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1.
Biomed Pharmacother ; 138: 111537, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34311535

RESUMO

Aging of the skin is a complicated bioprocess that is affected by constant exposure to ultraviolet irradiation. The application of herbal-based anti-aging creams is still the best choice for treatment. In the present study, Citrus sinensis L. fruit peels ethanolic extract (CSPE) was formulated into lipid nanoparticles (LNPs) anti-aging cream. Eight different formulations of CSEP-LNPs were prepared and optimized using 23 full factorial designs. In vivo antiaging effect of the best formula was tested in Swiss albino mice where photo-aging was induced by exposure to UV radiation. HPLC-QToF-MS/MS metabolic profiling of CSPE led to the identification of twenty-nine metabolites. CSPE was standardized to a hesperidin content of 15.53 ± 0.152 mg% using RP-HPLC. It was suggested that the optimized formulation (F7) had (245 nm) particle size, (91.065%) EE, and (91.385%) occlusive effect with a spherical and smooth surface. The visible appearance of UV-induced photoaging in mice was significantly improved after topical application on CSPE-NLC cream for 5 weeks, levels of collagen and SOD were significantly increased in CSPE- NLC group, while levels of PGE2, COX2, JNK, MDA, and elastin was reduced. Finally, The prepared anti-aging CSPE-NLC cream represents a safe, convenient, and promising skincare cosmetic product.


Assuntos
Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Citrus sinensis , Metaloproteinase 13 da Matriz/metabolismo , Extratos Vegetais/administração & dosagem , Envelhecimento da Pele/efeitos dos fármacos , Creme para a Pele/administração & dosagem , Pele/efeitos dos fármacos , Administração Cutânea , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Citrus sinensis/química , Colágeno/metabolismo , Regulação para Baixo , Composição de Medicamentos , Feminino , Frutas , Lipídeos/química , Metaloproteinase 13 da Matriz/genética , Camundongos , Nanopartículas , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Extratos Vegetais/isolamento & purificação , Pele/enzimologia , Pele/patologia , Pele/efeitos da radiação , Creme para a Pele/química , Creme para a Pele/isolamento & purificação , Superóxido Dismutase/metabolismo , Raios Ultravioleta
2.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201564

RESUMO

Obesity increases the risk of hip osteoarthritis (OA). Recent studies have shown that adipokine extracellular nicotinamide phosphoribosyltransferase (eNAMPT or visfatin) induces the production of IL-6 and matrix metalloproteases (MMPs) in chondrocytes, suggesting it may promote articular cartilage degradation. However, neither the functional effects of extracellular visfatin on human articular cartilage tissue, nor its expression in the joint of hip OA patients of varying BMI, have been reported. Hip OA joint tissues were collected from patients undergoing joint replacement surgery. Cartilage explants were stimulated with recombinant human visfatin. Pro-inflammatory cytokines and MMPs were measured by ELISA and Luminex. Localisation of visfatin expression in cartilage tissue was determined by immunohistochemistry. Cartilage matrix degradation was determined by quantifying proteoglycan release. Expression of visfatin was elevated in the synovial tissue of hip OA patients who were obese, and was co-localised with MMP-13 in areas of cartilage damage. Visfatin promoted the degradation of hip OA cartilage proteoglycan and induced the production of pro-inflammatory cytokines (IL-6, MCP-1, CCL20, and CCL4) and MMPs. The elevated expression of visfatin in the obese hip OA joint, and its functional effects on hip cartilage tissue, suggests it plays a central role in the loss of cartilage integrity in obese patients with hip OA.


Assuntos
Cartilagem Articular/patologia , Citocinas/metabolismo , Nicotinamida Fosforribosiltransferase/metabolismo , Osteoartrite do Quadril/metabolismo , Idoso , Idoso de 80 Anos ou mais , Cartilagem Articular/metabolismo , Quimiocinas/metabolismo , Condrócitos/metabolismo , Citocinas/sangue , Articulação do Quadril/metabolismo , Articulação do Quadril/fisiopatologia , Humanos , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinases da Matriz/metabolismo , Pessoa de Meia-Idade , NAD/metabolismo , Nicotinamida Fosforribosiltransferase/sangue , Obesidade/metabolismo , Técnicas de Cultura de Órgãos , Osteoartrite do Quadril/patologia , Proteoglicanas/metabolismo
3.
J Cell Mol Med ; 25(12): 5671-5680, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-33942503

RESUMO

Iron overload is common in elderly people which is implicated in the disease progression of osteoarthritis (OA), however, how iron homeostasis is regulated during the onset and progression of OA and how it contributes to the pathological transition of articular chondrocytes remain unknown. In the present study, we developed an in vitro approach to investigate the roles of iron homeostasis and iron overload mediated oxidative stress in chondrocytes under an inflammatory environment. We found that pro-inflammatory cytokines could disrupt chondrocytes iron homeostasis via upregulating iron influx transporter TfR1 and downregulating iron efflux transporter FPN, thus leading to chondrocytes iron overload. Iron overload would promote the expression of chondrocytes catabolic markers, MMP3 and MMP13 expression. In addition, we found that oxidative stress and mitochondrial dysfunction played important roles in iron overload-induced cartilage degeneration, reducing iron concentration using iron chelator or antioxidant drugs could inhibit iron overload-induced OA-related catabolic markers and mitochondrial dysfunction. Our results suggest that pro-inflammatory cytokines could disrupt chondrocytes iron homeostasis and promote iron influx, iron overload-induced oxidative stress and mitochondrial dysfunction play important roles in iron overload-induced cartilage degeneration.


