Acupuncture Delays Cartilage Degeneration through Upregulating SIRT1 Expression in Rats with Osteoarthritis

Silent mating type information regulation 2 homolog 1 (SIRT1) has been reported to inhibit osteoarthritic gene expression in chondrocytes. Here, efforts in this study were made to unveil the specific role of SIRT1 in the therapy of acupuncture on cartilage degeneration in osteoarthritis (OA). Specifically, OA was established by the anterior cruciate ligament transection method in the right knee joint of rats, subsequent to which acupuncture was performed on two acupoints. Injection with shSIRT1 sequence–inserted lentiviruses was conducted to investigate the role of SIRT1 in acupuncture-mediated OA. Morphological changes and cell apoptosis in rat OA cartilages were examined by safranin-O staining and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay, respectively. The serum levels of tumor necrosis factor (TNF)-α and interleukin (IL)-2 in OA rats were assessed by enzyme-linked immunosorbent assay (ELISA). The expressions of SIRT1, cartilage matrix degradation-related proteins (matrix metalloproteinase (MMP)-9 and ADAMTS5), NF-κB signaling-related markers (p-p65/p65 and p-IκBα/IκBα), and cartilage matrix synthesis-related proteins (collagen II and aggrecan) in the OA cartilage were analyzed by western blot. As a result, acupuncture counteracted OA-associated upregulation of TNF-α, IL-2, cartilage matrix degradation-related proteins, and NF-κB signaling-related markers, morphological damage, apoptosis, SIRT1 downregulation, and loss of cartilage matrix synthesis-related proteins in rat articular cartilages. SIRT1 silencing reversed acupuncture-induced counteractive effects on the aforementioned OA-associated phenomena (except apoptosis, the experiment regarding which under SIRT1 silencing was not performed). Collectively, acupuncture inhibited chondrocyte apoptosis, inflammation, NF-κB signaling activation, and cartilage matrix degradation by upregulating SIRT1 expression to delay OA-associated cartilage degeneration.

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Fluoxetine suppresses inflammatory reaction in microglia under OGD/R challenge via modulation of NF-κB signaling

Bioscience Reports ◽

10.1042/bsr20181584 ◽

2019 ◽

Vol 39 (4) ◽

Cited By ~ 1

Author(s):

Mouli Tian ◽

Mei Yang ◽

Zhenjie Li ◽

Yiru Wang ◽

Wei Chen ◽

...

Keyword(s):

Ischemia Reperfusion Injury ◽

Ischemia Reperfusion ◽

Glucose Deprivation ◽

Cell Counting ◽

Enzyme Linked Immunosorbent Assay ◽

Oxygen Glucose Deprivation ◽

Tnf Α ◽

Anti Inflammatory ◽

Related Proteins

Abstract We aimed to investigate the anti-inflammatory role of fluoxetine, a selective serotonin reuptake inhibitor, in microglia (MG) and the mechanisms under oxygen glucose deprivation/reoxygenation (OGD/R). An OGD/R model on BV-2 cells was used for the study of microglia under ischemia/reperfusion injury in ischemic stroke. Lentiviral transfection was applied to knock down IκB-α. Enzyme-linked immunosorbent assay (ELISA) was used for detecting levels of TNF-α, IL-1β, and IL-6, and real-time PCR was used to assess the expression of IκB-α protein. Western blotting was applied to analyze NF-κB-signaling related proteins and Cell Counting Kit-8 (CCK-8) was used for assessing cell viability. Molecular docking and drug affinity responsive target stability (DARTS) assay were used for the detection of the interaction between IκB-α and fluoxetine. We found that fluoxetine decreased the levels of TNF-α, IL-1β, and IL-6 in supernatant as well as NF-κB subunits p65 and p50 in BV-2 cells under OGD/R. Fluoxetine significantly increased the level of IκB-α through the inhibition of IκB-α ubiquitylation and promoted the bonding of IκB-α and fluoxetine in BV-2 cells under OGD/R. Knocking down IκB-α attenuated the decreasing effect of TNF-α, IL-1β, and IL-6 as well as p65 and p50 in BV-2 cells under OGD/R led to by fluoxetine. In conclusion, our present study demonstrated the anti-inflammatory role of fluoxetine and its mechanisms related to the modulation of NF-κB-related signaling in MG under ischemia/reperfusion challenge.

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In vivo protective effects of upper zone of growth plate and cartilage matrix associated protein against cartilage degeneration in a monosodium iodoacetate induced osteoarthritis model

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2020-0009 ◽

2020 ◽

Vol 98 (11) ◽

pp. 763-770

Author(s):

Hamza Malik Okuyan ◽

Menderes Yusuf Terzi ◽

İhsan Karaboğa ◽

Serdar Doğan ◽

Aydıner Kalacı

Keyword(s):

Growth Plate ◽

Matrix Protein ◽

Critical Role ◽

Cartilage Degeneration ◽

Cartilage Matrix ◽

Bone Morphogenetic Protein 2 ◽

Monosodium Iodoacetate ◽

And Cartilage

Osteoarthritis (OA) is a degenerative disease affecting the majority of over 65 year old people and characterized by cartilage degeneration, subchondral abnormal changes, and inflammation. Despite the enormous socioeconomic burden caused by OA, currently, there is no effective therapy against it. Upper zone of growth plate and cartilage matrix associated protein (UCMA) is a vitamin K dependent protein and has a critical role in pathophysiological conditions associated with bone and cartilage. However, there is no research on the protective role of intra-articular UCMA treatment in OA pathogenesis. Therefore, we aimed to investigate the potential therapeutic role of UCMA in an in vivo model of OA. We report for the first time that intra-articular UCMA injection ameliorated cartilage degeneration in a monosodium iodoacetate induced OA rat model. Furthermore, the OA-induced activation of nuclear factor kappa B and bone morphogenetic protein 2 signals was attenuated by UCMA. Our results indicated that UCMA decreased cartilage oligomeric matrix protein levels but did not affect interleukin 6, total antioxidant status, and total oxidant status levels in the serum. In conclusion, UCMA exhibited a therapeutic potential in the treatment of OA. This protective effect of UCMA is possibly achieved by reducing the aggrecanase activity and the production of inflammatory cytokines.

