NOX4 Regulates NLRP3 by Inhibiting the Ubiquitination of LRRC8A to Promote Ferroptosis in Nucleus Pulposus Cells

  • Inflammation. 2025 Oct;48(5):3111-3129. doi: 10.1007/s10753-025-02253-0.
Feng Zhang  #  1  2 ,  Di Cui  #  3 ,  Zhaodong Wang  #  4  5 ,  Yifei Li  1  2 ,  Kangkang Wang  1  2 ,  Haitao Lu  1  2 ,  Haiyang Yu  6  7 ,  Wei Jiao  8  9 ,  Xilong Cui  10  11
Affiliations
  • 1. Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
  • 2. Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China.
  • 3. Medical School of Fuyang, Normal University, No. 100, Qinghe West Road, Yingzhou District, Fuyang, 236000, Anhui, China.
  • 4. Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical University, No.2600, Donghai Dadao, Bengbu, 233000, Anhui, China.
  • 5. Department of Orthopedics, the First Affiliated Hospital of Bengbu Medical University, No. 287,Changhuai Road, Bengbu, 233000, Anhui, China.
  • 6. Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • 7. Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • 8. Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • 9. Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • 10. Department of Orthopedics, Fuyang City People's Hospital, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • 11. Clinical Research Center for Spinal Deformity of Anhui Province, No. 501, Sanqing Road, Fuyang, 236000, Anhui, China. [email protected].
  • # Contributed equally.
PMID: 39909992 DOI: 10.1007/s10753-025-02253-0
Abstract

Intervertebral disc degeneration (IDD) is a significant contributor to low back pain, imposing a considerable socioeconomic burden. Ferroptosis, a novel form of cell death driven by iron and characterized by the accumulation of Reactive Oxygen Species (ROS), has been associated with the progression of IDD. Nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) has been widely recognized as a pivotal factor promoting Ferroptosis across various diseases; however, its precise role in the pathogenesis of IDD remains incompletely understood. Our experimental findings demonstrated a marked upregulation of NOX4 in degenerated cells, accompanied by elevated ROS levels and a diminished mitochondrial membrane potential, indicating the participation of Ferroptosis. Furthermore, the expression of the critical regulatory factor GPX4 was reduced, while ACSL4 levels were significantly increased, further corroborating the involvement of Ferroptosis. Functional loss and gain experiments revealed that NOX4 overexpression augmented Ferroptosis and ROS production while promoting the secretion of inflammatory cytokines. Subsequent studies indicated that the knockdown of NOX4 could reverse tert-butyl hydroperoxide (TBHP)-induced Ferroptosis. Mass spectrometry analysis identified leucine-rich repeat-containing 8A (LRRC8A) as an interacting protein of NOX4, and further validation confirmed that they co-regulate NOD-like Receptor pyrin domain-3 (NLRP3) activation through their interaction. Utilizing a rat model of intervertebral disc degeneration, we further corroborated the role of NOX4 in IDD. This study provides theoretical support for the potential application of NOX4-targeting drugs in the treatment of IDD.

Keywords
Ferroptosis; Intervertebral disc degeneration; NOX4; Reactive oxygen species.