Endoplasmic reticulum stress‐mediated autophagy alleviates lipopolysaccharide‐induced nucleus pulposus cell pyroptosis by inhibiting CHOP signaling in vitro

• Published in: Journal of biochemical and molecular toxicology 2024-01, Vol.38 (1), p.e23523-n/a
• Main Authors: Chen, Lu, Zhu, Lei, Shi, Hang, Xie, Zhi‐Yang, Jiang, Zan‐Li, Xu, Zheng‐Yuan, Zhang, Zi‐Jian, Wu, Xiao‐Tao
• Format: Article
• Language: English
• Subjects: Assaying; Autophagy; Caspase; CCAAT/enhancer-binding protein; Cell viability; Degeneration; Endoplasmic reticulum; endoplasmic reticulum stress; Homeostasis; Immunofluorescence; Inflammation; intervertebral disc degeneration; Intervertebral discs; Lipopolysaccharides; Molecular modelling; Necrosis; Nuclei (cytology); Nucleus pulposus; nucleus pulposus cell; Preconditioning; Proteins; Pyroptosis; Regulatory mechanisms (biology); Signal transduction
• ISSN: 1095-6670; 1099-0461
• DOI: 10.1002/jbt.23523

Pyroptosis, a newly discovered pro‐inflammatory programmed necrosis of cells, serves as an initiating and promoting event that leads to intervertebral disc (IVD) degeneration (IDD). Endoplasmic reticulum stress (ERS) and autophagy are vital regulatory mechanisms of cellular homeostasis, which is also closely related to IDD. However, the role and relationship of ERS and autophagy in the pyroptosis of nucleus pulposus cell (NPC) are not well understood. In this research, we aimed to elucidate the role and mechanism of ERS‐C/EBP homologous protein (CHOP) in lipopolysaccharide (LPS)‐induced cell pyroptosis and determine its interaction with autophagy. ERS and autophagy inducers or inhibitors were used or not in the preconditioning of rat NPCs. Cell viability, pyroptosis‐related protein expression, caspase‐1 activity assay, and enzyme‐linked immunosorbent assay were performed to observe rat NPC pyroptosis after the treatment of LPS. Activation of the ERS pathway and autophagy were assessed by quantitative real‐time PCR, western blot analyses, and immunofluorescence staining assay to classify the molecular mechanisms. Our results showed that LPS stimulation induced NPC pyroptosis with concomitant activation of the ERS‐CHOP pathway and initiated autophagy. Activation of the ERS‐CHOP pathway exacerbated rat NPC pyroptosis, whereas autophagy inhibited cell pyroptosis. LPS‐induced cell pyroptosis and CHOP upregulation were negatively regulated by autophagy. LPS‐induced autophagy was depressed by the ERS inhibitor but aggravated by the ERS inducer. Taken together, our findings suggested that LPS induced NPC pyroptosis by activating ERS‐CHOP signaling and ERS mediated LPS‐induced autophagy, which in turn alleviated NPC pyroptosis by inhibiting CHOP signaling.