Assuntos
Condrócitos/patologia , Citocinas/toxicidade , Mediadores da Inflamação/toxicidade , Inflamação/complicações , Sobrecarga de Ferro/complicações , Osteoartrite/patologia , Estresse Oxidativo , Animais , Células Cultivadas , Condrócitos/metabolismo , Masculino , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/genética , Metaloproteinase 3 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Osteoartrite/etiologia , Osteoartrite/metabolismo
4.
Chin Med J (Engl) ; 134(8): 963-970, 2021 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-33840739

RESUMO

BACKGROUND: Histone deacetylase 4 (HDAC4) regulates chondrocyte hypertrophy and bone formation. The aim of the present study was to explore the effects of HDAC4 on Interleukin 1 beta (IL-1ß)-induced chondrocyte extracellular matrix degradation and whether it is regulated through the WNT family member 3A (WNT3A)/ß-catenin signaling pathway. METHODS: Primary chondrocytes (CC) and human chondrosarcoma cells (SW1353 cells) were treated with IL-1ß and the level of HDAC4 was assayed using Western blotting. Then, HDAC4 expression in the SW1353 cells was silenced using small interfering RNA to detect the effect of HDAC4 knockdown on the levels of matrix metalloproteinase 3 (MMP3) and MMP13 induced by IL-1ß. After transfection with HDAC4 plasmids, the overexpression efficiency was examined using Real-time quantitative polymerase chain reaction (qRT-PCR) and the levels of MMP3 and MMP13 were assayed using Western blotting. After incubation with IL-1ß, the translocation of ß-catenin into the nucleus was observed using immunofluorescence staining in SW1353 cells to investigate the activation of the WNT3A/ß-catenin signaling pathway. Finally, treatment with WNT3A and transfection with glycogen synthase kinase 3 beta (GSK3ß) plasmids were assessed for their effects on HDAC4 levels using Western blotting. RESULTS: IL-1ß downregulated HDAC4 levels in chondrocytes and SW1353 cells. Furthermore, HDAC4 knockdown increased the levels of MMP3 and MMP13, which contributed to the degradation of the extracellular matrix. Overexpression of HDAC4 inhibited IL-1ß-induced increases in MMP3 and MMP13. IL-1ß upregulated the levels of WNT3A, and WNT3A reduced HDAC4 levels in SW1353 cells. GSK-3ß rescued IL-1ß-induced downregulation of HDAC4 in SW1353 cells. CONCLUSION: HDAC4 exerted an inhibitory effect on IL-1ß-induced extracellular matrix degradation and was regulated partially by the WNT3A/ß-catenin signaling pathway.


Assuntos
Condrócitos , Histona Desacetilases , Via de Sinalização Wnt , beta Catenina , Linhagem Celular Tumoral , Células Cultivadas , Condrócitos/metabolismo , Glicogênio Sintase Quinase 3 beta/genética , Histona Desacetilases/genética , Humanos , Interleucina-1beta/farmacologia , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz , Proteínas Repressoras , Proteína Wnt3A/genética , beta Catenina/genética , beta Catenina/metabolismo
5.
Aging (Albany NY) ; 13(7): 9646-9664, 2021 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-33744859

RESUMO

In this study, we using the in vivo destabilization of the medial meniscus (DMM) mouse model to investigate the role of bone morphogenetic protein 5 (BMP5) in osteoarthritis (OA) progression mediated via chondrocyte senescence and apoptosis. BMP5 expression was significantly higher in knee articular cartilage tissues of OA patients and DMM model mice than the corresponding controls. The Osteoarthritis Research Society International scores based on histological staining of knee articular cartilage sections were lower in DMM mice where BMP5 was knocked down in chondrocytes than the corresponding controls 4 weeks after DMM surgery. DMM mice with BMP5-deficient chondrocytes showed reduced levels of matrix-degrading enzymes such as MMP13 and ADAMTS5 as well as reduced cartilage destruction. BMP5 knockdown also decreased chondrocyte apoptosis and senescence by suppressing the activation of p38 and ERK MAP kinases. These findings demonstrate that BMP5 silencing inhibits chondrocyte senescence and apoptosis as well as OA progression by downregulating activity in the p38/ERK signaling pathway.


Assuntos
Apoptose/fisiologia , Proteína Morfogenética Óssea 5/metabolismo , Senescência Celular/fisiologia , Condrócitos/metabolismo , Osteoartrite/metabolismo , Proteína ADAMTS5/genética , Proteína ADAMTS5/metabolismo , Animais , Proteína Morfogenética Óssea 5/genética , Cartilagem Articular/metabolismo , Linhagem Celular , Progressão da Doença , Inativação Gênica , Humanos , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Camundongos , Osteoartrite/genética
6.
Cancer Res ; 81(9): 2415-2428, 2021 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-33526510

