Oxidative damage induces apoptosis and promotes calcification in disc cartilage endplate cell through ROS/MAPK/NF-κB pathway: Implications for disc degeneration.
Cartilage endplate (CEP) cell calcification and apoptosis play a vital role in the intervertebral disc degeneration (IVDD). Oxidative stress is a key factor in inducing programmed cell death and cartilage calcification. However, the cell death and calcification of cartilage endplate cells under oxidative stress have never been described. The present study investigated the apoptosis and calcification in the cartilage endplate cell under oxidative stress induced by H2O2to understand the underlying mechanism of IVDD. The cartilage endplate cells isolated from human lumbar discs were subjected to different concentrations of H2O2for various time periods. The cell viability was determined by CCK-8 assay, whereas Western blot, immunofluorescence, and Alcian blue, Alizarin red, and Von Kossa staining evaluated the apoptosis and calcification. The level of mitochondria-specific reactive oxygen species (ROS) was quantified with an oxygen radical-sensitive probe-MitoSOX. The potential signaling pathways were investigated by Western blot after the addition of N-acetyl-l-cysteine (NAC). We found that the oxidative stress induced by H2O2increased the apoptosis and subsequently the calcification in the cartilage endplate cells through the ROS/p38/ERK/p65 pathway. The apoptosis and the calcification of the cartilage endplate cells induced by H2O2can be abolished by NAC. These results suggested that regulating the apoptosis and the calcification in the cartilage endplate cells under oxidative stress should be advantageous for the survival of cells and might delay the process of disc degeneration.
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China; Department of Joint and Bone Disease, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai, 200120, China. Electronic address: hycmed@163.com.
Yingchao
Y
Han
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Xinhua
X
Li
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Meijun
M
Yan
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Mingjie
M
Yang
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Shanjin
S
Wang
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Jie
J
Pan
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China. Electronic address: lljunspine@163.com.
Lijun
L
Li
Department of Spinal Surgery, Shanghai East Hospital, Tongji University, School of Medicine, 150 Jimo Road, Shanghai 200120, China.
Jun
J
Tan
D000111:Acetylcysteine / Q000494:pharmacology; D017209:Apoptosis; D002114:Calcinosis / Q000139:chemically induced / Q000378:metabolism / Q000517:prevention & control; D002356:Cartilage / Q000166:cytology / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007403:Intervertebral Disc / Q000166:cytology / Q000378:metabolism; D055959:Intervertebral Disc Degeneration / Q000378:metabolism; D020935:MAP Kinase Signaling System; D016328:NF-kappa B / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*
D016166:Free Radical Scavengers; D016328:NF-kappa B; D016877:Oxidants; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide; D000111:Acetylcysteine
Apoptosis; Calcification; Cartilage endplate (CEP); Intervertebral disc degeneration (IVDD); Oxidative stress; Reactive oxygen species (ROS)