Transcriptome and alternative splicing analysis of nucleus pulposus cells in response to high oxygen tension: Involvement of high oxygen tension in the pathogenesis of intervertebral disc degeneration

Abstract

High oxygen tension caused by neovascularization in the microenvironment of intervertebral discs (IVDs) is associated with the pathogenesis of IVD degeneration (IDD). Pre-mRNAs undergo alternative splicing (AS) to produce structurally and functionally diverse mRNA and proteins. However, the precise role of high oxygen tension in IDD and the relationship between AS and high oxygen tension in disc cells remain unknown. To investigate the effect of high oxygen tension on disc cells, Affymetrix Rat Transcriptome Array 1.0 was used to determine differentially expressed genes (DEGs) and alternative splicing genes (ASGs) in rat nucleus pulposus (NP) cells treated with 20% O2. NP cells at 1% O2 served as the control. PCR was used for validation. GO and KEGG pathway analysis was performed. Furthermore, the reactive oxygen species (ROS) production, growth, cell cycle and matrix metabolism of NP cells were also investigated. In total, 2499 DEGs and 8451 ASGs were identified. Various GO terms and KEGG pathways were potently associated with IDD, including autophagy, mTOR signaling pathway and angiogenesis. Especially, high oxygen tension increased ROS production in NP cells. It also accelerated the matrix metabolism of NP cells and induced NP cell cycle arrest to retard cell growth. This study, for the first time, analyzes the transcriptome and AS of NP cells in response to high oxygen tension, indicating that high oxygen tension is involved in the establishment and progression of IDD through its wide effects on the viability and function of disc cells.