RETRACTED ARTICLE: Effects of Osteopontin on Expression of IL-6 and IL-8 Inflammatory Factors in Human Knee Osteoarthritis Chondrocytes

Abstract Background Circular RNA (circRNA) has been shown to be associated with osteoarthritis (OA) progression. Circ_0116061 has been found to be highly expressed in OA cartilage tissues, but its role and mechanism in OA progression remain unclear. Methods Expression levels of circ_0116061, microRNA (miR)-200b-5p, and Smad ubiquitin regulatory factor 2 (SMURF2) were detected using quantitative real-time PCR. The proliferation and apoptosis of cells were measured using cell counting kit 8 (CCK8) assay, colony formation assay, and flow cytometry. Furthermore, the protein levels of proliferation-related marker, apoptosis-related markers, inflammatory factors, and SMURF2 were tested using western blot (WB) analysis. In addition, the interaction between miR-200b-3p and circ_0116061 or SMURF2 was examined using dual-luciferase reporter assay and biotin-labeled RNA pull-down assay. Results Circ_0116061 and SMURF2 were highly expressed, and miR-200b-3p was lowly expressed in OA cartilage tissues. Knockdown of circ_0116061 could promote the proliferation and inhibit the apoptosis and inflammation of OA chondrocytes. MiR-200b-3p could be sponged by circ_0116061, and its inhibitor could reverse the regulation of circ_0116061 silencing on the biological functions of OA chondrocytes. SMURF2 was a target of miR-200b-3p, and its expression was positively regulated by circ_0116061. Silencing of SMURF2 also could enhance the proliferation and suppress the apoptosis and inflammation of OA chondrocytes. Furthermore, the regulation of circ_0116061 silencing on the biological functions of OA chondrocytes also could be reversed by SMURF2 overexpression. Conclusion Our data showed that circ_0116061 might regulate the miR-200b-3p/SMURF2 axis to promote the progression of OA.

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Immunofluorescence Analysis of NF-kB and iNOS Expression in Different Cell Populations during Early and Advanced Knee Osteoarthritis

International Journal of Molecular Sciences ◽

10.3390/ijms22126461 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6461

Author(s):

Marko Ostojic ◽

Ante Zevrnja ◽

Katarina Vukojevic ◽

Violeta Soljic

Keyword(s):

Blood Vessel ◽

Knee Osteoarthritis ◽

Dominant Role ◽

Inos Expression ◽

Inflammatory Factors ◽

Kb Cells ◽

Immunofluorescence Analysis ◽

Leading Role ◽

Radiological Score ◽

Early Oa

Synovitis of the knee synovium is proven to be a precursor of knee osteoarthritis (OA), leading to a radiologically advanced stage of the disease. This study was conducted to elucidate the expression pattern of different inflammatory factors—NF-kB, iNOS, and MMP-9 in a subpopulation of synovial cells. Thirty synovial membrane intra-operative biopsies of patients (ten controls, ten with early OA, and ten with advanced OA, according to the Kellgren–Lawrence radiological score) were immunohistochemically stained for NF-kB, iNOS, and MMP9, and for different cell markers for macrophages, fibroblasts, leukocytes, lymphocytes, blood vessel endothelial cells, and blood vessel smooth muscle cells. The total number of CD68+/NF-kB+ cells/mm2 in the intima of early OA patients (median = 2359) was significantly higher compared to the total number of vimentin+/Nf-kB+ cells/mm2 (median = 1321) and LCA+/NF-kB+ cells/mm2 (median = 64) (p < 0.001 and p < 0.0001, respectively). The total number of LCA+/NF-kB+ cells/mm2 in the subintima of advanced OA patients (median = 2123) was significantly higher compared to the total number of vimentin+/NF-kB+ cells/mm2 (median = 14) and CD68+/NF-kB+ cells/mm2 (median = 29) (p < 0.0001). The total number of CD68+/iNOS+ cells/mm2 in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/iNOS+ cells/mm2 and LCA+/iNOS+ cells/mm2 (p < 0.0001 and p < 0.001, respectively). The total number of CD68+/MMP-9+ cells/mm2 in the intima of both early and advanced OA patients was significantly higher compared to the total number of vimentin+/MMP-9+ cells/mm2 and CD5+/MMP-9+ cells/mm2 (p < 0.0001). Macrophages may have a leading role in OA progression through the NF-kB production of inflammatory factors (iNOS and MMP-9) in the intima, except in advanced OA, where leukocytes could have a dominant role through NF-kB production in subintima. The blocking of macrophageal and leukocyte NF-kB expression is a possible therapeutic target as a disease modifying drug.