RESUMO

Multiple myeloma promotes systemic skeletal bone disease that greatly contributes to patient morbidity. Resorption of type I collagen-rich bone matrix by activated osteoclasts results in the release of sequestered growth factors that can drive progression of the disease. Matrix metalloproteinase-13 (MMP13) is a collagenase expressed predominantly in the skeleton by mesenchymal stromal cells (MSC) and MSC-derived osteoblasts. Histochemical analysis of human multiple myeloma specimens also demonstrated that MMP13 largely localizes to the stromal compartment compared with CD138+ myeloma cells. In this study, we further identified that multiple myeloma induces MMP13 expression in bone stromal cells. Because of its ability to degrade type I collagen, we examined whether bone stromal-derived MMP13 contributed to myeloma progression. Multiple myeloma cells were inoculated into wild-type or MMP13-null mice. In independent in vivo studies, MMP13-null mice demonstrated significantly higher overall survival rates and lower levels of bone destruction compared with wild-type controls. Unexpectedly, no differences in type I collagen processing between the groups were observed. Ex vivo stromal coculture assays showed reduced formation and activity in MMP13-null osteoclasts. Analysis of soluble factors from wild-type and MMP13-null MSCs revealed decreased bioavailability of various osteoclastogenic factors including CXCL7. CXCL7 was identified as a novel MMP13 substrate and regulator of osteoclastogenesis. Underscoring the importance of host MMP13 catalytic activity in multiple myeloma progression, we demonstrate the in vivo efficacy of a novel and highly selective MMP13 inhibitor that provides a translational opportunity for the treatment of this incurable disease. SIGNIFICANCE: Genetic and pharmacologic approaches show that bone stromal-derived MMP13 catalytic activity is critical for osteoclastogenesis, bone destruction, and disease progression. GRAPHICAL ABSTRACT: http://cancerres.aacrjournals.org/content/canres/81/9/2415/F1.large.jpg.


Assuntos
Metaloproteinase 13 da Matriz/metabolismo , Mieloma Múltiplo/metabolismo , Mieloma Múltiplo/mortalidade , Osteólise/genética , Transdução de Sinais/genética , Animais , Diferenciação Celular/genética , Linhagem Celular Tumoral , Quimiocinas CXC/metabolismo , Modelos Animais de Doenças , Progressão da Doença , Feminino , Humanos , Masculino , Metaloproteinase 13 da Matriz/genética , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Mieloma Múltiplo/genética , Mieloma Múltiplo/patologia , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteogênese/genética , Taxa de Sobrevida
7.
Int J Mol Sci ; 22(4)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572320

RESUMO

Osteoarthritis (OA) is a common degenerative disease characterized by the destruction of articular cartilage and chronic inflammation of surrounding tissues. Matrix metalloproteinase-13 (MMP-13) is the primary MMP involved in cartilage degradation through its particular ability to cleave type II collagen. Hence, it is an attractive target for the treatment of OA. However, the detailed molecular mechanisms of OA initiation and progression remain elusive, and, currently, there are no interventions available to restore degraded cartilage. This review fully illustrates the involvement of MMP-13 in the initiation and progression of OA through the regulation of MMP-13 activity at the molecular and epigenetic levels, as well as the strategies that have been employed against MMP-13. The aim of this review is to identify MMP-13 as an attractive target for inhibitor development in the treatment of OA.


Assuntos
Cartilagem Articular/patologia , Metaloproteinase 13 da Matriz/metabolismo , Inibidores de Metaloproteinases de Matriz/farmacologia , Osteoartrite/tratamento farmacológico , Cartilagem Articular/efeitos dos fármacos , Domínio Catalítico , Colágeno Tipo II/metabolismo , Cristalografia por Raios X , Progressão da Doença , Desenvolvimento de Medicamentos , Epigênese Genética/efeitos dos fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/ultraestrutura , Inibidores de Metaloproteinases de Matriz/uso terapêutico , Terapia de Alvo Molecular/métodos , Osteoartrite/genética , Osteoartrite/patologia
8.
J Bone Miner Metab ; 39(4): 534-546, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33569722

RESUMO

INTRODUCTION: To investigate the role of LncRNA PVT1 (plasmacytoma variant translocation 1) in hyperglycemia-triggered cartilage damage using the diabetic osteoarthritis (OA) mice model. MATERIALS AND METHODS: Streptozotocin (STZ) was used to induce mouse diabetes. Knee OA model was induced through transection of anterior cruciate ligament (ACLT). Severity of arthritis was assessed histologically by Safranin O-Fast Green Staining using Mankin Scores. LncRNA PVT1 and miR-146a were detected by real-time polymerase chain reaction (PCR) in cartilage tissue. Moreover, the interaction among PVT1, miR-146a, and SMAD4 was examined by luciferase reporter assays. Mice were injected intra-articularly with ad-siRNA-PVT1 and ad-siRNA scramble control. Articular concentrations of TNF-α, IL-1, IL-6 and TGF-ß1 were determined using enzyme-linked immunosorbent assay. Levels of type II Collagen (COL2A1), TGF-ß1, p-SMAD2, SMAD2, p-SMAD3, SMAD3, SMAD4 and nuclear SMAD4 were detected by western blot analysis. RESULTS: PVT1 expression was significantly increased, whereas miR-146a was markedly decreased in diabetic OA mice than in non-diabetic OA and control. Increased PVT1 expression in diabetic OA mice was significantly associated with Mankin score and reduced miR-146a as well as Collagen alpha-1(II) (COL2A1) expressions. In vivo, intra-articular injection of ad-siRNA-PVT1 efficiently increased miR-146a and COL2A1 expressions, alleviated joint inflammation, decreased the expression of pro-inflammatory mediators, and suppressed TGF-ß/SMAD4 pathway in diabetic OA mice. CONCLUSIONS: Our results demonstrate LncRNA PVT1 is involved in cartilage degradation in diabetic OA and correlated with disease severity. Efficiency of ad-siRNA-PVT1 in controlling joint inflammation in diabetic OA mice is associated with the suppression of the expression of miR-146a, pro-inflammatory cytokines and activation of TGF-ß/SMAD4 pathway.


