cyclin-d1 and Osteoarthritis--Knee

Knee osteoarthritis (KOA) is one of the common age-degenerative diseases. Recent studies have demonstrated that the pathogenesis of KOA is closely related to synovial lesions. Jiawei Duhuo Jisheng mixture (JDJM) has shown great potential in the treatment of KOA. However, the effect and mechanism of JDJM on synovial lesions of KOA remain unclear.. The regulatory effect of JDJM on the Wnt/β-catenin signaling pathway in KOA inflamed synovium was studied via in vitro and in vivo experiments, respectively.. For the in vitro experiment, fibroblasts were isolated from the rabbit synovium with KOA. The fibroblasts were grouped as follows: the vehicle group was given 0.5% FBS; the inhibitor group was treated with 0.5% volume fraction of XAV939; the normal serum groups and JDJM serum groups were treated with 5%, 10%, and 20% volume fractions of normal serum and JDJM-containing serum. The expression levels of Wnt3a, β-catenin, Cyclin D1, metalloproteinase-7(MMP-7) and cyclooxygenase-2(COX-2) were detected by different assays 48 and 72 h after the intervention. For the in vivo experiment, the rabbit KOA model was prepared using the improved Hulth modeling method, whereby all rabbits were randomly divided into normal control, model control, positive control, and traditional Chinese medicine (TCM) groups. The expression levels of Wnt3a, β-catenin, Cyclin D1, MMP-7 and COX-2 were detected by different assays in the 2, 4, and 8 weeks of treatment.. In the two test results of in vitro experiments, the normal serum group was compared with the JDJM-containing serum group with the same volume fraction, demonstrating that mRNA transcription and protein expression levels of Wnt3a, β-catenin, Cyclin D1, MMP-7, and COX-2 in the latter decreased (P < 0.05), with more pronounced effects observed in the group treated with 20% volume fraction of JDJM serum. Compared with the inhibitor group, there was no significant difference (P > 0.05) in the mRNA transcription and protein expression levels, i.e., Wnt3a, β-catenin, Cyclin D1, and MMP-7 were observed in the JDJM serum groups, except for a significant decrease (P < 0.05) in the level of mRNA transcription and protein expression of COX-2. Based on the in vivo experiment, compared to the model control group, articular cartilage, synovial hyperplasia, and the inflammatory reaction of the TCM group at different treatment times were significantly improved. The mRNA transcription level of Wnt3a, β-catenin, Cyclin D1, MMP-7 and COX-2 detected by RT-qPCR and the protein expression level of Wnt3a, β-catenin, Cyclin D1, MMP-7 and COX-2 detected by Western blot were significantly reduced (P < 0.05), and the effect was more evident at the eighth week.. JDJM can regulate the synovial Wnt/β-catenin signaling pathway in KOA models, reduce the mRNA transcription and protein expression levels of Wnt3a, β-catenin, Cyclin D1, MMP-7, and COX-2 in the synovium, thus inhibiting synovial inflammation and protecting articular cartilage, which could be the key mechanism of action in treating this disease.

Topics: Animals; beta Catenin; Cyclin D1; Cyclooxygenase 2; Drugs, Chinese Herbal; Inflammation; Matrix Metalloproteinase 7; Osteoarthritis, Knee; Rabbits; RNA, Messenger; Synovial Membrane; Wnt Signaling Pathway

This study aimed to elucidate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation combined with the administration of Lugua polypeptide injection into the knee joint cavity to treat knee osteoarthritis (KOA) in rabbits.. Sixty white New Zealand rabbits were randomly divided into the blank, model, Lugua polypeptide, BMSC, and combined (Lugua polypeptide plus BMSC) groups, with 12 rabbits in each group. The mRNA and protein expression levels of cyclin D1, bcl-2, TIMP-1, p21, caspase-3, Bax, MMP-1, MMP-13, TLR-4, and NF-κB p65 in chondrocytes, and levels of IL-1, NO, TNF-α, and IL-6 in the synovial fluid were compared.. The severity of cartilage damage in the combined group was significantly less (. BMSC transplantation combined with Lugua polypeptide injection may improve KOA-related cartilage tissue damage in rabbits.

