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1.
Int J Nanomedicine ; 15: 3771-3790, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32547027

RESUMO

Introduction: Rapamycin has been considered as a potential treatment for osteoarthritis (OA). Drug carriers fabricated from liposomes can prolong the effects of drugs and reduce side effects of drugs. Low-intensity pulsed ultrasound (LIPUS) has been found to possess anti-OA effects. Materials and Methods: The anti-osteoarthritic effects of liposome-encapsulated rapamycin (L-rapa) combined with LIPUS were examined by culture of normal and OA chondrocytes in alginate beads and further validated in OA prone Dunkin-Hartley guinea pigs. Results: L-rapa with LIPUS largely up-regulated aggrecan and type II collagen mRNA in human OA chondrocytes (HOACs). L-rapa with LIPUS caused significant enhancement in proteoglycan and type II collagen production in HOACs. Large decreases in both MMP-13 and IL-6 proteins were found in the HOACs exposed to L-rapa with LIPUS. Intra-articular injection of 40 µL L-rapa at both 5 µM and 50 µM twice a week combined with LIPUS thrice a week for 8 weeks significantly increased GAGs and type II collagen in the cartilage of knee. Results on OARSI score showed that intra-articular injection of 5 µM L-rapa with LIPUS displayed the greatest anti-OA effects. Immunohistochemistry revealed that L-rapa with or without LIPUS predominantly reduced MMP-13 in vivo. The values of complete blood count and serum biochemical examinations remained in the normal ranges after the injections with or without LIPUS. These data indicated that intra-articular injection of L-rapa collaborated with LIPUS is not only effective against OA but a safe OA therapy. Conclusion: Taken together, L-rapa combined with LIPUS possessed the most consistently and effectively anabolic and anti-catabolic effects in HOACs and the spontaneous OA guinea pigs. This study evidently revealed that liposome-encapsulation collaborated with LIPUS is able to reduce the effective dose and administration frequency of rapamycin and further stably reinforce its therapeutic actions against OA.


Assuntos
Osteoartrite/terapia , Sirolimo/uso terapêutico , Ondas Ultrassônicas , Animais , Peso Corporal/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/efeitos da radiação , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Condrócitos/efeitos da radiação , Colágeno Tipo II/metabolismo , Liberação Controlada de Fármacos , Cobaias , Humanos , Injeções Intra-Articulares , Interleucina-6/metabolismo , Lipossomos/ultraestrutura , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Pessoa de Meia-Idade , Osteoartrite/sangue , Osteoartrite/patologia , Proteoglicanas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sirolimo/administração & dosagem , Sirolimo/farmacologia
2.
Chem Biol Interact ; 325: 109088, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32360554

RESUMO

Osteoarthritis (OA) is one of the most common degenerative joint diseases in aging people. The activation of chondrocytes and their dysregulation are closely related to the pathogenesis of OA. GPR55 is an unique orphan G-receptor which binds to cannabinoids. In this study, we explored the role of GPR55 in advanced glycation end productions (AGEs)- induced chondrocytes activation in cultured cells. We showed that AGEs dose dependently induced GPR55 expression in ATDC5 chondrocytes. The blockage of GPR55 by its newly discovered antagonist-CID16020046 mitigated AGEs- induced increase in cellular ROS and decrease in antioxidant NRF2. Moreover, CID16020046 showed a dose-response suppressive effect on AGEs- induced expression of the major inflammatory mediators, including COX-2 and iNOS, and the production of NO and PGE2. CID16020046 also dose responsively inhibited AGEs- induced key effectors of cartilage degradation such as MMP-3 and MMP-13. In consequence, CID16020046 showed robust inhibition on AGEs- induced type II collagen degradation. Mechanistically, our data demonstrated that CID16020046 mediated GPR55 blockage ameliorated AGEs- induced NF-κB activation as revealed by its inhibition on IκBα, nuclear p65 translocation and NF-κB promoter activity. Collectively, our study demonstrates that GPR55 signaling mediates AGEs- induced chondrocyte activation, and the targeted blockage of GPR55 pathway could be therapeutic choice in the treatment of osteoarthritis.


Assuntos
Compostos Azabicíclicos/farmacologia , Benzoatos/farmacologia , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Produtos Finais de Glicação Avançada/farmacologia , Receptores de Canabinoides/metabolismo , Linhagem Celular , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Ciclo-Oxigenase 2/genética , Dinoprostona/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Humanos , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 3 da Matriz/genética , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/biossíntese , Estresse Oxidativo/efeitos dos fármacos , Proteólise/efeitos dos fármacos
3.
Chem Biol Interact ; 326: 109136, 2020 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-32417162

RESUMO

Osteoarthritis (OA) is a common degenerative joint disease that is closely associated with inflammation. Stachydrine (STA) is a bioactive alkaloid with anti-inflammatory activity. However, the role of STA in OA remains unknown. This study aimed to explore the effects of STA on OA chondrocytes in the presence of IL-1ß. Primary human OA chondrocytes were pretreated with various concentrations of STA for 2 h and then stimulated with IL-1ß for 24 h. Inflammatory mediators and cytokines including NO, PGE2, TNF-α and IL-6 in chondrocytes were detected to reflect inflammation status. Production of extracellular matrix (ECM) degrading enzymes including MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5 in chondrocytes was measured using ELISA. The expression levels of iNOS, COX-2, p65, p-p65, p-IκBα, and IκBα were detected by Western blot analysis. Our results showed that STA significantly suppressed IL-1ß-induced inflammation with decreased levels of inflammatory mediators and cytokines including NO, PGE2, iNOS, COX-2, TNF-α and IL-6. Treatment with STA suppressed the production of ECM degrading enzymes including MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5 in IL-1ß-induced chondrocytes. Furthermore, STA blocked the IL-1ß-mediated potentiation of NF-κB pathway in chondrocytes. In conclusion, these findings demonstrated that STA protected chondrocytes from IL-1ß-induced inflammation through the NF-κB signaling pathway.