Assuntos
Cartilagem Articular/patologia , Diabetes Mellitus Experimental/genética , Regulação para Baixo , MicroRNAs/genética , Osteoartrite/genética , RNA Longo não Codificante/metabolismo , Proteína Smad4/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Adenoviridae/metabolismo , Animais , Sequência de Bases , Cartilagem Articular/metabolismo , Colágeno Tipo II/metabolismo , Regulação para Baixo/genética , Células HEK293 , Humanos , Hiperglicemia/complicações , Hiperglicemia/patologia , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 3 da Matriz/metabolismo , Camundongos Endogâmicos C57BL , MicroRNAs/metabolismo , Osteoartrite/patologia , RNA Longo não Codificante/genética , RNA Interferente Pequeno/metabolismo , Índice de Gravidade de Doença , Transdução de Sinais , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
9.
Mol Cell Biochem ; 476(6): 2465-2478, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33604811

RESUMO

Matrix metalloproteinases (MMPs) play key roles in epithelial-mesenchymal transition (EMT) for the development of cancer cell invasion and metastasis. MMP-13 is an extracellular matrix (ECM)-degrading enzyme that plays crucial roles in angiogenesis, cell cycle regulation, niche maintenance, and transforming squamous epithelial cells in various tissues. CD44, a transmembrane glycoprotein expressed on esophageal tumor cells, is required for EMT induction and invasion in esophageal squamous cell carcinoma (ESCC). The transcription factor TWIST1, as EMT and stemness marker, regulates MMPs expression and is identified as the downstream target of CD44. In this study, we aimed to investigate the probable interplay between the expression of key genes contributing to ESCC development, including MMP-13, TWIST1, and CD44 with clinical features for introducing novel diagnostic and therapeutic targets in the disease. The gene expression profiling of MMP-13, TWIST1, and CD44 was performed using quantitative real-time PCR in tumor tissues from 50 ESCC patients compared to corresponding margin non-tumoral tissues. Significant overexpression of MMP-13, CD44S, CD44V3, CD44V6, and TWIST1 were observed in 74%, 36%, 44%, 44%, and 52% of ESCC tumor samples, respectively. Overexpression of MMP-13 was associated with stage of tumor progression, metastasis, and tumor location (P < 0.05). There was a significant correlation between TWIST1 overexpression and grade (P < 0.05). Furthermore, overexpression of CD44 variants was associated with stage of tumor progression, grade, tumor invasion, and location (P < 0.05). The results indicated the significant correlation between concomitant expression of MMP-13/TWIST1, TWIST1/CD44, and CD44/MMP-13 with each other in a variety of clinicopathological traits, including depth of tumor invasion, tumor location, stage of tumor, and lymph node involvement in ESCC tissue samples (P < 0.05). Collectively, our results indicate that the TWIST1-CD44-MMP-13 axis is involved in tumor aggressiveness, proposing these genes as regulators of EMT, diagnostic markers, and therapeutic targets in ESCC.


Assuntos
Transição Epitelial-Mesenquimal , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago/metabolismo , Receptores de Hialuronatos/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Proteínas de Neoplasias/metabolismo , Proteínas Nucleares/metabolismo , Proteína 1 Relacionada a Twist/metabolismo , Idoso , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patologia , Carcinoma de Células Escamosas do Esôfago/genética , Carcinoma de Células Escamosas do Esôfago/patologia , Feminino , Humanos , Receptores de Hialuronatos/genética , Masculino , Metaloproteinase 13 da Matriz/genética , Pessoa de Meia-Idade , Proteínas de Neoplasias/genética , Proteínas Nucleares/genética , Proteína 1 Relacionada a Twist/genética
10.
PLoS One ; 16(2): e0240642, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33626093

RESUMO

The presented experiment focuses on assessing the impact of HMB (hydroxy-ß-methobutyrate) supplementation of mothers during pregnancy on the development of the skeletal system of their offspring. For this purpose, an experiment was carried out on 12 clinically healthy sows of the Great White Poland breed, which were divided randomly into two groups the control and the HMB group. All animals were kept under standard conditions and received the same feed for pregnant females. In contrast, females from the HMB group between 70 and 90 days were supplemented with 3-hydroxy-3-methylbutyle in the amount of 0.2g/kg b.w/day. Immediately after birth, the piglets were also divided into groups based on: sex, and presence or lack HMB supplementation, and subsequently were euthanized and humerus bones from all piglets were collected. Mother's HMB supplementation during pregnancy affected the multiple index of their offspring. The higher humerus mass and length was observed with the greater effect in males. Maternal supplementation also influenced on the geometrical and mechanical properties of the humerus as in the case of mass, this effect was higher in males. Also, the collagen structure of the compacted and trabecular bone changed under the HMB addition. Maternal supplementation also affected the expression of selected proteins in growth cartilage and trabecular bone. The obtained results show that the administration to the mother during pregnancy by the HMB significantly affects the development of the humerus in many ways. The obtained results also confirm the utility of such experiments in understanding of the importance of the pregnancy diet as an develop and adaptable factor of offspring organisms and are the base for further research in that area as well as in the protein markers expression area.


