expression and activity are impacted in the development of OA and to detect the role of CY-09, a selective and direct inhibitor of NLRP3 in the in vitro and in vivo models of OA. Our findings corroborated that the expression of NLRP3 is stimulated in OA cartilage. CY-09 can maintain extracellular matrix (ECM) homeostasis and regulate inflammation in TNF-α treated chondrocytes via inhibition of NLRP3 inflammasome-mediated pyroptosis. Moreover, the chondrocyte protective effects of CY-09 were further confirmed in vivo in a DMM-induced OA model. In conclusion, our research indicates that experimental OA activated the NLRP3 activity, and pharmacological inhibition of NLRP3 inflammasome activation by CY-09 protects chondrocytes against inflammation and attenuates OA development.


Assuntos
Inflamassomos/antagonistas & inibidores , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Osteoartrite/tratamento farmacológico , Substâncias Protetoras/farmacologia , Piroptose/efeitos dos fármacos , Tiazolidinas/farmacologia , Tionas/farmacologia , Animais , Cartilagem/efeitos dos fármacos , Cartilagem/metabolismo , Cartilagem/patologia , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Modelos Animais de Doenças , Progressão da Doença , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/patologia , Feminino , Homeostase/efeitos dos fármacos , Inflamassomos/metabolismo , Inflamação/induzido quimicamente , Inflamação/patologia , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Osteoartrite/patologia , Osteoartrite/prevenção & controle , Fator de Necrose Tumoral alfa/farmacologia
14.
Acta Biochim Biophys Sin (Shanghai) ; 53(4): 400-409, 2021 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-33677475

RESUMO

Persistent hypotonic and inflammatory conditions in the joint cavity can lead to the loss of cartilage matrix and cell death, which are the important mechanisms of osteoarthritis (OA) onset. Previous studies have confirmed that the existence of a hypotonic environment is a red flag for inflammation, as hypotonic environment induces the opening of the chloride channel of the cell and promotes chloride ion efflux, which prompts the cell volume to increase. Chloride channels play an important role in the regulation of mineralization and chondrocyte death. Here, we reported that OA chondrocytes showed a significant increase of cell death rate and the imbalance of cartilage matrix catabolism. We found that the distribution of skeleton protein F-actin was disordered. In addition, the volume-sensitive chloride current of OA chondrocytes decreased significantly with the increase of the expression levels of inflammation-related proteins caspase-1, caspase-3, and NLRP3. Moreover, interleukin-1ß (IL-1ß) showed a potential to activate the chloride current of normal chondrocytes. These results indicate that IL-1ß-induced chloride channel opening in chondrocytes may be closely related to the occurrence of OA. This chloride channel opening process may therefore be a potential target for the treatment of OA.


Assuntos
Cloretos/metabolismo , Condrócitos/metabolismo , Interleucina-1beta/metabolismo , Osteoartrite/metabolismo , Idoso , Idoso de 80 Anos ou mais , Condrócitos/patologia , Feminino , Humanos , Inflamação/metabolismo , Inflamação/patologia , Transporte de Íons , Masculino , Osteoartrite/patologia
15.
Biosci Biotechnol Biochem ; 85(4): 842-850, 2021 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-33686420

RESUMO

Osteoarthritis (OA) seriously affects people's quality of life due to joint pain, stiffness, disability, and dyskinesia worldwide. Long noncoding RNA zinc finger antisense 1 (ZFAS1) is downregulated and tightly associated with proliferation, migration, apoptosis, and matrix synthesis of chondrocyte in OA. However, the molecular mechanisms of ZFAS1 in OA remain unknown. The expression correlation between ZFAS1, miR-302d-3p, and SMAD2 in OA tissues was analyzed by Pearson correlation analysis. ZFAS1 was a lower expression, and expedited proliferation and repressed apoptosis of chondrocytes. MiR-302d-3p was a direct target of ZFAS1. MiR-302d-3p hindered proliferation and facilitated apoptosis of chondrocytes. MiR-302d-3p partially reversed the effect of ZFAS1 on proliferation and apoptosis of chondrocytes. SMAD2 was positively regulated by the ZFAS1/miR-302d-3p. MiR-302d-3p-mediated proliferation and apoptosis were partly abrogated by targeting SMAD2. ZFAS1 promoted chondrocytes proliferation and repressed apoptosis possibly by regulating miR-302d-3p/SMAD2 axis, providing a potential target for OA treatment.


