MiR-128-3p Post-Transcriptionally Inhibits WISP1 to Suppress Apoptosis and Inflammation in Human Articular Chondrocytes via the PI3K/AKT/NF-κB Signaling Pathway

Abstract

In osteoarthritis (OA), the synthesis and decomposition of the extracellular matrix (ECM) are imbalanced. High expression levels of Wnt1-inducible signaling pathway protein 1 (WISP1) promote the synthesis of matrix metalloproteinases and induce the degradation of cartilage, which aggravates the OA. The aim of this study was to explore the role of miR-128-3p in the development of OA. In the present study, the expression of WISP1 and miR-128-3p in osteoarthritic tissues and chondrocytes was detected using quantitative reverse transcription PCR (RT-qPCR) and Western blotting. Then we predicted that WISP1 might be a potential target gene of miR-128-3p by TargetScan and verified using luciferase reporter gene assay. The effect of miR-128-3p or WISP1 on chondrocytes was evaluated by cell proliferation assay, apoptosis, and caspase-3 activity assay. To further reveal the molecular mechanisms of miR-128-3p in osteoarthritic development, the degradation of chondrocyte matrix and production of proinflammatory cytokines in osteoarthritic chondrocyte model were detected by ELISA. To mimic the osteoarthritic microenvironment in vitro studies, chondrocytes were stimulated with interleukin (IL)-1β, and then we found that the expression of miR-128-3p was downregulated. Overexpression of WISP1 inhibited the proliferation of chondrocytes, which induced apoptosis, degradation of chondrocyte matrix, production of proinflammatory cytokines, and activated the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway. Then, we identified that miR-128-3p was a negative regulator of WISP1 by directly targeting its 3′-untranslated region (UTR). Moreover, the PI3K allosteric activator 740 Y-P abolished the inhibition of miR-128-3p in apoptosis, degradation of chondrocyte matrix, and inflammation. Our results showed that miR-128-3p targets WISP1 to regulate chondrocyte proliferation, apoptosis, degradation of chondrocyte matrix, and production of proinflammatory cytokines via the PI3K/Akt/NF-κB pathway, which plays a suppressed role in OA.

MicroRNA-128-3p suppresses interleukin-1β-stimulated cartilage degradation and chondrocyte apoptosis via targeting zinc finger E-box binding homeobox 1 in osteoarthritis

“…For example, miR-128-3p could directly target SIRT1 to mediate the TNF-α-induced inflammatory response in bone marrow mesenchymal stem cells [ 26 ]. miR-128-3p also attenuated chondrocyte apoptosis, ECM degradation and inflammation via targeting Wnt-induced secreted protein 1 in OA [ 12 ]. Herein, we demonstrated that miR-128-3p was downregulated in chondrocytes and OA tissues.…”

“…miR-145 contributed to impaired extracellular matrix (ECM) degradation in OA cartilage via targeting Smad3 [ 11 ]. Interestingly, miR-128-3p was identified to be downregulated and play a suppressive role in OA [ 12 ]. However, the specific effects of miR-128-3p in the pathogenesis of OA remain unknown.…”

MiR-128-3p inhibits vascular smooth muscle cell proliferation and migration by repressing FOXO4/MMP9 signaling pathway

“…Importantly, miR-128-3p may not only regulate FOXO4 expression through binding its 3′UTR. MiR-128-3p is reported to regulate various pathways / genes, which can probably regulate FOXO4 expression, including Erk signaling, SIRT1, PI3K/AKT pathway and so on (Liu et al 2020b , 2019b ; Wang et al 2014 ; Zhao et al 2019b ; Kobayashi et al 2005 ; Huo et al 2019 ; Chen and Li 2020 ; Gong et al 2019 ; Chang et al 2013 ) (Fig. 7 ).…”

DNA methylation and noncoding RNA in OA: Recent findings and methodological advances

“…For example, knockdown via antisense oligonucleotide of miR-181a-5p attenuates murine OA [ 69 ], whereas miRNA-34a-5p promotes joint destruction in murine models [ 70 ] Two homologous miRNAs, miR-204 and miR-211, work in concert to maintain joint homeostasis and prevent OA development, principally through controlling Runx2 expression [ 71 ]. miRNA-128a worsens murine OA by suppressing chondrocyte autophagy via controlling Atg12 expression [ 72 ] and in human chondrocytes via interactions with WISP1 suppressing apoptosis and inflammation [ 73 ]. miR-29a reduces synovitis in synovium of human knee OA patients and has been shown in vitro to target VEGF [ 74 ].…”