Assuntos
Úmero/efeitos dos fármacos , Suínos/embriologia , Valeratos/farmacologia , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Animais Recém-Nascidos/embriologia , Animais Recém-Nascidos/metabolismo , Proteína Morfogenética Óssea 2/metabolismo , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/embriologia , Cartilagem , Dieta/veterinária , Suplementos Nutricionais , Feminino , Úmero/embriologia , Masculino , Exposição Materna , Metaloproteinase 13 da Matriz/metabolismo , Polônia , Gravidez , Inibidor Tecidual de Metaloproteinase-2/metabolismo , Valeratos/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
11.
Biochem Biophys Res Commun ; 544: 38-43, 2021 03 12.
Artigo em Inglês | MEDLINE | ID: mdl-33516880

RESUMO

Cobalt ions are the main wear particles associated with orthopaedic implants, causing adverse complications due to cytotoxicity and inflammatory mediators. Recent studies have shown that sub-toxic levels of cobalt ions regulate matrix synthesis and inflammation, but the influence of cobalt ions on mechanotransduction remains unclear. Previously, we reported that sub-toxic levels of cobalt ions modulated primary cilia, which are crucial for mechanotransduction. This study therefore aimed to investigate the effect of cobalt ions on chondrocyte mechanosensation in response to cyclic tensile strain and the association with primary cilia. Sub-toxic levels of cobalt ions impaired chondrocyte mechanosensation and affected the gene expression of aggrecan, collagen II and MMP-13. Moreover, cobalt ions induced HDAC6-dependent primary cilia disassembly, which was associated with either cytoplasmic or ciliary α-tubulin deacetylation. Pharmaceutical HDAC6 inhibition with tubacin restored primary cilia length and mechanotransduction, whereas chemical depletion of primary cilia by chloral hydrate prevented mechanosignalling. Thus, sub-toxic levels of cobalt ions impaired chondrocyte mechanotransduction via HDAC6 activation, which was associated with tubulin deacetylation and primary cilia shortening.


Assuntos
Anilidas/farmacologia , Condrócitos/patologia , Cílios/patologia , Cobalto/toxicidade , Ácidos Hidroxâmicos/farmacologia , Metaloproteinase 13 da Matriz/metabolismo , Mecanotransdução Celular , Animais , Bovinos , Células Cultivadas , Condrócitos/efeitos dos fármacos , Condrócitos/metabolismo , Cílios/efeitos dos fármacos , Cílios/metabolismo , Inibidores de Histona Desacetilases/farmacologia , Oligoelementos/toxicidade
12.
Biomolecules ; 11(2)2021 01 20.
Artigo em Inglês | MEDLINE | ID: mdl-33498283

RESUMO

Malignant melanoma favors spreading to bone, resulting in a weakened bone with a high fracture risk. Here, we revealed the disorganized alignment of apatite crystals in the bone matrix associated with the homing of cancer cells by developing an artificially controlled ex vivo melanoma bone metastasis model. The ex vivo metastasis model reflects the progressive melanoma cell activation in vivo, resulting in decreased bone mineral density and expression of MMP1-positive cells. Moreover, less organized intercellular connections were observed in the neighboring osteoblasts in metastasized bone, indicating the abnormal and randomized organization of bone matrix secreted by disconnected osteoblasts. Our study revealed that the deteriorated microstructure associated with disorganized osteoblast arrangement was a determinant of malignant melanoma-related bone dysfunction.


Assuntos
Neoplasias Ósseas/metabolismo , Osso e Ossos/metabolismo , Melanoma/metabolismo , Metástase Neoplásica , Osteoblastos/metabolismo , Neoplasias Cutâneas/metabolismo , Animais , Anisotropia , Neoplasias Ósseas/secundário , Progressão da Doença , Fêmur/metabolismo , Metaloproteinase 13 da Matriz/metabolismo , Melanoma Experimental , Camundongos , Risco , Difração de Raios X , Microtomografia por Raio-X
13.
J Orthop Surg Res ; 16(1): 55, 2021 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-33446250

RESUMO

BACKGROUND: Intervertebral disk degeneration (IDD) is a degenerative disease characterized by cytoplasm loss and extracellular matrix degradation. Numerous evidence reported that miRNAs participated in IDD development. Nevertheless, the function of miR-142-3p in IDD development remains unknown. This study mainly explored the potential role and function of miR-142-3p in IDD development. METHODS: One percent fetal bovine serum was used to induce the degeneration of ATDC5 cells, and miR-142-3p level was examined by qRT-PCR. Then, miR-142-3p mimic/inhibitor and its corresponding negative control were transfected into ATDC5 normal and degenerative cells. Viability, migration, invasion, apoptosis, cycle, Bax, Bcl-2, P62, and Beclin1 expression levels were assessed using CCK8, wound healing assay, annexin V-FITC/PI staining, western blot, and qRT-PCR, respectively. RESULTS: The results revealed that the expression levels of MMP13, ADAMTS5, MMP3, and Col-X were increased as well as the expression levels of SOX-9 and Col-II were reduced in ATDC5 degenerative cells, indicating the degeneration model was constructed. We observed that miR-142-3p was decreased in ATDC5 degenerative cells and its suppression could promote ATDC5 cell degeneration. However, miR-142-3p overexpression could reverse the cell viability inhibition, as well as apoptosis and autophagy enhancement in ATDC5 degenerative cells. CONCLUSIONS: Our results proved that miR-142-3p may play an important role in disk degeneration. Further animal study is needed to illustrate the role of the miR-142-3p in IDD development.