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Paeonol Ameliorates Inflammation and Cartilage of Chondrocytes in Osteoarthritis by Upregulating SIRT1

10.21203/rs.3.rs-104024/v1 ◽

2020 ◽

Author(s):

Peng Shang ◽

Ying Liu ◽

Junqing Jia

Keyword(s):

Extracellular Matrix ◽

Caspase 3 ◽

Type Ii Collagen ◽

Blue Staining ◽

Chondrocyte Apoptosis ◽

Matrix Degradation ◽

Extracellular Matrix Degradation ◽

Tnf Α ◽

And Cartilage

Abstract Objective: To explore the possible role of paeonol on chondrocyte inflammation and cartilage protection in osteoarthritis (OA).Methods: Primary chondrocytes were isolated from rat stifle joints, and were identified through toluidine blue staining and immunofluorescence staining of type II collagen. The chondrocytes were transfected with sh-SIRT1 or/and paeonol (0, 20, 50, 100, 200, 1000 mg/L) before OA modeling induced by IL-1β. ELISA determined the expressions of TNF-α, IL-17 and IL-6, and apoptotic rate was examined by flow cytometry. qRT-PCR and Western blot quantified the expressions of MMP-1, MMP-3, MMP-13, TIMP-1, cleaved-caspase-3, Bax, Bcl-2, and the proteins related to NF-κB pathway.Results: Increases in TNF-α, IL-17, IL-6, MMP-1, MMP-3 and MMP-13 and decrease in TIMP-1 were found in IL-1β stimulated chondrocytes. The apoptotic rate as well as the expressions of cleaved-caspase-3 and Bax was up-regulated, and Bcl-2 expression was suppressed in response to IL-1β treatment. NF-κB pathway was activated in IL-1β-stimulated chondrocytes. Paeonol enhanced SIRT1 expression to inactivate NF-κB pathway, thus ameliorating the secretion of inflammatory cytokines, extracellular matrix degradation and chondrocyte apoptosis.Conclusion: Paeonol inhibits IL-1β induced inflammation and extracellular matrix degradation in chondrocytes through up-regulating SIRT1 and suppressing NF-κB pathway.

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Fibroblast-like Synoviocytes-derived Exosomal PCGEM1 Accelerates IL-1β-induced Apoptosis and Cartilage Matrix Degradation by miR-142-5p/RUNX2 in Chondrocytes

Immunological Investigations ◽

10.1080/08820139.2021.1936010 ◽

2021 ◽

pp. 1-18

Author(s):

Guangxuan Zeng ◽

Gang Deng ◽

Shiliang Xiao ◽

Fei Li

Keyword(s):

Cartilage Matrix ◽

Matrix Degradation ◽

Induced Apoptosis ◽

And Cartilage ◽

Fibroblast Like Synoviocytes

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Metabolic Syndrome Predisposes to Osteoarthritis: Lessons from Model System

Cartilage ◽

10.1177/1947603520980161 ◽

2020 ◽

pp. 194760352098016

Author(s):

Sampath Samuel Joshua Pragasam ◽

Vijayalakshmi Venkatesan

Keyword(s):

Subchondral Bone ◽

Fat Mass ◽

Articular Chondrocytes ◽

Bone Cyst ◽

Primary Cultures ◽

Abdominal Circumference ◽

Enzyme Linked Immunosorbent Assay ◽

Micro Ct ◽

Tnf Α ◽

And Cartilage

Objective The present study aims to assess for temporal changes in tibial subchondral bone and cartilage in WNIN/Gr-Ob rats (portraying obesity, insulin resistance, dyslipidemia, impaired glucose tolerance, hypertension) in comparison with Wistar controls (WNIN) using anthropometry, micro-computed tomography (micro-CT), scanning electron microscopy (SEM), histopathology, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence. Design Body weight, abdominal circumference, body mass index (BMI), lean/fat mass, serum tumor necrosis factor (TNF)-α levels were measured (ELISA), followed by ultrastructural analysis of tibial subchondral bone (micro-CT) and cartilage architecture (histopathology and SEM) in WNIN/Gr-Ob and WNIN rats with age (3, 6 and 9 months). Additionally, primary cultures of articular chondrocytes isolated from 6-month-old WNIN/Gr-Ob and WNIN rats were assessed for matrix metalloproteinase (MMP)-13 and Collagen type II (COL2A1) by immunofluorescence. Results WNIN/Gr-Ob rats exhibited frank obesity with increased BMI, lean and fat mass vis-à-vis significantly higher levels of serum TNF-α (6>9>3 months) as compared with the controls. With an increase in BMI, WNIN/Gr-Ob rats presented with tibial cartilage fibrillation, erosion, osteophyte formation (6 months) and subchondral bone cyst (9 months) confirmed by histology and SEM. An increase in subchondral trabecular bone volume (sclerosis with decreased plate porosity) was observed in all ages in WNIN/Gr-Ob rats compared to their Control. Gaining insights, primary cultures of articular chondrocytes complemented with altered cellular expressions of COL2A1 and MMP-13 from WNIN/Gr-Ob rats, indicating osteoarthritis (OA) progression. Conclusion Multiple metabolic perturbations featured in WNIN/Gr-Ob rats were effective to induce spontaneous OA-like degenerative changes affecting knee joints akin to human OA.

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