MicroRNA-483-5p Modulates the Expression of Cartilage-Related Genes in Human Chondrocytes through Down-Regulating TGF-<i>β</i>1 Expression

Abstract

Transforming growth factor-β1 (TGF-β1) has been reported to improve chondrocytes phenotype and function. The expression levels of microRNA-483-5p (miR-483-5p), a potential regulator of TGF-β signaling pathway, were elevated in chondrocytes of patients with osteoarthritis. In this study, we aimed to explore the role of miR-483-5p for the expression of cartilage-related genes in chondrocytes. Human chondrocytes were harvested from femoral condyle and tibial plateau of different donors (n = 10) following amputation due to sarcomas not involving the joint space. The expression levels of miR-483-5p and TGF-β1 mRNA were measured by qRT-PCR. Runt-related transcription factor 2 (RUNX2) is a transcription factor involved in chondrocyte differentiation, and type II collagen-degrading matrix metalloproteinase13 (MMP13) contributes to cartilage degradation. qRT-PCR was also used to measure the levels of RUNX2 and MMP-13 mRNAs, as well as type II collagen alpha 1 (COL2A1) and aggrecan mRNAs. COL2A1 and aggrecan are major cartilage extracellular matrix proteins that are essential for normal cartilage function. The expression levels of miR-483-5p were significantly increased in chondrocytes from Passage 0 to 2, whereas the expression levels of TGF-β1 mRNA were marginally decreased. Passage 1 chondrocytes were employed for following experiments. MiR-483-5p overexpression reduced TGF-β1 expression, while miR-483-5p knockdown dramatically elevated TGF-β 1 expression both at mRNA and protein levels. Further, miR-483-5p overexpression significantly decreased the levels of COL2A1 and aggrecan mRNAs, and increased those of RUNX2 and MMP13 mRNAs, by down-regulating TGF-β1 expression. These findings suggest that modulating miR-483-5p expression may contribute to maintaining the cartilage tissues.

MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis

“…Col2a1 (type II collagen alpha 1) and aggrecan are major cartilage extracellular matrix proteins that are essential for normal cartilage function. Runx2 (runt-related transcription factor 2) is a transcription factor involved in chondrocyte differentiation and hypertrophy, and type II collagen-degrading MMP-13 (matrix metalloproteinase-13) contributes to cartilage degradation [18]. To determine the role of MSC-Exos in chondrocytes, we detected the levels of these key cartilage genes with MSC-Exos treatment in OA mice-derived chondrocytes.…”

Onset and Progression of Human Osteoarthritis—Can Growth Factors, Inflammatory Cytokines, or Differential miRNA Expression Concomitantly Induce Proliferation, ECM Degradation, and Inflammation in Articular Cartilage?

“…Also, OA AC micropellet cultures have higher miR-483 level, compared to normal AC micropellet cultures [210]. TGF-β1 is downregulated by overexpression of miR-483 mimic in monolayer cultures from human normal knee AC, which coincides with COL2A1 and ACAN mRNA depletion and RUNX2 and MMP13 upregulation [230]. In human MSPC miR-483 directly targets SMAD4 , which suppresses chondrogenesis.…”

Regulation of microRNA‐221, ‐222, ‐21 and ‐27 in articular cartilage subjected to abnormal compressive forces

“…2015) and catabolic outcomes (Xu et al . 2017; Wang et al . 2019); hence, its observed reduction in response to abnormal load may reflect an attempt at tissue remodelling.…”

“…A reduction in miR-483 levels was observed in response to non-physiological load and is the first report of its mechano-sensitivity in articular cartilage. Its potential role in cartilage homeostasis is not well defined, with conflicting evidence suggesting anabolic (Yang et al 2015) and catabolic outcomes (Xu et al 2017;Wang et al 2019); hence, its observed reduction in response to abnormal load may reflect an attempt at tissue remodelling.…”