Inhibition of miR-33b-5p promotes osteoblast differentiation by regulating ABCA1 in cellular and animal models of osteoporosis
Abstract
INTRODUCTION: Osteoporosis is a major health risk for postmenopausal women and is primarily driven by estrogen decline-induced bone loss. As critical post-transcriptional regulators of gene expression, dysregulated microRNAs (miRs) participate in the pathogenesis of osteoporosis by disrupting key signaling pathways that govern bone homeostasis and remodeling. This study investigated the functional role of the miR-33b-5p/ABCA1 axis in cellular and murine models of osteoporosis. MATERIAL AND METHODS: Alizarin Red S staining and alkaline phosphatase (ALP) activity assays were performed to detect osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). ABCA1 mRNA and protein levels, as well as osteogenic proteins such as RUNX2, OPN, and OCN, were quantified by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. Dexamethasone (DEX)-stimulated BMSCs were employed as an in vitro osteoporosis model. To overexpress ABCA1 in BMSCs, the cells were transfected with pcDNA3.1 vectors carrying the full-length ABCA1 coding sequence. ABCA1 silencing was accomplished using short hairpin RNA targeting ABCA1 (sh-ABCA1), whereas miR-33b-5p inhibition was achieved using a specific miR-33b-5p inhibitor. The direct interaction between miR-33b-5p and its downstream target ABCA1 was validated using luciferase reporter assays. After an ovariectomized (OVX) mouse model was established, micro-CT was performed to detect bone density…
Read the paper
Related studies
- Irisin Enhances Osteoblast DifferentiationIn Vitro
- Glucocorticoids suppress Wnt16 expression in osteoblasts in vitro and in vivo
- Interaction between Schwann Cells and Osteoblasts In Vitro
- Administration of cinnamaldehyde promotes osteogenesis in ovariectomized rats and differentiation of osteoblast in vitro
- The Effect of the Major Components of Fructus Cnidii on Osteoblasts In Vitro
- Estrogen depletion alters osteogenic differentiation and matrix production by osteoblasts in vitro
Go deeper in Evidence Hub
Evidence Hub adds a study data sheet (design, sample size, evidence level, key numbers), an AI summary and a chat grounded in the paper's text.