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Effect of Moxibustion on the Intestinal Flora of Rats with Knee Osteoarthritis Induced by Monosodium Iodoacetate

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2020/3196427 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Yu Chen ◽

Jiuheng Lv ◽

Yejuan Jia ◽

Ruiqing Wang ◽

Zidi Zhang ◽

...

Keyword(s):

Treatment Group ◽

Knee Osteoarthritis ◽

High Throughput Sequencing ◽

Cartilage Damage ◽

Intestinal Flora ◽

Inflammatory Factors ◽

Knee Cartilage ◽

Model Group ◽

Monosodium Iodoacetate ◽

And Cartilage

In this study, a knee osteoarthritis (KOA) rat model induced by monosodium iodoacetate (MIA) was used to study the effect of moxibustion on improving knee cartilage damage and its effect on the intestinal flora. The experimental rats were divided into the normal group (N), model group (M), moxibustion treatment group (MS), and diclofenac sodium treatment group (DS). After 4 weeks, cartilage pathological damage in the knee joint was evaluated using hematoxylin-eosin and safranin O-fast green staining analysis. ELISAs and Western blots were used to detect the expression levels of IL-1β and TNF-α in the serum and cartilage, respectively. The total DNA of the fecal samples was extracted and subjected to high-throughput sequencing of the V3-V4 region of the 16S rRNA gene to analyze the changes in the intestinal flora. In the model group, the cartilage was obviously damaged, the expression levels of IL-1β and TNF-α in the serum and cartilage were increased, and the abundance and diversity of the intestinal flora were decreased. Moxibustion treatment significantly improved the cartilage damage and reduced the concentration of inflammatory factors in the serum and cartilage. The high-throughput sequencing results showed that compared to the model group, the moxibustion treatment regulated some specific species in the intestinal microorganisms rather than the α diversity. In conclusion, our findings suggest that moxibustion treatment may work through two aspects in rats. On one hand, it directly acts on knee cartilage to promote repair, and on the other hand, it regulates the composition of the intestinal flora and reduces the production of inflammatory factors.

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Effects of l-Carnitine Supplementation on Serum Inflammatory Factors and Matrix Metalloproteinase Enzymes in Females with Knee Osteoarthritis: A Randomized, Double-Blind, Placebo-Controlled Pilot Study

Journal of the American College of Nutrition ◽

10.1080/07315724.2015.1068139 ◽

2016 ◽

Vol 35 (7) ◽

pp. 597-603 ◽

Cited By ~ 17

Author(s):

Aida Malek Mahdavi ◽

Reza Mahdavi ◽

Sousan Kolahi

Keyword(s):

Pilot Study ◽

Matrix Metalloproteinase ◽

Knee Osteoarthritis ◽

Inflammatory Factors ◽

Carnitine Supplementation ◽

Double Blind ◽

Double Blind Placebo

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Platelet‑rich plasma shows beneficial effects for patients with knee osteoarthritis by suppressing inflammatory factors

Experimental and Therapeutic Medicine ◽

10.3892/etm.2018.5794 ◽

2018 ◽

Cited By ~ 8

Author(s):

Guilin Huang ◽

Sha Hua ◽

Tuanmin Yang ◽

Jianbing Ma ◽

Wenxing Yu ◽

...