Topics: Animals; bcl-2-Associated X Protein; Bone Marrow; Caspase 3; Chondrocytes; Cyclin D1; Interleukin-1; Interleukin-6; Knee Joint; Matrix Metalloproteinase 1; Matrix Metalloproteinase 13; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; NF-kappa B; Osteoarthritis, Knee; Rabbits; RNA, Messenger; Tissue Inhibitor of Metalloproteinase-1; Toll-Like Receptor 4; Tumor Necrosis Factor-alpha

Recently, accumulating evidence demonstrated that the long non-coding RNAs (lncRNAs) play important roles in osteoarthritis (OA) progression. However, the role of lncRNA FOXD2-AS1 on OA is still unclear. In the present study, qRT-PCR showed that expression of FOXD2-AS1 and Cyclin D1 (CCND1) was upregulated in OA cartilage tissues, while miR-206 expression was significantly decreased. CCK-8 and colony formation assays showed that FOXD2-AS1 could promote chondrocytes viability. Flow cytometry analysis showed that FOXD2-AS1 inhibition arrested chondrocytes in G0/G1 phase and induced cells apoptosis. Furthermore, luciferase reporter assay and RIP assay showed that FOXD2-AS1 could function as a sponge of miR-206. Rescue assays showed that miR-206 inhibitors reversed the effects of FOXD2-AS1 suppression on chondrocytes viability. In addition, we identified that CCND1 acted as a direct target of miR-206. FOXD2-AS1 suppression could inhibit CCND1 expression in chondrocytes, while miR-206 inhibitors reversed CCND1 expression. Moreover, rescue assays indicated that CCND1 overexpression reversed the effects of FOXD2-AS1 suppression on chondrocytes viability. Taken together, these data indicated that FOXD2-AS1 could promote the growth of chondrocytes by targeting miR-206/CCND1 axis.

Topics: Apoptosis; Cell Cycle Checkpoints; Cell Proliferation; Cell Survival; Cells, Cultured; Chondrocytes; Cyclin D1; Female; Gene Expression Regulation; Humans; Male; MicroRNAs; Middle Aged; Osteoarthritis, Knee; RNA, Long Noncoding; Signal Transduction; Time Factors

The aim of this study was to investigate the roles of cyclin D1, CDK4, and p53 in knee osteoarthritis (KOA).. A total of 76 healthy controls and 154 KOA cases (grades ranging from II to IV) were recruited. Protein expression of cyclin D1, CDK4, and p53 were detected by immunohistochemistry, and mRNA expression levels of the cyclin D1, the CDK4, and the p53 genes were measured by reverse transcription-polymerase chain reaction.. Both protein and mRNA expression levels of cyclin D1 and CDK4 were significantly lower in KOA cases than those in healthy controls, while protein and mRNA expression of p53 was significantly higher in KOA cases than that in healthy controls (all p < 0.05). As the grades of KOA increased, Cyclin D1 and CDK4 mRNA expressions decreased, whereas p53 mRNA expression increased (all p < 0.05). In KOA cases, mRNA expression of Cyclin D1 was positively correlated to CDK4 mRNA levels (r = 0.386, p < 0.001), while negatively correlated with p53 mRNA levels (r = -0.227, p = 0.005).. Expression of the Cyclin D1, CDK4, and p53 genes are correlated with the disease grades of KOA.

Topics: Adult; Aged; Case-Control Studies; Cell Cycle Proteins; Cyclin D1; Cyclin-Dependent Kinase 4; Female; Humans; Immunohistochemistry; Male; Middle Aged; Osteoarthritis, Knee; RNA, Messenger; Tumor Suppressor Protein p53