miR‑489‑3p overexpression inhibits lipopolysaccharide‑induced nucleus pulposus cell apoptosis, inflammation and extracellular matrix degradation via targeting Toll‑like receptor 4

Abstract

Intervertebral disc degeneration (IDD) is a common disease with a high morbidity rate, which results in a significant deterioration in the quality of life of patients. MicroRNAs (miRNAs/miRs) are a class of endogenous small non‑coding RNAs that influence target genes and serve critical roles in numerous biological processes. However, the role of miR‑489‑3p in lumbar disc degeneration is yet to be elucidated. In the present study, human NP cells were treated with 10 ng/ml lipopolysaccharide (LPS) for 24 h to investigate the role of miR‑489‑3p in IDD in an <em>in vitro</em> model. Reverse transcription‑quantitative (RT‑q)PCR was performed to determine the expression levels of miR‑489‑3p. Then, the TargetScan database was used to predict the potential binding sites between miR‑489‑3p and Toll‑like receptor (TLR)4, and a dual‑luciferase reporter assay was performed to verify the findings. Subsequently, RT‑qPCR and western blotting were used to analyze the expression levels of TLR4. In addition, human nucleus pulposus (NP) cells were transfected with a miR‑489‑3p mimic and TLR4 overexpression plasmid to study the effects of miR‑489‑3p on LPS‑induced human NP cells. Cell apoptosis and cell viability were also determined using flow cytometry and MTT assays, respectively. Finally, ELISAs were performed to analyze the levels of inflammatory factors. The expression levels of miR‑489‑3p were discovered to be downregulated in LPS‑treated human NP cells. In addition, TLR4 was revealed to be a direct target gene of miR‑489‑3p, and its expression levels were upregulated in LPS‑treated human NP cells. miR‑489‑3p was found to inhibit the LPS‑induced decreases in cell viability and increases in apoptosis, and the concentration of inflammatory cytokines. Furthermore, miR‑489‑3p suppressed the LPS‑induced decreases in extracellular matrix deposition via decreasing the expression levels of aggrecan and collagen type II in human NP cells. Finally, the results revealed that miR‑489‑3p inhibited the LPS‑induced activation of the NF‑κB signaling pathway in human NP cells. Conversely, all of the effects of miR‑489‑3p on LPS‑induced human NP cells were reversed by the TLR4 overexpression plasmid. These findings suggested that miR‑489‑3p may represent a novel therapeutic target for the treatment of IDD.