Keyword(s):

Knee Osteoarthritis ◽

Platelet Rich Plasma ◽

Inflammatory Factors ◽

Beneficial Effects

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Repairing effects of glucosamine sulfate in combination with etoricoxib on articular cartilages of patients with knee osteoarthritis

10.21203/rs.3.rs-16773/v1 ◽

2020 ◽

Author(s):

Yong Sun ◽

Changde Wang ◽

Chunzhu Gong

Keyword(s):

Knee Osteoarthritis ◽

Womac Score ◽

Effective Rate ◽

Glucosamine Sulfate ◽

Inflammatory Factors ◽

Chondrocyte Apoptosis ◽

Mrna Levels ◽

Related Factors ◽

Articular Cartilages ◽

Experimental Group

Abstract Purpose: To evaluate the repairing effects of glucosamine sulfate combined with etoricoxib on articular cartilages of patients with knee osteoarthritis (KOA). Methods: A total of 106 KOA patients were randomly divided into control (n=40) and experimental groups (n=66), and treated with etoricoxib alone and glucosamine sulfate plus etoricoxib respectively. Changes in WOMAC score and clinical efficacy were observed. The synovial fluid was extracted. Bone metabolism indices, growth factors, inflammatory factors, matrix metalloproteinases (MMPs) and NO-induced apoptosis-related factors were measured by ELISA. JNK and Wnt5a mRNA levels were determined using RT-PCR. Results: After treatment, the total WOMAC scores of both groups significantly declined (P<0.05), being lower in experimental group. The total effective rate of experimental group was higher (P<0.05). BGP and OPG levels rose, especially in experimental group (P<0.05). CTX-II, COMP and RANKL levels decreased, particularly in experimental group (P<0.05). TGF-β, IGF-1 and FGF-2 levels increased, especially in experimental group (P<0.05). Both groups, particularly experimental group, had decreased levels of IL-1β, IL-17, IL-18, TNF-α, MMP-3, MMP-9 and MMP-13 (P<0.05). JNK and Wnt5a mRNA levels of both groups dropped, which were lower in experimental group (P<0.05). NO and LPO levels reduced, being lower in experimental group. SOD level rose, especially in experimental group (P<0.05). Conclusion: Glucosamine sulfate plus etoricoxib can repair the articular cartilages of KOA patients. Probably, JNK and Wnt5a are down-regulated to inhibit the secretion of MMPs through lowering the levels of inflammatory factors, thereby delaying cartilage matrix degradation. NO-induced chondrocyte apoptosis may be suppressed via the SOD pathway.

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Efficacy and Mechanism of Electroacupuncture Treatment of Rabbits With Different Degrees of Knee Osteoarthritis: A Study Based on Synovial Innate Immune Response

Frontiers in Physiology ◽

10.3389/fphys.2021.642178 ◽

2021 ◽

Vol 12 ◽

Author(s):

Anmin Ruan ◽

Qingfu Wang ◽

Yufeng Ma ◽

Dong Zhang ◽

Lili Yang ◽

...

Keyword(s):

