MicroRNA‑204‑5p inhibits the osteogenic differentiation of ankylosing spondylitis fibroblasts by regulating the Notch2 signaling pathway

Abstract

ankylosing spondylitis (aS) is a chronic inflammatory systemic disease and is difficult to detect in the early stages. The present study aimed to investigate the role of microRNA (miR)-204-5p in osteogenic differentiation of AS fibroblasts. Bone morphogenetic protein 2 (BMP-2) was used to induce osteogenic differentiation. Cells were divided into the following groups: AS group, AS + BMP-2 group, AS + BMP-2 + miR-negative control group, AS + BMP-2 + miR-204-5p mimics group and AS + BMP-2 + miR-204-5p mimics + pcDNA-Notch2 group. The expression levels of miR-204-5p, Notch2, runt-related transcription factor 2 (RUNX2) and osteocalcin were detected via reverse transcription-quantitative PCR analysis. The binding site between Notch2 and miR-204-5p was predicted using TargetScan software and verified via the dual-luciferase reporter assay. Alkaline phosphatase (ALP) activity was assessed via the ALP assay, while the mineralized nodules area was determined via the Alizarin Red S staining assay. The results demonstrated that Notch2 is a target gene of miR-204-5p. Furthermore, treatment with BMP-2 significantly decreased miR-204-5p expression, and significantly increased ALP activity, the mineralized nodules area and the expression levels of Notch2, RUNX2 and osteocalcin in ligament fibroblasts (all P<0.05). Conversely, transfection with miR-204-5p mimics significantly increased miR-204-5p expression, and significantly decreased ALP activity, the mineralized nodules area and the expression levels of Notch2, RUNX2 and osteocalcin in ligament fibroblasts (all P<0.05). Notably, transfection with pcDNA-Notch2 significantly reversed the inhibitory effects induced by miR-204-5p mimics on the osteogenic differentiation of ligament fibroblasts (all P<0.05). Furthermore, miR-204-5p inhibited the osteogenic differentiation of ligament fibroblasts in patients with AS by targeting Notch2. Thus, miR-204-5p may negatively regulate Notch2 expression and may be a potential therapeutic target for AS. Collectively, the results of the present study provide a theoretical basis for the effective treatment of patients with AS.

Involvement of MiRNA-211-5p and Arhgap11a Interaction During Osteogenic Differentiation of MC3T3-E1 Cells

“…For example, Yu et al found that, in calcific aortic valve disease, increased expression of miR-204-5p inhibited Runx2 expression at the post-transcriptional level, resulting in suppression of osteogenic differentiation finally (31). Another study on ankylosing spondylitis presented by Zhao et al also indicated decreased miR-204-5p expression and increased activity of ALP, as well as elevated expression of RUNX2 and OCN, contributed to enhancing osteogenic differentiation of ankylosing spondylitis fibroblasts (32). Wu et al demonstrated lncRNA LEF1-AS1 favored osteogenic differentiation of dental pulp stem cells via repressing the expression of miR-24-3p (33).…”

miR‑148a‑3p facilitates osteogenic differentiation of fibroblasts in ankylosing spondylitis by activating the Wnt pathway and targeting DKK1

“…In the present study, an elevated miR-148a-3p expression was identified in the AS ligament tissues. Osteogenic differentiation of fibroblasts is the primary cause of osteophyte formation and ankylosis in AS ( 31 ). Previous results validated the functionality of vimentin fragments as potential markers of rheumatoid synovial fibroblasts ( 32 ).…”

The Potential Role of Genetics, Environmental Factors, and Gut Dysbiosis in the Aberrant Non-Coding RNA Expression to Mediate Inflammation and Osteoclastogenic/Osteogenic Differentiation in Ankylosing Spondylitis

“…During osteogenic differentiation, miR-124, β-catenin, osteorix and RUNX2 expression gradually increased, while that of GSK-3β gradually declined. Zhao et al (2020) have demonstrated that miR-204-5p can negatively regulate NOTCH 2 expression in osteogenic differentiation in the ligament fibroblasts derived from AS patients in vitro . These results may provide a therapeutic basis for the effective treatment for patients with AS.…”