Assuntos
Condrócitos/efeitos dos fármacos , Inflamação/tratamento farmacológico , Interleucina-1beta/metabolismo , NF-kappa B/metabolismo , Osteoartrite/tratamento farmacológico , Prolina/análogos & derivados , Transdução de Sinais/efeitos dos fármacos , Anti-Inflamatórios/farmacologia , Linhagem Celular , Condrócitos/metabolismo , Humanos , Inflamação/metabolismo , Osteoartrite/metabolismo , Prolina/farmacologia
4.
Gene ; 751: 144764, 2020 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-32428694

RESUMO

Bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (Exos) have anti-inflammatory and anti-apoptotic functions. miRNA-210 has also been confirmed to play a role in inhibiting proinflammatory cytokines. Herein, we aimed to explore the effects of Exos derived from miRNA-210-overexpressing BMSCs (BMSCs-210-Exos) and the mechanisms by which they provide protection to chondrocytes from lipopolysaccharide (LPS)-induced injury. BMSCs were transfected with or without miRNA-210. Exos substantially improved the proliferation of chondrocytes and inhibited LPS-induced cell apoptosis. Furthermore, BMSCs-210-Exos promoted the proliferation of chondrocytes and prevented LPS-induced cell apoptosis better than BMSCs-Exos not overexpressing miRNA-210. In addition, tumor necrosis factor receptor superfamily member 21 (Tnfrsf21) expression was inhibited and the NF-κB pathway was attenuated by both BMSCs-Exos and BMSCs-210-Exos during LPS-induced chondrocyte injury. Collectively, these results suggest that BMSCs-210-Exos enhance the protection of chondrocytes from LPS-induced injury via the NF-κB pathway.


Assuntos
Condrócitos/metabolismo , Exossomos/fisiologia , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/metabolismo , NF-kappa B/metabolismo , Animais , Apoptose , Proliferação de Células , Células Cultivadas , Condrócitos/citologia , Condrócitos/efeitos dos fármacos , Exossomos/ultraestrutura , Lipopolissacarídeos/toxicidade , Células-Tronco Mesenquimais/ultraestrutura , Camundongos , Transdução de Sinais
5.
Zhongguo Zhong Yao Za Zhi ; 45(5): 1097-1104, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-32237452

RESUMO

Uniform design-comprehensive scoring method was used to investigate the effects of ethanol dosage, ethanol concentration and extraction time, based on the evaluation index from transfer rates of epimedin A, epimedin B, epimedin C, icariin and baohuoside Ⅰ, which are the main active components in Epimedii Folium. Furthermore, the optimum conditions for the ethanol extraction process were determined by multiple linear stepwise regression and empirical test. Then, the ethanol extract of Epimedii Folium prepared according to the optimized technological conditions was used to intervene the injury model of chondrocyte induced by interleukin-1 beta(IL-1ß). Annexin V-FITC/PI staining flow cytometry was used to detect the apoptotic rate of chondrocyte and analyze the effect of ethanol extract of Epimedii Folium on chondrocyte injury model. The optimum conditions of ethanol extraction were as follows. Crude powder of Epimedii Folium was added with 18 times of 70% ethanol solution, and extracted for twice in the refluxing process, for 60 minutes each time. Under the conditions, the extraction rates of the above five active components were 94.21%, 94.76%, 93.85%, 96.17% and 96.85%, respectively. The optimized ethanol extraction process of Epimedii Folium was reasonable, feasible and reproducible. This ethanol extract could significantly reduce the early apoptotic rate, late apoptotic and necrotic rate, total apoptotic rate(P<0.05 or P<0.01) of chondrocyte injury model induced by IL-1ß, suggesting that the ethanol extract of Epimedii Folium can inhibit the apoptosis of chondrocytes induced by IL-1ß to a certain extent, which lays a foundation for further study on its prevention and treatment of osteoarthritis.