Immune Response ◽

Western Blot ◽

Knee Osteoarthritis ◽

Synovial Tissue ◽

Innate Immune Response ◽

Synovial Membrane ◽

Innate Immune ◽

Cartilage Tissue ◽

Inflammatory Factors ◽

And Cartilage

Knee osteoarthritis (KOA) is a chronic degenerative bone and joint disease, which is often clinically manifested as pain, joint swelling, and deformity. Its pathological manifestations are mainly synovial inflammation and cartilage degeneration. This study aims to investigate the efficacy of electro-acupuncture (EA) on model rabbits with varying degrees of KOA and to study the mechanism of EA on KOA based on the innate immune response. Mild and moderate rabbit KOA models were established using a modified Hluth method, and EA was given to both the mild and moderate model groups. The Lequesne-MG index was used to evaluate the behavioral changes in the rabbits before and after EA treatment. Morphological changes in the synovial membrane and cartilage of each group were observed by H&E staining. The Mankin scoring standard and the Krenn scoring standard were used to score the pathology of the cartilage tissue and synovial tissue, respectively. The inflammatory factors and metalloproteinases were detected in the serum of each group by ELISA. The protein and messenger RNA (mRNA) expressions of important elements related to Toll-like receptors (TLRs)-mediated innate immune response in the synovial tissue were detected by Western blot and quantitative PCR (qPCR). The Lequesne-MG index score of the rabbits gradually increased with the modeling prolonged but decreased significantly after EA treatment, indicating that EA has a better effect on alleviating the pain and improving the dysfunction. The morphological analysis showed that the inflammation of and the damage to the synovial membrane and the cartilage tissue gradually deteriorated with the modeling prolonged. However, the synovial membrane inflammation was significantly relieved after EA treatment, and the cartilage injury showed signs of repair. The ELISA analysis showed that, with the modeling prolonged, the serum-related inflammatory factors and mechanism of metalloproteinases gradually increased but decreased after EA treatment. The tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), and matrix metalloproteinase3 (MMP3) of EA1 group were significantly lower than those of EA2 group. Both Western blot and qPCR results showed that the protein and mRNA expressions of the elements related to the innate immune response in the synovial membrane increased gradually with the modeling prolonged, but decreased significantly after EA treatment. Additionally, the expression of some components in EA1 group was significantly lower than that in EA2 group. These results confirm that synovial inflammation gradually aggravated with time from the early to mid-stage of KOA. EA alleviated the inflammation and histological changes in KOA rabbits by inhibiting the TLRs-mediated innate synovial immune response. This suggests that using EA in the early stage of KOA may achieve a desirable efficacy.

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Mechanism of Salviae Miltiorrhizae Radix et Rhizoma in the treatment of knee osteoarthritis based on network pharmacology

10.21203/rs.3.rs-75437/v1 ◽

2020 ◽

Author(s):

Xiaoqing Shi ◽

Haosheng Zhang ◽

Yue Hu ◽

Xiaochen Li ◽

Songjiang Yin ◽

...

Keyword(s):

Molecular Docking ◽

Knee Osteoarthritis ◽

Molecular Mechanism ◽

Tanshinone Iia ◽

Inflammatory Factors ◽

Venn Diagram ◽

Network Pharmacology ◽

Bioinformatics Analyses ◽

Salviae Miltiorrhizae ◽

Salviae Miltiorrhizae Radix

Abstract Objective: The molecular mechanism of Salviae Miltiorrhizae Radix et Rhizoma (SMRR) in the treatment of knee osteoarthritis (KOA) was analyzed based on network pharmacology.Methods: Active components and potential targets of SMRR were obtained from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP). KOA targets were obtained from the OMIM, DisGeNET, DrugBank, PharmGKB and GeneCards Databases. The potential targets of SMRR in the treatment of KOA were identified by Venn diagram. A protein-protein interaction network was generated with the STRING database. Visualization of the interactions in a potential pharmacodynamic component-target network was accomplished with Cytoscape software. The DAVID database and R software were used for Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway annotation analyses of common targets. Molecular docking of the potential leading components, as determined by efficacy with the core target molecules, was performed with Discovery Studio.Results: Fifty-seven potential pharmacodynamic components and 58 potential targets of SMRR in the treatment of KOA were found. Bioinformatics analyses showed that the IL-17, HIF-1 and TNF signaling pathways, as well as the AGE-RAGE signaling pathway in cases of diabetic complications, are related to the molecular mechanism of SMRR in the treatment of KOA. Molecular docking results showed that luteolin, Tanshinone IIA, Cryptotanshinone and other components of SMRR had strong affinity for MYC, STAT3, CASP3, JUN, CCND1, PTGS2, EGFR, MAPK1, AKT1, VEGFA and other targets.Conclusion: SMRR indirectly regulates IL-17, HIF-1, TNF and other signal transduction pathways by regulating the expression of proteins including PTGS2, MAPK1, EGFR and CASP3, thus playing a role in promoting chondrocyte proliferation, improving microcirculation, eliminating free radicals, and inhibiting inflammatory factors.

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