Assuntos
Expressão Gênica , Estudos de Associação Genética , Degeneração do Disco Intervertebral/genética , MicroRNAs/genética , MicroRNAs/fisiologia , Proteína ADAMTS5/genética , Proteína ADAMTS5/metabolismo , Apoptose/genética , Autofagia/genética , Movimento Celular/genética , Sobrevivência Celular/genética , Células Cultivadas , Regulação para Baixo/genética , Humanos , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , MicroRNAs/metabolismo
14.
Int J Mol Sci ; 22(3)2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33513878

RESUMO

The invasion of extravillous trophoblast (EVT) cells into the maternal decidua, which plays a crucial role in the establishment of a successful pregnancy, is highly orchestrated by a complex array of regulatory mechanisms. Non-coding RNAs (ncRNAs) that fine-tune gene expression at epigenetic, transcriptional, and post-transcriptional levels are involved in the regulatory mechanisms of EVT cell invasion. However, little is known about the characteristic features of EVT-associated ncRNAs. To elucidate the gene expression profiles of both coding and non-coding transcripts (i.e., mRNAs, long non-coding RNAs (lncRNAs), and microRNAs (miRNAs)) expressed in EVT cells, we performed RNA sequencing analysis of EVT cells isolated from first-trimester placentae. RNA sequencing analysis demonstrated that the lncRNA H19 and its derived miRNA miR-675-5p were enriched in EVT cells. Although miR-675-5p acts as a placental/trophoblast growth suppressor, there is little information on the involvement of miR-675-5p in trophoblast cell invasion. Next, we evaluated a possible role of miR-675-5p in EVT cell invasion using the EVT cell lines HTR-8/SVneo and HChEpC1b; overexpression of miR-675-5p significantly promoted the invasion of both EVT cell lines. The transcription factor gene GATA2 was shown to be a target of miR-675-5p; moreover, small interfering RNA-mediated GATA2 knockdown significantly promoted cell invasion. Furthermore, we identified MMP13 and MMP14 as downstream effectors of miR-675-5p/GATA2-dependent EVT cell invasion. These findings suggest that miR-675-5p-mediated GATA2 inhibition accelerates EVT cell invasion by upregulating matrix metalloproteinases.


Assuntos
Fator de Transcrição GATA2/antagonistas & inibidores , Metaloproteinases da Matriz/metabolismo , MicroRNAs/metabolismo , Placenta/metabolismo , RNA Longo não Codificante/metabolismo , Trofoblastos/metabolismo , Linhagem Celular , Movimento Celular/genética , Movimento Celular/fisiologia , Proliferação de Células/genética , Proliferação de Células/fisiologia , Feminino , Fator de Transcrição GATA2/genética , Fator de Transcrição GATA2/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Metaloproteinase 14 da Matriz/genética , Metaloproteinase 14 da Matriz/metabolismo , Metaloproteinases da Matriz/genética , MicroRNAs/genética , Gravidez , Primeiro Trimestre da Gravidez , RNA Longo não Codificante/genética , RNA Interferente Pequeno , RNA-Seq , Trofoblastos/enzimologia
15.
Lasers Med Sci ; 36(3): 475-484, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32833088

RESUMO

Osteoarthritis (OA) is a chronic degenerative joint disease and is considered as the most common cause of pain and disability. To the best of our knowledge, it is generally observed that there is a lack of evidence on the effects of low-level laser therapy (LLLT) on inflammatory cytokines in OA. The present review aims to appraise the current evidence of efffects of LLLT on inflammatory cytokines in OA of the knee. Medical databases such as Medline, PubMed, EMBASE, PEDro CINAHL, Web of Science, Cochrane register, and Google reference were searched from its inception to June 2019. Articles that meet the inclusion criteria: subjects (animals-Wistar rats) induced with OA; rats with age group of 50-90 days; weight of 150-300 g; finding the effects of LLLT; reporting inflammatory cytokines; and articles written in English were included. The reviewers assessed the methodological quality of the primary studies. Data of inflammatory cytokines IL-1ß, IL-6, TNF-α, and MMP-13 were extracted for analysis. The Q (x2) test and I2 statistics analysis were performed to find the heterogeneity evaluation. Standard mean difference (SMD) and its 95% confidence interval (CI) were used to synthesize the data. Two hundred eleven potential articles were identified and 186 articles were excluded based on the selection criteria. The rest of the 25 articles were read and 8 articles were selected for further study. From the study, it is observed that the laser therapy group had mild to moderate improvement than control group in IL-1ß, TNF-α, and MMP-13 (IL-1ß; SMD 1.21 [95% CI - 0.278, 2.704], TNF-α; SMD 5.19 [95% CI 2.413, 7.961], and MMP-13 SMD - 1.45 [95% CI - 5.121, 2.211]), while IL-6 [SMD 3.11 (95% CI 0.662, 5.549] did not show any considerable improvement after laser therapy. The present review provides the evidence of LLLT-dependent reduction of IL-1ß, TNF-α, and MMP-13, and its ability to modulate proliferation of inflammatory cells, which makes LLLT a suitable treatment for OA. Though the included studies showed a high heterogeneity in treatment parameter, the beneficial effect of LLLT on changes in inflammatory cytokines, such as IL-6, seems to be unaffected.