Assuntos
Apoptose/fisiologia , Condrócitos/patologia , MicroRNAs/fisiologia , Osteoartrite/patologia , RNA Longo não Codificante/fisiologia , Proteína Smad2/fisiologia , Proliferação de Células/fisiologia , Humanos
16.
J Orthop Surg (Hong Kong) ; 29(1): 23094990211000168, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33729061

RESUMO

PURPOSE: This study aimed to investigate how fibroblastic and chondrocytic properties of human meniscal fibrochondrocytes are affected in culture conditions according to the type of meniscal pathology and localization, and to provide basic information for tissue-engineering studies. METHODS: Primary fibrochondrocyte cultures were prepared from meniscus samples of patients who had either traumatic tear or degeneration due to osteoarthritis. Cultures were compared in terms of mRNA expression levels of COL1A1, COL2A1, COMP1, HIF1A, HIF2A, and SOX9 and secreted total collagen and sulfated sGAG levels according to the type of meniscal pathology, anatomical localization, and the number of subcultures. RESULTS: mRNA expression levels of COL1A1, COMP1, HIF1A, HIF2A, and SOX9 were found to be increased in subsequent subcultures in all specimens. COL1A1 mRNA expression levels of both lateral and medial menisci of patients with traumatic tear were significantly higher than in patients with degenerative pathology, indicating a more fibroblastic character. P1 subculture of lateral and P3 or further subculture of medial meniscus showed more fibroblastic characteristics in patients with degenerative pathology. Furthermore, in patients with degenerative pathology, the subcultures of the lateral meniscus (especially on the inner part) presented more chondrocytic characteristics than did those of medial meniscus. CONCLUSIONS: The mRNA expression levels of the cultures showed significant differences according to the anatomical localization and pathology of the meniscus, indicating distinct chondrocytic and fibroblastic features. This fundamental knowledge would help researchers to choose more efficient cell sources for cell-seeding of a meniscus scaffold, and to generate a construct resembling the original meniscus tissue.


Assuntos
Fibrocartilagem , Articulações/lesões , Menisco , Osteoartrite/patologia , Transcriptoma , Adolescente , Adulto , Idoso , Células Cultivadas , Condrócitos/citologia , Condrócitos/metabolismo , Condrócitos/patologia , Feminino , Fibrocartilagem/citologia , Fibrocartilagem/metabolismo , Fibrocartilagem/patologia , Perfilação da Expressão Gênica , Humanos , Articulações/metabolismo , Articulações/patologia , Masculino , Menisco/citologia , Menisco/lesões , Menisco/metabolismo , Menisco/patologia , Pessoa de Meia-Idade , Osteoartrite/genética , Osteoartrite/metabolismo , Cultura Primária de Células/métodos , Ruptura/genética , Ruptura/metabolismo , Ruptura/patologia , Adulto Jovem
17.
Biosci Biotechnol Biochem ; 85(3): 545-552, 2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33590831

RESUMO

Interleukin-1 receptor-associated kinase-3 (IRAK3) has a distinctive role in regulating inflammation. However, the functional role of IRAK3 and regulatory mechanism underlying the pathogenesis of osteoarthritis (OA) remain unclear. Here, we first found that IRAK3 was upregulated, while miR-33b-3p was downregulated in the cartilage of OA patients and IL-1ß-induced CHON-001 cells. IRAK3 was confirmed as the direct target of miR-33b-3p and negatively regulated by miR-33b-3p. There was an inverse correlation between IRAK3 mRNA expression and miR-33b-3p expression in OA cartilage tissues. The in vitro functional experiments showed that miR-33b-3p overexpression caused a remarkable increase in viability, a significant decrease in inflammatory mediators (IL-1ß and TNF-α), and apoptosis in IL-1ß-induced CHON-001 cells. Importantly, IRAK3 knockdown imitated, while overexpression reversed the effects of miR-33b-3p on IL-1ß-induced inflammation and apoptosis in CHON-001 cells. Collectively, miR-33b-3p significantly alleviated IL-1ß-induced inflammation and apoptosis by downregulating IRAK3, which may serve as a promising target for OA.


Assuntos
Apoptose/fisiologia , Condrócitos/patologia , Regulação para Baixo , Quinases Associadas a Receptores de Interleucina-1/fisiologia , Modelos Biológicos , Osteoartrite/patologia , Linhagem Celular , Condrócitos/metabolismo , Humanos , Mediadores da Inflamação/metabolismo , Quinases Associadas a Receptores de Interleucina-1/genética , Interleucina-1beta/metabolismo , MicroRNAs/fisiologia , Osteoartrite/metabolismo , RNA Mensageiro/genética , Fator de Necrose Tumoral alfa/metabolismo
18.
Mol Med Rep ; 23(4)2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33537828