Assuntos
Condrócitos/efeitos dos fármacos , Epimedium/química , Etanol , Extratos Vegetais/farmacologia , Folhas de Planta/química , Humanos , Interleucina-1beta
6.
Yonsei Med J ; 61(4): 331-340, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32233176

RESUMO

PURPOSE: Osteoarthritis (OA) of the temporomandibular joint (TMJ) elicits cartilage and subchondral bone defects. Growth hormone (GH) promotes chondrocyte growth. The aim of this study was to evaluate the efficacy of intra-articular injections of GH to treat TMJ-OA. MATERIALS AND METHODS: Monosodium iodoacetate (MIA) was used to induce OA in the TMJs of rats. After confirming the induction of OA, recombinant human GH was injected into the articular cavities of rats. Concentrations of GH and IGF-1 were measured in the blood and synovial fluid, and OA grades of cartilage and subchondral bone degradation were recorded by histological examination and micro-computed tomography. RESULTS: MIA-induced OA in the rat TMJ upregulated insulin-like growth factor-1 (IGF-1) rather than GH levels. GH and IGF-1 concentrations were increased after local injection of GH, compared with controls. Locally injected GH lowered osteoarthritic scores in the cartilage and subchondral bone of the TMJ. CONCLUSION: Intra-articular injection of GH improved OA scores in rat TMJs in both cartilage and subchondral bone of the condyles without affecting condylar bone growth. These results suggest that intra-articular injection of human GH could be a suitable treatment option for TMJ-OA patients in the future.


Assuntos
Condrócitos/efeitos dos fármacos , Hormônio do Crescimento/administração & dosagem , Fator de Crescimento Insulin-Like I/administração & dosagem , Osteoartrite/tratamento farmacológico , Articulação Temporomandibular/efeitos dos fármacos , Idoso , Animais , Cartilagem Articular/diagnóstico por imagem , Cartilagem Articular/patologia , Hormônio do Crescimento/efeitos adversos , Hormônio do Crescimento Humano , Humanos , Injeções Intra-Articulares , Masculino , Osteoartrite/diagnóstico por imagem , Osteoartrite/patologia , Ratos , Líquido Sinovial , Articulação Temporomandibular/diagnóstico por imagem , Articulação Temporomandibular/fisiopatologia , Microtomografia por Raio-X
7.
Ann Rheum Dis ; 79(5): 635-645, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32156705

RESUMO

OBJECTIVES: In this study, we aim to determine the effect of metformin on osteoarthritis (OA) development and progression. METHODS: Destabilisation of the medial meniscus (DMM) surgery was performed in 10-week-old wild type and AMP-activated protein kinase (AMPK)α1 knockout (KO) mice. Metformin (4 mg/day in drinking water) was given, commencing either 2 weeks before or 2 weeks after DMM surgery. Mice were sacrificed 6 and 12 weeks after DMM surgery. OA phenotype was analysed by micro-computerised tomography (µCT), histology and pain-related behaviour tests. AMPKα1 (catalytic alpha subunit of AMPK) expression was examined by immunohistochemistry and immunofluorescence analyses. The OA phenotype was also determined by µCT and MRI in non-human primates. RESULTS: Metformin upregulated phosphorylated and total AMPK expression in articular cartilage tissue. Mild and more severe cartilage degeneration was observed at 6 and 12 weeks after DMM surgery, evidenced by markedly increased Osteoarthritis Research Society International scores, as well as reduced cartilage areas. The administration of metformin, commencing either before or after DMM surgery, caused significant reduction in cartilage degradation. Prominent synovial hyperplasia and osteophyte formation were observed at both 6 and 12 weeks after DMM surgery; these were significantly inhibited by treatment with metformin either before or after DMM surgery. The protective effects of metformin on OA development were not observed in AMPKα1 KO mice, suggesting that the chondroprotective effect of metformin is mediated by AMPK signalling. In addition, we demonstrated that treatment with metformin could also protect from OA progression in a partial medial meniscectomy animal model in non-human primates. CONCLUSIONS: The present study suggests that metformin, administered shortly after joint injury, can limit OA development and progression in injury-induced OA animal models.


Assuntos
Proteínas Quinases Ativadas por AMP/genética , Cartilagem Articular/efeitos dos fármacos , Metformina/farmacologia , Osteoartrite/tratamento farmacológico , Regulação para Cima/genética , Animais , Cartilagem Articular/patologia , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Modelos Animais de Doenças , Progressão da Doença , Regulação da Expressão Gênica , Humanos , Hipoglicemiantes/farmacologia , Meniscos Tibiais/patologia , Meniscos Tibiais/cirurgia , Camundongos , Camundongos Knockout , Camundongos Obesos , Osteoartrite/patologia , Distribuição Aleatória , Sensibilidade e Especificidade , Transdução de Sinais/genética
8.
PLoS One ; 15(3): e0230228, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32163510

RESUMO

This study was designed to evaluate the anti-inflammatory effects of a curcumin treatment on the knee of rats with induced osteoarthritis. Fifteen adult rats were used and divided in three groups: the osteoarthritis group (OAG), control group (CG-without induction of osteoarthritis), and curcumin-treated osteoarthritis group (COAG). Osteoarthritis was induced in the right knee of rats in the OAG and COAG by administering an intra-articular injection of 1 mg of zymosan. Fourteen days after induction, 50 mg/kg curcumin was administered by gavage daily for 60 days to the COAG. After the treatment period, rats from all groups were euthanized. Medial femoral condyles were collected for light microscopy and immunohistochemical staining. The expression of SOX-5, IHH, MMP-8, MMP-13, and collagen 2 (Col2) was analyzed. The COAG exhibited an increase in the number of chondrocytes in the surface and middle layers compared with that of the OAG and CG, respectively. The COAG also showed a decrease in the thicknesses of the middle and deep layers compared with those of the OAG, and an increase in Col2 expression was observed in all articular layers (surface, middle, and deep) in the COAG compared with that in the OAG. SOX-5 expression was increased in the surface and deep layers of the COAG compared with those in the OAG and CG. Based on the results of this study, the curcumin treatment appeared to exert a protective effect on cartilage, as it did not result in an increase in cartilage thickness or in MMP-8 and MMP-13 expression but led to increased IHH, Col2, and SOX-5 expression and the number of chondrocytes.