Assuntos
Biomarcadores/metabolismo , Mediadores da Inflamação/metabolismo , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Terapia com Luz de Baixa Intensidade , Metaloproteinase 13 da Matriz/metabolismo , Osteoartrite/radioterapia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Modelos Animais de Doenças , Masculino , Viés de Publicação , Ratos Wistar
16.
J Orthop Res ; 39(3): 543-552, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-32716572

RESUMO

This study aimed to examine the effects of an episode of in vivo cyclic loading on rat knee articular cartilage (AC) under medium-term observation, while also investigating relevant factors associated with the progression of post-traumatic osteoarthritis (PTOA). Twelve-week-old Wistar rats underwent one episode comprising 60 cycles of 20 N or 50 N dynamic compression on the right knee joint. Spatiotemporal changes in the AC after loading were evaluated using histology and immunohistochemistry at 3 days and 1, 2, 4, and 8 weeks after loading (n = 6 for each condition). Chondrocyte vitality was assessed at 1, 3, 6, and 12 hours after loading (n = 2 for each condition). A localized AC lesion on the lateral femoral condyle was confirmed in all subjects. The surface and intermediate cartilage in the affected area degenerated after loading, but the calcified cartilage remained intact. Expression of type II collagen in the lesion cartilage was upregulated after loading, whereas the superficial lubricin layer was eroded in response to cyclic compression. However, the distribution of superficial lubricin gradually recovered to the normal level 4 weeks after loading-induced injury. We confirmed that 60 repetitions of cyclic loading exceeding 20 N could result in cartilage damage in the rat knee. Endogenous repairs in well-structured joints work well to rebuild protective layers on the lesion cartilage surface, which may be the latent factor delaying the progression of PTOA.


Assuntos
Cartilagem Articular/fisiologia , Condrócitos/fisiologia , Colágeno Tipo II/metabolismo , Articulação do Joelho/fisiologia , Proteoglicanas/metabolismo , Proteína ADAMTS5/metabolismo , Animais , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Distribuição Aleatória , Ratos Wistar , Suporte de Carga
17.
Lasers Med Sci ; 36(3): 529-540, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32519204

RESUMO

The presence of intra-articular crystals is detected in different articular pathologies of acute or chronic nature. The aim of this work was to analyze the action of the indium gallium aluminum and phosphorus (InGaAlP) (λ = 670 nm) laser on the synovial membrane present in the knee joint in experimentally induced microcrystalline arthritis in male adult Wistar rats. The animals were divided into three experimental groups (n = 24): control (A), experimentally induced arthritis (B), experimentally induced arthritis+InGaAlP laser therapy (C). The laser treatment was made daily in the patellar region of the right knee after 48 h of the experimental induction. After 7, 14, and 21 days of therapy, the rats were euthanized and the right knees were removed and processed for histomorphometric, immunohistochemical, ultrastructural, and biochemical investigation of the synovium. The number of granulocytes on the 14th and 21st days was higher in B and lower in C and, lastly, in A. The number of fibroblasts on the 14th and 21st days was similar between A and C and below B. The number of blood vessels on the 21st day was higher in B than in the other groups. The positive number of cells for the TUNEL test was higher on the 14th and 21st days in B compared to the others. The percentage of tissue area occupied by birefringent collagen fibers was higher in B on the 21st day than in the others. The ultrastructure of cells showed fibroblast-like morphology in all groups and periods evaluated. The quantification of glycosaminoglycans did not present significant differences between the groups in all the experimental periods. The amount of hydroxyproline was higher in B compared to the other groups on the 14th and 21st days. The content of non-collagen proteins was higher in B on the 21st day in relation to the other groups. Quantification of TNF-α on the 21st day was higher in A and B than in C. For TGF-ß on the 21st day, groups B and C presented similar and higher values than A. For MMP-13, groups A and B presented data similar to and above C. In relation to ADAMT-S4, on the 21st day, groups B and C presented data similar to and lower than A. InGaAlP-670 nm therapy reduced the inflammatory process and tissue injuries of the synovial membrane in comparison to the untreated group, indicating its potential utilization in clinical studies aiming in the recovery of acute arthritis in patients.


Assuntos
Artrite Experimental/cirurgia , Terapia a Laser , Membrana Sinovial/patologia , Membrana Sinovial/efeitos da radiação , Proteína ADAMTS4/metabolismo , Animais , Apoptose/efeitos da radiação , Vasos Sanguíneos/patologia , Vasos Sanguíneos/efeitos da radiação , Cristalização , Articulação do Joelho/patologia , Masculino , Metaloproteinase 13 da Matriz/metabolismo , Ratos Wistar , Membrana Sinovial/ultraestrutura , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
18.
J Ethnopharmacol ; 267: 113506, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33148433

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Alstonia scholaris (L.) R. Br. (Apocynaceae) is a Dai folk medicine for the treatment of lung diseases in China. AIM OF THE STUDY: The present study investigated the anti-pulmonary fibrosis effects of total alkaloids (TA) and the potential active ingredients and its possible mechanism. MATERIALS AND METHODS: After intratracheal instillation of bleomycin (BLM, 5 mg/kg), mice were divided into ten groups, and orally treated with the corresponding samples once daily for 28 days. The effect of indole alkaloids was determined through analysis of cytokines, as well as histopathological examinations and gene expressions. RESULTS: Severe lung fibrosis was observed in the BLM-treated mice on day 28. However, the administration of TA significantly ameliorated the pathological changes in the lungs, decreased the content of Krebs von den Lungen-6, lactate dehydrogenase, transforming growth factor-ß (TGF-ß), hydroxyproline, type I collagen, and malonaldehyde, and enhanced the activity of superoxide dismutase in the serum and lung tissues. In addition, the enhanced TGF-ß and matrix metalloproteinase-1 (MMP-1) expressions in BLM-induced mice were obviously weakened by indole alkaloids, as well as the ratio of matrix metalloproteinase-1 to tissue inhibitor of metalloproteinase-1 was decreased. Moreover, picrinine and scholaricine yielded markedly better values in the aforementioned indices than those in other samples, indicating that they may be the active ingredients of alkaloids. CONCLUSIONS: TA exerted protective effects against BLM-induced pulmonary fibrosis by reducing collagen deposition through TGF-ß/MMP-1 pathway.