RESUMO

As a chronic degenerative joint disease, the characteristics of osteoarthritis (OA) are degeneration of articular cartilage, subchondral bone sclerosis and bone hyperplasia. It has been reported that microRNA (miR)­186­5p serves a key role in the development of various tumors, such as osteosarcoma, non­small­cell lung cancer cells, glioma and colorectal cancer. The present study aimed to investigate the effect of miR­186­5p in OA. Different concentrations of IL­1ß were used to treat the human chondrocyte cell line CHON­001 to simulate inflammation, and CHON­001 cell injury was assessed by detecting cell viability, apoptosis, caspase-3 activity and the levels of TNF­α, IL­8 and IL­6. Subsequently, reverse transcription­quantitative PCR was performed to measure miR­186­5p expression. The results demonstrated that following IL­1ß treatment, CHON­001 cell viability was suppressed, apoptosis was promoted, the caspase-3 activity was significantly enhanced and the release of TNF­α, IL­8 and IL­6 was increased. In addition, IL­1ß treatment significantly upregulated miR­186­5p expression in CHON­001 cells. It was also identified that MAPK1 was a target gene of miR­186­5p, and was negatively regulated by miR­186­5p. miR­186 inhibitor and MAPK1­small interfering RNA (siRNA) were transfected into CHON­001 cells to investigate the effect of miR­186­5p on CHON­001 cell injury induced by IL­1ß. The results demonstrated that miR­186 inhibitor suppressed the effects of IL­1ß on CHON­001 cells, and these effects were reversed by MAPK1­siRNA. In conclusion, the present results indicated that miR­186­5p could attenuate IL­1ß­induced chondrocyte inflammation damage by increasing MAPK1 expression, suggesting that miR­186­5p may be used as a potential therapeutic target for OA.


Assuntos
Condrócitos/metabolismo , Regulação para Baixo , MicroRNAs/biossíntese , Osteoartrite/metabolismo , Linhagem Celular , Condrócitos/patologia , Citocinas/metabolismo , Humanos , Proteína Quinase 1 Ativada por Mitógeno , Osteoartrite/patologia
19.
Acta Biochim Biophys Sin (Shanghai) ; 53(5): 517-527, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33638344

RESUMO

The homeostasis of the vertebrate body depends on anabolic and catabolic activities that are closely linked the inside and outside of the cell. Lipid metabolism plays an essential role in these metabolic activities. Although a large amount of evidence shows that normal lipid metabolism guarantees the conventional physiological activities of organs in the vertebrate body and that abnormal lipid metabolism plays an important role in the occurrence and deterioration of cardiovascular-related diseases, such as obesity, atherosclerosis, and type II diabetes, little is known about the role of lipid metabolism in cartilage and its diseases. This review aims to summarize the latest advances about the function of lipid metabolism in cartilage and its diseases including osteoarthritis, rheumatoid arthritis, and cartilage tumors. With the gradual in-depth understanding of lipid metabolism in cartilage, treatment methods could be explored to focus on this metabolic process in various cartilage diseases.


Assuntos
Artrite Reumatoide/metabolismo , Cartilagem Articular/metabolismo , Condrócitos/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Metabolismo dos Lipídeos , Osteoartrite/metabolismo , Artrite Reumatoide/patologia , Cartilagem Articular/patologia , Condrócitos/patologia , Diabetes Mellitus Tipo 2/patologia , Humanos , Osteoartrite/patologia
20.
Aging (Albany NY) ; 13(3): 4647-4662, 2021 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-33526719

RESUMO

Exosomes are major mediators of cell-to-cell communication, and are involved in many physiological and pathological processes. Recently, the roles of exosomes in osteoarthritis (OA) and their therapeutic potential have received increasing attention. Exosomes derived from vascular endothelial cells have been confirmed to participate in the occurrence and development of numerous diseases; however, their effects in OA have not been reported. Here, we demonstrated the roles of exosomes secreted by vascular endothelial cells in the development of OA. Through in vivo and in vitro experiments, we demonstrated that exosomes derived from vascular endothelial cells decreased the ability of chondrocytes to resist oxidative stress by inhibiting autophagy and p21 expression, thereby increasing the cellular ROS content and inducing apoptosis. These findings indicate that exosomes derived from vascular endothelial cells promote the progression of OA, thus, providing new ideas for the diagnosis and treatment of OA.


Assuntos
Apoptose/fisiologia , Condrócitos , Células Endoteliais/metabolismo , Exossomos , Osteoartrite , Estresse Oxidativo/fisiologia , Animais , Células Cultivadas , Condrócitos/patologia , Condrócitos/fisiologia , Exossomos/química , Exossomos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Osteoartrite/patologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...