Assuntos
Cartilagem Articular/efeitos dos fármacos , Curcumina/farmacologia , Articulação do Joelho/efeitos dos fármacos , Osteoartrite/tratamento farmacológico , Animais , Cartilagem Articular/metabolismo , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Colágeno Tipo II/metabolismo , Injeções Intra-Articulares/métodos , Articulação do Joelho/metabolismo , Masculino , Osteoartrite/metabolismo , Ratos , Ratos Wistar
9.
Toxicon ; 176: 34-43, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32103793

RESUMO

T-2 toxin is considered an unavoidable pollutant, which contaminates food crops and stockpiled cereals, impairing the health of humans and animals due to its multi-organ toxicity. Studies have shown that T-2 toxin can cause articular cartilage damage; however, the underlying molecular mechanism is still unclear. Here, we investigated the possible mechanism of the following inhibitors of apoptosis proteins (IAPs) family members: NAIP, cIAP1, cIAP2, XIAP, and Survivin, and their involvement in T-2 toxin-induced mouse chondrocyte damage. In this study, mouse articular chondrocytes were isolated and cultured in vitro, and the chondrocytes were then treated with 0, 5, 10, and 20 ng/mL T-2 toxin. Firstly, the toxic effect of T-2 toxin on chondrocytes was determined. CCK-8 assay results showed that T-2 toxin induced a dose-dependent inhibition of chondrocyte viability. Transmission electron microscopy demonstrated that T-2 toxin caused morphological changes in chondrocyte endoplasmic reticulum and an increase in mitochondrial swelling. In addition, Annexin-V-FITC/PI staining and caspase 3 protein expression showed that T-2 toxin induced an increase in the apoptotic rate of chondrocytes. Secondly, it was found that T-2 toxin cause decreased expression of cellular and secreted Collagen II. Finally, we examined the expression of NAIP, cIAP1, cIAP2, XIAP, and Survivin in chondrocytes in the presence of T-2 toxin and their relationship with decreased Collagen II. The decrease in Collagen II was negatively correlated with the expression of cIAP1, cIAP2 and positively correlated with NAIP and Survivin mRNA level. Survivin mRNA level had a positive correlation with Collagen II as shown by partial correlation analysis. This study revealed the new role of IAPs in chondrocyte injury and provides new insights and clues into the mechanism of T-2 toxin-induced chondrocyte damage.


Assuntos
Condrócitos/efeitos dos fármacos , Toxina T-2/toxicidade , Animais , Proteína 3 com Repetições IAP de Baculovírus/metabolismo , Cartilagem Articular/metabolismo , Caspase 3/metabolismo , Colágeno/metabolismo , Humanos , Camundongos
10.
Int J Nanomedicine ; 15: 1173-1186, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32110015

RESUMO

Background: The facile preparation of oxygen-generating microparticles (M) consisting of Polycaprolactone (PCL), Pluronic F-127, and calcium peroxide (CPO) (PCL-F-CPO-M) fabricated through an electrospraying process is disclosed. The biological study confirmed the positive impact from the oxygen-generating microparticles on the cell growth with high viability. The presented technology could work as a prominent tool for various tissue engineering and biomedical applications. Methods: The oxygen-generated microparticles fabricated through electrospraying processes were thoroughly characterization through various methods such as X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) analysis, and scanning electron microscopy (SEM)/SEM-Energy Dispersive Spectroscopy (EDS) analysis. Results: The analyses confirmed the presence of the various components and the porous structure of the microparticles. Spherical shape with spongy characteristic microparticles were obtained with negative charge surface (ζ = -16.9) and a size of 17.00 ± 0.34 µm. Furthermore, the biological study performed on rat chondrocytes demonstrated good cell viability and the positive impact of increasing the amount of CPO in the PCL-F-CPO-M. Conclusion: This technological platform could work as an important tool for tissue engineering due to the ability of the microparticles to release oxygen in a sustained manner for up to 7 days with high cell viability.


Assuntos
Oxigênio/farmacocinética , Animais , Materiais Biocompatíveis/química , Técnicas de Cultura de Células , Proliferação de Células , Sobrevivência Celular/efeitos dos fármacos , Condrócitos/efeitos dos fármacos , Técnicas Eletroquímicas , Oxigênio/química , Peróxidos/química , Poloxâmero/química , Poliésteres/química , Porosidade , Ratos Wistar , Espectrometria por Raios X , Espectroscopia de Infravermelho com Transformada de Fourier , Engenharia Tecidual/métodos , Difração de Raios X
11.
BMC Complement Med Ther ; 20(1): 23, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-32020892

