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18 May 2020 : Laboratory Research  

MiR-423-5p Regulates Cells Apoptosis and Extracellular Matrix Degradation via Nucleotide-Binding, Leucine-Rich Repeat Containing X1 (NLRX1) in Interleukin 1 beta (IL-1β)-Induced Human Nucleus Pulposus Cells

Hanrong Xu1ABCDE, Liefeng Ji1ACDG*, Chunhua Yu1BCDE, Qiming Chen1BCDF, Qiangqiang Ge1BCDF, Yinjiang Lu1BCDF

DOI: 10.12659/MSM.922497

Med Sci Monit 2020; 26:e922497

Abstract

BACKGROUND: Disc degeneration is characterized partly by the degradation in the extracellular matrix (ECM) and excess apoptosis of nucleus pulposus (NP) cells. NLRX1 (nucleotide-binding, leucine-rich repeat containing X1) is different from the other nucleotide-binding-domain and leucine-rich-repeat proteins and mainly located to the mitochondrial. It negatively regulates NF-κB (nuclear factor kappa B) and apoptosis inhibition. However, how NLRX1 is regulated and exerts effects in disc degeneration is unclear. Thus, the study aimed to analyze the effects of NLRX1 on NP cells.

MATERIAL AND METHODS: NLRX1 expression was detected in interleukin (IL)-1β-induced NP cells by western blot and quantitative real-time polymerase chain reaction (qRT-PCR). Then, NLRX1 was overexpressed in IL-1β-induced NP cells to detect apoptosis-related proteins and the extracellular matrix (ECM) by western blot, along with the detection of apoptosis levels using flow cytometry. StarBase predicted miR-423-5p target 3’UTR of NLRX1. Dual luciferase reporter assay showed that miR-423-5p could bind to the 3’UTR of NLRX1. Besides, miR-423-5p significantly affected NLRX1 levels detected by qRT-qPCR.

RESULTS: The miR-423-5p overexpression markedly, and negatively regulated the protective effects of NLRX1 on IL-1β induced NP cells. Thus, our results suggested that miR-423-5p mediated the regulation of NLRX1 to affect apoptosis and ECM levels in IL-1β induced NP cells.

CONCLUSIONS: miR-423-5p and NLRX1 could be potential therapeutic targets for patients with disc degeneration.

Keywords: Extracellular Matrix, intervertebral disc degeneration, Leucine, Mitochondrial Proteins, nucleus pulposus

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750