circSLTM knockdown attenuates chondrocyte inflammation, apoptosis and ECM degradation in osteoarthritis by regulating the miR-515-5p/VAPB axis.

Osteoarthritis (OA) is a prevalent joint disorder characterized by cartilage degeneration. Circular RNAs (circRNAs) have emerged as pivotal players in OA progression, orchestrating various biological processes such as proliferation, apoptosis, inflammation, and extracellular matrix (ECM) reorganization. Among these circRNAs, circSLTM exhibits aberrant expression in OA, yet its precise regulatory mechanism remains elusive. This study aimed to elucidate the regulatory mechanisms of circSLTM in OA pathogenesis, with a focus on its role as a competing endogenous RNA (ceRNA). Human cartilage tissues were procured from both OA patients and non-OA individuals, while human chondrocyte cells were subjected to lipopolysaccharide (LPS) treatment to mimic OA-like conditions. Our findings revealed upregulation of circSLTM in OA patients and LPS-treated chondrocytes. Loss-of-function assays were conducted, demonstrating that silencing circSLTM via shRNAs mitigated LPS-induced effects on chondrocytes, as evidenced by enhanced proliferation, reduced apoptosis, and inflammatory factors, and altered expression of extracellular matrix proteins. Further exploration into the regulatory mechanism of circSLTM unveiled its interaction with microRNA-515-5p (miR-515-5p) to modulate vesicle-associated membrane protein (VAPB) expression in chondrocytes. VAPB, also upregulated in OA, was positively regulated by circSLTM. Rescue assays corroborated that VAPB overexpression reinstated the protective effects of circSLTM knockdown on LPS-treated chondrocytes. Moreover, concurrent knockdown of both circSLTM and VAPB demonstrated synergistic protection against LPS-induced chondrocyte injury. Additionally, we delineated that LPS triggered the activation of the NF-κB pathway in chondrocytes, which was counteracted by circSLTM silencing. To assess the effects of circSLTM on OA in vivo, anterior cruciate ligament transection (ACLT) mouse models were established, revealing that circSLTM deficiency ameliorated cartilage defects in vivo. In conclusion, circSLTM exacerbates osteoarthritis progression by orchestrating the miR-515-5p/VAPB axis and activating the NF-κB pathway, providing novel insights for targeted therapy in OA management.

The effects of Omarigliptin on promoting osteoblastic differentiation.

Osteoporosis significantly impacts the normal life of the elderly and is reported to be closely related to dysfunction of osteoblastic differentiation. Runt-related transcription factor-2 (Runx2) is a critical transcriptional factor involved in the regulation of osteoblast differentiation. Omarigliptin is a novel dipeptidyl peptidase-4 (DDP-4) inhibitor and this study proposes to probe into its possible therapeutic function against Osteoporosis by investigating its impacts on osteoblastic differentiation. Osteogenic medium was used to induce osteoblastic differentiation in MC3T3­E1 cells, and was verified by the increased alkaline phosphatase (ALP) activity, enhanced mineralization, and promoted expression level of osteoblastic differentiation-related factors, including bone morphogenetic protein-2 (BMP-2), ALP, osteocalcin (Ocn), collagen type I alpha 1 (Col1a1), Collagen Type I alpha 2 (Col1a2), Runx2, osterix (Sp7), fibroblast growth factor receptor 2 (Fgfr2), and fibroblast growth factor receptor 3 (Fgfr3), accompanied by the activation of the p38 and Akt pathways. After treatment with Omarigliptin, the ALP activity and mineralization were further promoted, accompanied by the further upregulation of osteoblastic differentiation-related factors, and activation of the p38 and Akt pathways. Lastly, Omarigliptin-induced osteoblastic differentiation, promoted ALP activity, and increased expression levels of Sp7, Fgfr2, Fgfr3, BMP-2, Ocn, ALP, Col1a1, and Col1a2, in the osteogenic medium- cultured MC3T3­E1 cells were dramatically abolished by the knockdown of Runx2. Taken together, our data reveal that Omarigliptin promoted osteoblastic differentiation by regulating Runx2.

MiRNA-19a-3p alleviates the progression of osteoporosis by targeting HDAC4 to promote the osteogenic differentiation of hMSCs.

To clarify the function of microRNA-19a-3p (miRNA-19a-3p) in the osteogenic differentiation of human-derived mesenchymal stem cells (hMSCs) and the potential mechanism. Serum levels of miRNA-19a-3p, RUNX2 and OCN in osteoporosis patients and controls were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Alkaline phosphatase (ALP) content and calcification ability during the process of osteogenic differentiation were examined by ALP staining and alizarin red staining, respectively. After altering miRNA-19a-3p level by transfection of miRNA-19a-3p mimic or inhibitor, we detected relative levels of miRNA-19a-3p, RUNX2 and OCN in hMSCs by qRT-PCR. The binding relationship between miRNA-19a-3p and HDAC4 was predicted by TargetScan and further verified by dual-luciferase reporter gene assay. Relative expression of HDAC4 was detected by Western blot and qRT-PCR in hMSCs transfected with miRNA-19a-3p mimic or inhibitor. Regulatory effects of miRNA-19a-3p/HDAC4 axis on osteogenic differentiation of hMSCs were evaluated. MiRNA-19a-3p was downregulated in osteoporosis patients. Its level gradually increased in hMSCs with the prolongation of osteogenic differentiation. Overexpression of miRNA-19a-3p upregulated levels of RUNX2 and OCN, and enhanced ALP activity. Knockdown of miRNA-19a-3p obtained the opposite trends. Dual-luciferase reporter gene assay verified that miRNA-19a-3p could target to 3'UTR of HDAC4. Protein level of HDAC4 was negatively regulated by miRNA-19a-3p in hMSCs. More importantly, co-overexpression of miRNA-19a-3p and HDAC4 could reverse the regulatory effects of miRNA-19a-3p on enhancing ALP activity and upregulating RUNX2 and OCN. MiRNA-19a-3p promotes the osteogenic differentiation of hMSCs by inhibiting HDAC4 expression, thus alleviating the progression of osteoporosis.