Hydrostatic Pressure Regulates Oxidative Stress through microRNA in Human Osteoarthritic Chondrocytes.

Abstract

Hydrostatic pressure (HP) modulates chondrocytes metabolism, however, its ability to regulate oxidative stress and microRNAs (miRNA) has not been clarified. The aim of this study was to investigate the role of miR-34a, miR-146a, and miR-181a as possible mediators of HP effects on oxidative stress in human osteoarthritis (OA) chondrocytes. Chondrocytes were exposed to cyclic low HP (1-5 MPa) and continuous static HP (10 MPa) for 3 hrs. Metalloproteinases (MMPs), disintegrin and metalloproteinase with thrombospondin motif (ADAMTS)-5, type II collagen (Col2a1), miR-34a, miR-146a, miR-181a, antioxidant enzymes, and B-cell lymphoma 2 (BCL2) were evaluated by quantitative real-time polymerase chain reaction qRT-PCR, apoptosis and reactive oxygen species ROS production by cytometry, and β-catenin by immunofluorescence. The relationship among HP, the studied miRNA, and oxidative stress was assessed by transfection with miRNA specific inhibitors. Low cyclical HP significantly reduced apoptosis, the gene expression of MMP-13, ADAMTS5, miRNA, the production of superoxide anion, and mRNA levels of antioxidant enzymes. Conversely, an increased Col2a1 and BCL2 genes was observed. β-catenin protein expression was reduced in cells exposed to HP 1-5 MPa. Opposite results were obtained following continuous static HP application. Finally, miRNA silencing enhanced low HP and suppressed continuous HP-induced effects. Our data suggest miRNA as one of the mechanisms by which HP regulates chondrocyte metabolism and oxidative stress, via Wnt/β-catenin pathway.

• Authors: Sara Cheleschi, Marcella Barbarino, Ines Gallo, Sara Tenti, Maria Bottaro, Elena Frati, Stefano Giannotti, Antonella Fioravanti
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 21 Issue 10 (May 21 2020) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 32455798 (Publication Type: Journal Article)

Chemical References

• BCL2 protein, human
• COL2A1 protein, human
• Collagen Type II
• MicroRNAs
• Proto-Oncogene Proteins c-bcl-2
• beta Catenin
• ADAMTS5 Protein
• ADAMTS5 protein, human
• MMP13 protein, human
• Matrix Metalloproteinase 13

Topics

• ADAMTS5 Protein (genetics, metabolism)
• Aged
• Apoptosis
• Cells, Cultured
• Chondrocytes (metabolism)
• Collagen Type II (genetics, metabolism)
• Female
• Humans
• Hydrostatic Pressure
• Male
• Matrix Metalloproteinase 13 (genetics, metabolism)
• MicroRNAs (genetics, metabolism)
• Osteoarthritis (genetics, metabolism)
• Oxidative Stress
• Proto-Oncogene Proteins c-bcl-2 (genetics, metabolism)
• Wnt Signaling Pathway
• beta Catenin (genetics, metabolism)