RESUMO

BACKGROUND: Chondrocyte apoptosis activated by the mitochondrial dependent pathway serves a crucial role in cartilage degeneration of osteoarthritis (OA). In the present study, the protective effects of CMCS against sodium nitroprusside (SNP)-induced chondrocyte apoptosis were evaluated and the underlying molecular mechanisms were elucidated. METHODS: Chondrocytes were isolated from articular cartilage of SD rats and identified by type II collagen immunohistochemistry. The chondrocytes stimulated with or without SNP to induce apoptosis, were treated by CMCS for various concentrations. The cell viability were determined by MTT and LDH assays. Cell apoptotic ratio was determined by Annexin V-FITC/PI staining. Mitochondrial membrane potential (ΔΨm) was detected by using Rhodamine123 (Rho123) staining. To understand the mechanism, the mRNA expression levels of Bcl-2, Bax, cytochrome c (Cyt c) and cleaved caspase-3 were detected by real-time PCR and western blot analysis, respectively. RESULTS: It was shown using the MTT and LDH assays that CMCS protected the viability of chondrocyte against SNP damage. Annexin V-FITC/PI and Rho123 staining showed that CMCS not only inhibited the cell apoptosis but also restored the reduction of the ΔΨm in chondrocytes. In SNP-induced chondrocytes, CMCS down-regulated the expression of Bax, Cyt c and cleaved caspase-3 but upregulated the expression of Bcl-2, as shown by real-time PCR and western blot. CONCLUSIONS: Taken together, these results indicated that CMCS has the protective effect on chondrocytes against SNP-induced apoptosis, at least partly, via inhibiting the mitochondrial dependent apoptotic pathway. Thus, CMCS may be potentially used as a biological agent for prevention and treatment of OA.


Assuntos
Apoptose/efeitos dos fármacos , Quitosana/farmacologia , Condrócitos/efeitos dos fármacos , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Animais , Células Cultivadas , Quitosana/química , Condrócitos/citologia , Óxido Nítrico , Nitroprussiato , Ratos , Ratos Sprague-Dawley
12.
J Bone Miner Metab ; 38(3): 346-356, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-31894489

RESUMO

INTRODUCTION: Estrogen receptor α (ERα) plays important roles in the etiology of osteoarthritis (OA), in which cartilage degradation and cellular inflammation are involved. MiR-203 is reported to direct target ERα, but its roles in chondrocytes remain uncovered. METHODS: In this study, ELISA showed that the level of estrogen hormone in the serum of postmenopausal OA patients was significantly lower than the one in patients without OA. RT-PCR revealed that the expression level of miR-203 was significantly up-regulated in the OA patients. Furthermore, western blotting demonstrated the lower expression levels of aggrecan, Col2A1, and ERα in the isolated articular cartilage tissues of OA patients. To decipher the association between ERα and miR-203 in the pathogenesis of OA, IL-1ß stimulated cultured chondrocyte cell model was established to measure the cell viability, cellular inflammation, cell injury, as well as cartilage degradation with miR-203 inhibitor and ERα. RESULTS: The results showed that IL-1ß stimulation induced the expression of miR-203, which promoted cellular inflammation and cell injury, and caused down-regulation of aggrecan and Col2A1. Luciferase assay indicated the direct binding between miR-203 and ERα, and ERα-specific SiRNA inversed the protective role of miR-203 inhibitor in the progression of OA in the cell system. CONCLUSIONS: MiR-203 is critical in the onset and progression of OA, at least in part, caused by estrogen deficiency and ERα instability in OA patients, providing a novel therapeutic target for the treatment of OA.


Assuntos
Condrócitos/metabolismo , Receptor alfa de Estrogênio/metabolismo , Interleucina-1beta/farmacologia , MicroRNAs/metabolismo , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Feminino , Humanos , Inflamação/genética , Inflamação/patologia , MicroRNAs/genética , Pessoa de Meia-Idade , Osteoartrite/genética , Osteoartrite/patologia
13.
Life Sci ; 243: 117244, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31891721

RESUMO

AIMS: Endplate chondrocyte apoptosis is an important contributor to the pathogenesis of cartilaginous endplate (CEP) degeneration that leads to the initiation and development of intervertebral disc degeneration (IDD). In this study, we hypothesized that Parkin-mediated mitophagy and nuclear factor E2-related factor 2 (Nrf2)-mediated antioxidant system played an important role in endplate chondrocyte survival under pathological conditions. MATERIALS AND METHODS: Human endplate chondrocytes were stimulated with H2O2 to mimic pathological conditions. Western blotting, immunofluorescence staining, and flow cytometry were applied to detect the indicators related to mitochondrial dynamics, mitophagy, Nrf2 signaling, and apoptosis. The puncture-induced rat models were established to evaluate the changes in vivo. KEY FINDINGS: Our results showed that H2O2 induced oxidative stress, mitochondrial dysfunction, and apoptosis in endplate chondrocytes. These H2O2-induced detrimental effects were inhibited by pretreatment with the mitochondria-targeted antioxidant Mito-TEMPO. In addition, mitochondrial dynamics, Parkin-mediated elimination of dysfunctional mitochondria, and Nrf2-mediated antioxidant system were promoted by H2O2. Knockdown of Parkin or Nrf2 increased H2O2-induced detrimental effects. Moreover, upregulation of Parkin and Nrf2 by polydatin protected endplate chondrocytes against H2O2-induced mitochondrial dysfunction, oxidative stress, and apoptosis. Finally, puncture-induced rat models showed that polydatin exerted a protective effect on CEP and disc degeneration. SIGNIFICANCE: Targeting Parkin and Nrf2 to improve mitochondrial homeostasis, redox balance and endplate chondrocyte survival may represent a potential therapeutic strategy for preventing IDD.


