MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes.

Sara Cheleschi,Sara Tenti,Ines Gallo,Claudio Corallo,Stefano Giannotti,Nicola Mondanelli,Antonio Giordano,Antonella Fioravanti,Marcella Barbarino

doi:10.3390/cells8080874

Abstract

Current evidence suggests a complex interaction between adipokines and microRNA (miRNA) in osteoarthritis (OA) pathogenesis. The present study explored the role of miR-34a and miR-181a in regulating apoptosis and oxidative stress induced by visfatin in human OA chondrocytes. Chondrocytes were transfected with miR-34a and miR-181a inhibitors and stimulated with visfatin for 24 h, in the presence of nuclear factor (NF)-κB inhibitor (BAY-11-7082, 2 h pre-incubation). Apoptosis and reactive oxygen species (ROS) production were detected by cytometry, miRNA, antioxidant enzymes, nuclear factor erythroid (NRF)2 and B-cell lymphoma (BCL)2 expressions by quantitative real time polymerase chain reaction (real time PCR) and western blot. P50 NF-κB subunit was measured by immunofluorescence. Visfatin significantly induced apoptosis and superoxide anion production, increased miR-34a, miR-181a, superoxide dismutase (SOD)-2, catalase (CAT), NRF2 and decreased BCL2 gene and protein expression in OA chondrocytes. All the visfatin-caused effects were suppressed by using miR-34a and miR-181a inhibitors. Pre-incubation with BAY-11-7082 counteracted visfatin-induced expression of miRNA, BCL2, SOD-2, CAT and NRF2. Inhibition of miR-34a and miR-181a significantly reduced the activation of p50 NF-κB. Visfatin confirms its ability to induce apoptosis and oxidative stress in human OA chondrocytes; these effects appeared mediated by miR-34a and miR-181a via NF-κB pathway. We highlight the relevance of visfatin as potential therapeutic target for OA treatment.

Highlights

MicroRNA are an abundant class of conserved non-coding C/D Box 25 (RNA) molecules of 22–25 nucleotides which emerged as important post-transcriptional regulators of gene expression either through the enhancement of degradation, by suppressing translation or via other mechanisms [1].Cells 2019, 8, 874; doi:10.3390/cells8080874 www.mdpi.com/journal/cellsThey have been associated with controlling important cellular processes or pathological features [1].Several groups of investigators proposed that altered miRNA expression has been linked to regulation of cartilage homeostasis and osteoarthritis (OA) [2,3]
To confirm whether the expression of miR-34a and miR-181a was induced by visfatin stimulus, miR-34a and miR-181a inhibitors were transfected into OA chondrocytes (Figure 2)
Transfection experiments on our cultures demonstrate the involvement of miR-34a and miR-181a in the modulation of apoptosis and oxidative stress processes induced by visfatin

Summary

Introduction

MicroRNA (miRNA) are an abundant class of conserved non-coding RNA molecules of 22–25 nucleotides which emerged as important post-transcriptional regulators of gene expression either through the enhancement of degradation, by suppressing translation or via other mechanisms [1].Cells 2019, 8, 874; doi:10.3390/cells8080874 www.mdpi.com/journal/cellsThey have been associated with controlling important cellular processes or pathological features [1].Several groups of investigators proposed that altered miRNA expression has been linked to regulation of cartilage homeostasis and osteoarthritis (OA) [2,3]. MicroRNA (miRNA) are an abundant class of conserved non-coding RNA molecules of 22–25 nucleotides which emerged as important post-transcriptional regulators of gene expression either through the enhancement of degradation, by suppressing translation or via other mechanisms [1]. They have been associated with controlling important cellular processes or pathological features [1]. Several groups of investigators proposed that altered miRNA expression has been linked to regulation of cartilage homeostasis and osteoarthritis (OA) [2,3]. One of the possible mechanisms by which miRNA induce articular cartilage damage in OA could be represented by the variation of cellular redox state. Obesity has been traditionally thought of as one of the most influential risk factors for OA [6,7]

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Conclusion