Assuntos
Alstonia , Alcaloides Indólicos/farmacologia , Pulmão/efeitos dos fármacos , Extratos Vegetais/farmacologia , Fibrose Pulmonar/prevenção & controle , Alstonia/química , Animais , Bleomicina , Colágeno/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica , Alcaloides Indólicos/isolamento & purificação , Mediadores da Inflamação/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Masculino , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 13 da Matriz/metabolismo , Camundongos Endogâmicos ICR , Extratos Vegetais/isolamento & purificação , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Transdução de Sinais , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo
19.
Int Immunopharmacol ; 91: 107191, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33359852

RESUMO

This study aims to investigate the effects and mechanisms of parathyroid hormone [1-34] (PTH1-34) on TNF-α-stimulated mice chondrocytes, as well as cartilage from a meniscus injury induced osteoarthritis (MIO) mice model. The C57BL/6J mice received medial meniscectomy, and then administrated with PTH1-34. The results showed that PTH1-34 administration decreased secondary allodynia and the pain-related transcripts. The IHC, ELISA, Micro-CT imaging and histopathology analysis revealed the significantly improved subchondral plate thickness and bone porosity, the reduced pro-inflammatory cytokines in serum and joint fluid. In vitro, mice chondrocyte was treated with TNF-α or co-cultured with synovial cells. The results showed that TNF-α markedly upregulated the MMP13 expression, and the ERK1/2, NF-κB or PI3K signaling pathway inhibitors could reverse the induction effect of TNF-α on expression of MMP13 in chondrocytes. PTH1-34 alone has no effect on the expression of MMP13 and NF-κB signaling pathways, but the PTH1-34 could reverse the induction effect of TNF-α on MMP13 expression and NF-κB signaling pathway activation in chondrocytes. In addition, PTH1-34 administration inhibited the expression of TNF-α and MMP13, and chondrocyte viability, while the PKA repressor reversed the effect of PTH1-34 in chondrocytes co-cultured with synovial cells. In conclusion, PTH1-34 has an obvious analgesic and anti-inflammatory effect, inhibits the matrix synthesis and alleviates the progression of osteoarthritis. In vitro, PTH1-34 inhibited TNF-α expression and antagonized TNF-α-induced MMP13 expression via the PKA pathway and the NF-κB signaling pathways, respectively.


Assuntos
Analgésicos/farmacologia , Anti-Inflamatórios/farmacologia , Artralgia/prevenção & controle , Condrócitos/efeitos dos fármacos , Articulações/efeitos dos fármacos , Metaloproteinase 13 da Matriz/metabolismo , Menisco/efeitos dos fármacos , Osteoartrite/prevenção & controle , Teriparatida/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Animais , Artralgia/enzimologia , Artralgia/etiologia , Células Cultivadas , Condrócitos/enzimologia , Condrócitos/patologia , Técnicas de Cocultura , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Modelos Animais de Doenças , Articulações/enzimologia , Articulações/patologia , Meniscectomia , Menisco/enzimologia , Menisco/patologia , Menisco/cirurgia , Camundongos Endogâmicos C57BL , NF-kappa B/metabolismo , Osteoartrite/enzimologia , Osteoartrite/etiologia , Osteoartrite/patologia , Transdução de Sinais , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/enzimologia , Membrana Sinovial/patologia
20.
Int J Mol Sci ; 21(23)2020 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-33287358

RESUMO

Gremlin-1 (GREM1), one of the bone morphogenetic protein (BMP) antagonists, can directly bind to BMPs. GREM1 is involved in organogenesis, tissue differentiation, and organ fibrosis. Recently, numerous studies have reported the oncogenic role of GREM1 in cancer. However, the role of GREM1 in metastasis of breast cancer cells and its underlying mechanisms remain poorly understood. The role of GREM1 in breast cancer progression was assessed by measuring growth, migration, and invasion of breast cancer cells. An orthotopic breast cancer mouse model was used to investigate the role of GREM1 in lung metastasis of breast cancer cells. GREM1 knockdown suppressed the proliferation of breast cancer cells, while its overexpression increased their growth, migration, and invasion. Cells with Grem1-knockdown showed much lower tumor growth rates and lung metastasis than control cells. GREM1 enhanced the expression of matrix metalloproteinase 13 (MMP13). A positive correlation between GREM1 and MMP13 expression was observed in breast cancer patients. GREM1 activated signal transducer and activator of transcription 3 (STAT3) transcription factor involved in the expression of MMP13. Our study suggests that GREM1 can promote lung metastasis of breast cancer cells through the STAT3-MMP13 pathway. In addition, GREM1 might be a promising therapeutic target for breast cancer metastasis.


Assuntos
Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/genética , Metaloproteinase 13 da Matriz/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais , Animais , Biomarcadores , Neoplasias da Mama/mortalidade , Neoplasias da Mama/patologia , Movimento Celular , Proliferação de Células , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Neoplasias Pulmonares/secundário , Metaloproteinase 13 da Matriz/genética , Camundongos , Prognóstico
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