Assuntos
Apoptose/fisiologia , Condrócitos/patologia , Disco Intervertebral/patologia , Mitofagia/fisiologia , Fator 2 Relacionado a NF-E2/fisiologia , Estresse Oxidativo/fisiologia , Ubiquitina-Proteína Ligases/fisiologia , Idoso , Animais , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Feminino , Técnicas de Silenciamento de Genes , Humanos , Peróxido de Hidrogênio/farmacologia , Disco Intervertebral/efeitos dos fármacos , Disco Intervertebral/metabolismo , Masculino , Pessoa de Meia-Idade , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo
14.
Gene ; 732: 144339, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31927008

RESUMO

OBJECTIVE: Previous studies have shown that follistatin-like protein 1 (FSTL1) is elevated in the synovial fluid of osteoarthritis and is associated with disease activity. The experiment was performed to stuy the effect and mechanism of FSTL1 on chondrocyte apoptosis in osteoarthritis. DESIGN: After the isolation of human normal and osteoarthritis (OA) chondrocytes, the expression of FSTL1 was detected by Q-PCR and western blot analyses. Chondrocytes were pre-transfected with FSTL1 overexpression plasmids then treated with SNP, and chondrocyte viability and apoptosis levels were detected by MTS and flow cytometry, respectively. Cartilage matrix gene expression was measured by Q-PCR and signal pathway-related proteins were assessed by western blot. RESULTS: The expression of FSTL1 in OA chondrocytes was markedly up-regulated compared with normal human chondrocytes (P < 0.05). The apoptosis rate of chondrocytes in the FSTL1 overexpression groups was highly elevated in the comparison with the negative control groups (P < 0.05). Additionally, FSTL1 potentiated protein abundances of MMP1, MMP3, MMP-9, and Bax as well as reduced Coll2a1 and Aggrecan and Bcl-2 expression. Furthermore, western blot results showed that the SAPK/JNK/Caspase3 signal pathway was significantly activated and the Ac-DEVD-FMK impaired FSTL1 induced chondrocyte apoptosis. CONCLUSION: FSTL1 promoted SNP-induced chondrocytes apoptosis by activating the SAPK/JNK/Caspase3 signal pathway.


Assuntos
Apoptose/fisiologia , Caspase 3/metabolismo , Condrócitos/citologia , Proteínas Relacionadas à Folistatina/fisiologia , MAP Quinase Quinase 4/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Óxido Nítrico/fisiologia , Transdução de Sinais/fisiologia , Idoso , Apoptose/efeitos dos fármacos , Estudos de Casos e Controles , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Proteínas Relacionadas à Folistatina/genética , Humanos , Pessoa de Meia-Idade , Osteoartrite/enzimologia , Osteoartrite/metabolismo , Osteoartrite/patologia , RNA Mensageiro/metabolismo
15.
Gene ; 730: 144322, 2020 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-31899303

RESUMO

AIM: This study aimed to investigate the effect of 1,25-dihydroxy-vitamin D3 (1,25(OH)2D3) on primary chondrocytes cultured from patients with osteoarthritis (OA). METHOD: Primary chondrocytes isolated from the tibial plateau of female OA patients were characterized by immunocytochemistry analysis. Using Cell Counting Kit-8 (CCK-8), cell viability was measured to select suitable 1,25(OH)2D3 concentrations for treating chondrocytes. RNA-sequencing was performed on primary chondrocytes treated with or without 1,25(OH)2D3. Differentially expressed genes (DEGs) as well as gene ontology (GO)-biological process (BP) and pathways affected by 1,25(OH)2D3 were identified. Protein-protein interaction (PPI) network was constructed, and the hub nodes in the PPI network were identified. qRT-PCR was conducted to confirm the expression levels of six DEGs. RESULTS: Positive collagen II staining confirmed the successful isolation of primary chondrocytes. CCK-8 assay showed maximal primary chondrocyte survival rate when treated with 10-5 µmol/L of 1,25(OH)2D3 for 72 h. RNA-sequencing results identified a total of 1036 DEGs, including 593 upregulated and 443 downregulated genes from 1,25(OH)2D3 treated and untreated cells. Further functional enrichment analyses showed the association of these DEGs with GO-BP terms such as response to the stimulus, cell proliferation, angiogenesis, and regulation of cell motility, and KEGG pathways, including TNF signaling pathway, IL-17 signaling pathway, cytokine-cytokine receptor interaction, and NF-kappa B signaling pathway. PPI network identified UBC, FOS, IFIT1, CDK1, and ISG15 as the hub nodes in the network. qRT-PCR results were in alignment with the results of RNA-sequencing. CONCLUSION: Our study might provide a theoretical basis for the use of vitamin D in treating OA.


