MiR-494-3p knockdown promotes osteogenic differentiation of bone marrow mesenchymal stem cells in ovariectomised rats by targeting Sirt1/TLR4/NF-κB pathway
INTRODUCTION: Osteoporosis is a bone disease characterised by low bone mass, deterioration of bone tissue, and disruption of bone microarchitecture. MicroRNAs have been found to play an important role in osteoporosis. MicroRNA (miR)-494 is inhibited during bone angiogenesis, and its overexpression r...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Via Medica
2025-06-01
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| Series: | Folia Histochemica et Cytobiologica |
| Subjects: | |
| Online Access: | https://journals.viamedica.pl/folia_histochemica_cytobiologica/article/view/104450 |
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| Summary: | INTRODUCTION: Osteoporosis is a bone disease characterised by low bone mass, deterioration of bone tissue, and disruption of bone microarchitecture. MicroRNAs have been found to play an important role in osteoporosis. MicroRNA (miR)-494 is inhibited during bone angiogenesis, and its overexpression reduces osteogenic differentiation gene expression. Sirtuin 1 (Sirt1) is a histone deacetylase with multiple cellular activities including increasing bone mass and delaying the onset of osteoporosis. MiR-494-3p has been predicted in computer-assisted bioinformatics analysis to target the 3’UTR of Sirt1 mRNA. The aim of the present study was to assess the effect of miR-494-3p on ovariectomy (OVX)-induced osteoporosis in rats and the relevant mechanisms. MATERIAL AND METHODS: Osteoporosis in female rats was induced by OVX, and bone microarchitectural changes were evaluated by means of microCT. MiR-494-3p and Sirt1 expression in femurs were evaluated by RT-qPCR or Western blotting. Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat femurs and identified by flow cytometry. Then, BMSCs were transfected with miR-494-3p inhibitor/mimic, si-Sirt1 and negative controls as well as pcDNA3.1-TLR4 and empty pcDNA3.1 vector. Osteogenic cell differentiation was assessed via Alizarin Red, alkaline phosphatase (ALP) and Oil Red O staining. RESULTS: MiR-494-3p level was upregulated, and Sirt1 mRNA and protein levels were downregulated, in femurs of OVX rats. Functionally, miR-494-3p inhibited osteogenic differentiation of cultured rat BMSCs. Mechanistically, miR-494-3p regulated the Sirt1 3’UTR to activate TLR4/NF-κB signalling, and Sirt1 inhibition and TLR4 overexpression reversed the enhancing effect of miR-494-3p knockdown on osteogenic differentiation. CONCLUSIONS: MiR-494-3p represses osteogenic differentiation of BMSCs in OVX rats through Sirt1/TLR4/NF-κB signalling. |
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| ISSN: | 0239-8508 1897-5631 |