Assuntos
Condrócitos/efeitos dos fármacos , Vitamina D/análogos & derivados , Idoso , Feminino , Ontologia Genética , Humanos , Pessoa de Meia-Idade , Osteoartrite/metabolismo , Cultura Primária de Células , Análise de Sequência de RNA , Transdução de Sinais/efeitos dos fármacos , Transcriptoma/genética , Vitamina D/metabolismo , Vitamina D/farmacologia , Vitamina D/fisiologia , Vitamina D/uso terapêutico , Vitaminas/farmacologia
16.
Clin Exp Rheumatol ; 38(1): 129-135, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31172921

RESUMO

OBJECTIVES: Prolonged use of glucocorticoids (GCs) for treatment of inflammatory and autoimmune conditions may have several negative side effects, such as impaired bone growth which has been linked to increased apoptosis in growth plate chondrocytes. It has recently been shown that humanin, a small mitochondrial derived peptide, rescues growth plate chondrocytes from GC-induced apoptosis. Our aim was to study if a synthetic analogue of humanin, [Gly14]-HNG (HNG), can be safely used to prevent GC-induced toxicity in growth plate chondrocytes without interfering with the desired anti-inflammatory effect in an in vivo model of arthritis. METHODS: Arthritis was induced in DBA/1 mice by collagen type II in complete Freund's adjuvant and the animals were treated with Dexamethasone (Dexa) (0.25 mg/kg/day) with or without HNG (100 µg/kg/day) for 14 days. The animals were observed daily for the presence of arthritis including signs of erythema and swelling of the joints. The paws were scored based on the severity of the swelling. After termination, histological scoring was performed of all paws. Chondrocyte apoptosis and proliferation were analysed by TUNEL assay and PCNA staining, respectively. RESULTS: We found that HNG treatment in combination with Dexa protected from Dexa-induced chondrocyte apoptosis in both articular and growth plate cartilage. Furthermore, based on clinical and histology scoring analyses, HNG did not interfere with the desired anti-inflammatory effect of Dexa. CONCLUSIONS: Our results suggest that the combination of HNG and GCs may provide a new treatment strategy in conditions of chronic inflammation, which could potentially prevent bone growth impairment.


Assuntos
Anti-Inflamatórios , Apoptose , Artrite Experimental , Dexametasona , Peptídeos e Proteínas de Sinalização Intracelular , Animais , Anti-Inflamatórios/farmacologia , Apoptose/efeitos dos fármacos , Artrite Experimental/tratamento farmacológico , Condrócitos/efeitos dos fármacos , Dexametasona/farmacologia , Peptídeos e Proteínas de Sinalização Intracelular/farmacologia , Camundongos , Camundongos Endogâmicos DBA
17.
Life Sci ; 240: 116857, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31521691

RESUMO

AIMS: Daphnetin (DAP) is a traditional Chinese drug usually used to treat cardiovascular diseases. Studies have confirmed the anti-inflammatory, antioxidant, anti-bacterial and insecticidal, anti-tumor and neuro-protective effects of DAP. However, its anti-arthritic potential remains unexplored. The aim of this study is to investigate the in vitro and in vivo chondroprotective effects of DAP. MAIN METHODS: The effect of DAP on primary rabbit chondrocytes was examined using recombinant human IL-1ß for 24 h. For the in vivo studies, rabbits were randomly divided into groups: a normal control group and osteoarthritis (OA) groups. The OA groups received three different doses of DAP for 4 or 8 weeks. The anti-arthritic effect of DAP was assessed using histopathological examinations, qRT-PCR, western blotting and immunohistochemical analysis. KEY FINDINGS: Both in vitro and in vivo results indicate that DAP exerts a protective effect against IL-1ß in chondrocytes. In vitro, DAP inhibits the expression of IL-6, IL-12, MMP-3, MMP-9 and MMP-13, induced by IL-1ß in rabbit chondrocytes, and stimulates the production of IL-10. The inhibitory effect of DAP on the MMPs is partially regulated by the inhibition of the PI3K/AKT, MAPK and NF-κB signaling pathways. The effect of DAP on OA may be attributed to the suppression of inflammatory factor secretion, chondrocyte apoptosis observed by the decrease in pro-apoptotic Caspase-3 and BAX, and the activation of anti-apoptotic BCL-2. SIGNIFICANCE: DAP has a broad range of prospects in the treatment of OA, which provides a novel therapeutic strategy for OA.


Assuntos
Antirreumáticos/uso terapêutico , Condrócitos/efeitos dos fármacos , Osteoartrite/tratamento farmacológico , Substâncias Protetoras/uso terapêutico , Umbeliferonas/uso terapêutico , Animais , Antirreumáticos/efeitos adversos , Apoptose/efeitos dos fármacos , Cartilagem Articular/efeitos dos fármacos , Cartilagem Articular/patologia , Sobrevivência Celular , Condrócitos/patologia , Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Relação Dose-Resposta a Droga , Interleucina-1beta/farmacologia , Masculino , Osteoartrite/patologia , Cultura Primária de Células , Coelhos , Transdução de Sinais/efeitos dos fármacos , Umbeliferonas/efeitos adversos
18.
Biosci Biotechnol Biochem ; 84(2): 402-410, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31642732

RESUMO

This study was conducted to evaluate the protective effects of chamazulene against IL-1ß-induced rat primary chondrocytes and complete Freund's adjuvant (CFA)-induced osteoarthritic inflammation in rats. Oxidative stress markers, pro-inflammatory cytokines, and regulatory proteins were measured. Chamazulene significantly reverted (p < 0.05) the levels of lipid peroxidation and enhanced the activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) enzymes against IL-1ß and CFA-induced oxidative stress. The levels of TNF-α and IL-6 were reduced (p < 0.05) in chamazulene treatment against IL-1ß and CFA-induced inflammation. Western blot analysis results on the expressions of MMP-3, MMP-9, p65 NF-kß, iNOS, and COX-2 showed chamazulene was able to protect the chondrocytes against IL-1ß-induced osteoarthritic inflammation. Histopathology of rat hind ankle showed chamazulene significantly protected against CFA-induced osteoarthritic inflammation. Therefore, chamazulene can be recommended as a therapeutic agent for clinical trials against osteoarthritic inflammation.


Assuntos
Azulenos/uso terapêutico , Metaloproteinases da Matriz/metabolismo , Modelos Biológicos , NF-kappa B/metabolismo , Osteoartrite/prevenção & controle , Animais , Peso Corporal/efeitos dos fármacos , Células Cultivadas , Condrócitos/efeitos dos fármacos , Técnicas In Vitro , Interleucina-1beta/farmacologia , Masculino , Osteoartrite/enzimologia , Osteoartrite/metabolismo , Osteoartrite/patologia , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
19.
World Neurosurg ; 133: e165-e172, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31476465

RESUMO

BACKGROUND: Cartilaginous endplate (CEP), a thin layer of hyaline cartilage located between the vertebral endplate and nucleus pulposus, transports the nutrient into the disc. The objective of this study was to evaluate the influence of T140 (polyphemusin II-derived peptide) on the CEP cell growth, apoptosis, and the matrix formation via the stromal cell-derived factor-1 (SDF-1)/cysteine X cysteine (CXC) receptor-4 (CXCR4) signaling pathway. METHODS: Sprague-Dawley rats were euthanized by cervical dislocation and dissected for the isolation and the appraisal of CEP cells that were extracted from the endplate in rat intervertebral discs and were then added with different concentrations of reagents (SDF-1 and T140). The effect of T140 on CEP cell proliferation and apoptosis were analyzed. The messenger RNA (mRNA) and protein expressions of CXCR4, prominin-1, proteoglycans, type II collagen, B-cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein were analyzed by reverse transcription quantitative polymerase chain reaction and Western blot analysis. RESULTS: T140 promoted the proliferation of CEP cells and inhibited the apoptosis of CEP cells. Additionally, T140 suppressed the mRNA and protein expression of CXCR4, prominin-1, and Bcl-2 associated X protein, and increased the mRNA and protein expression of proteoglycans, type II collagen, and Bcl-2. CONCLUSIONS: T140 promotes the proliferation and matrix formation and inhibits the apoptosis of CEP cells by blocking the SDF-1/CXCR4 signaling pathway in vitro, which provides a certain therapeutic effect on the degeneration of intervertebral discs.


Assuntos
Apoptose/efeitos dos fármacos , Quimiocina CXCL12/fisiologia , Condrócitos/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Cartilagem Hialina/citologia , Disco Intervertebral/citologia , Oligopeptídeos/farmacologia , Receptores CXCR4/fisiologia , Transdução de Sinais/efeitos dos fármacos , Animais , Divisão Celular/efeitos dos fármacos , Proteínas da Matriz Extracelular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Disco Intervertebral/efeitos dos fármacos , Masculino , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley
20.
Biosci Biotechnol Biochem ; 84(4): 695-702, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31809639

RESUMO

Emerging evidence has shown that microRNAs are important regulators in osteoarthritis (OA). Here, we investigated the function role of miR-455-3p in the pathogenesis of OA and the underlying molecular mechanisms. We first established the in vitro OA model using IL-1ß treated human chondrocyte cell line CHON-001. Using quantitative real time PCR, we observed the expression of miR-455-3p expression was up-regulated in the OA cartilage tissues and IL-1ß-treated chondrocytes. A series of function assays, including CCK-8 assay, flow cytometry, and ELISA assay showed that miR-455-3p contributed to IL-1ß-induced apoptosis and inflammation. Moreover, COL2A1 was confirmed as a target of miR-455-3p by luciferase reporter assay. Furthermore, COL2A1 knockdown reversed the effects of miR-455-3p inhibition, and aggravated the effects of miR-455-3p overexpression on IL-1ß-induced OA-like phenomenon. Taken together, these results revealed that miR-455-3p/COL2A1 axis might provide a novel molecular target for the treatment of OA.


Assuntos
Apoptose/genética , Condrócitos/citologia , Colágeno Tipo II/metabolismo , Inflamação/patologia , MicroRNAs/fisiologia , Osteoartrite/patologia , Regiões 3' não Traduzidas , Idoso , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Colágeno Tipo II/genética , Feminino , Técnicas de Silenciamento de Genes , Humanos , Técnicas In Vitro , Inflamação/metabolismo , Interleucina-1beta/farmacologia , Masculino , Pessoa de Meia-Idade , Osteoartrite/metabolismo
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