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Hydrogen peroxide induces nucleus pulposus cell apoptosis by ATF4/CHOP signaling pathway

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Oxidative stress induces excessive apoptosis resulting in the reduction of intervertebral disc cells, the consequent reduction of extracellular matrix (ECM) synthesis, and compositional changes, which is the pathological basis for intervertebral disc degeneration (IVDD). The present study explored the activating transcription factor 4 (ATF4)/C/EBP homologous protein (CHOP) signaling pathway in the H2O2induced nucleus pulposus (NP) cell apoptosis. Human degenerated intervertebral discs were collected from Lumbar disc surgery. NP cells isolated from the tissues were cultured with H2O2 to induce apoptosis in vitro. Malondialdehyde (MDA) analysis was performed to determine the reactive oxygen species (ROS) of the tissue. Western blot analysis and reverse transcriptionpolymerase chain reaction (RTPCR) were performed to analyze collagen II, ATF4, CHOP, and caspase9 gene expression. Flow cytometry was used to determine the apoptotic ratio of NP cells. siRNA was also used to silence ATF4 and CHOP gene expression. NP tissues in higher degenerated degree underwent much more MDA, expressed less collagen II, more ATF4, CHOP, and caspase9 compared with the mildly degenerated tissues. H2O2 induced NP cell apoptosis by upregulating expression of ATF4, CHOP and caspase9. The silencing of ATF4 or CHOP alleviated NP cell apoptosis by suppressing caspase9 expression. Inhibiting caspase9 did not affect ATF4 and CHOP expression but protected NP cells from apoptosis. In this study, we found H2O2 could promote NP cell apoptosis by activating the ATF4/CHOP signaling pathway resulting in the upregulation of caspase9. Interdict of ATF4, CHOP, or caspase9 contributed to the reduction of apoptosis caused by H2O2.
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Document Type: Research Article

Affiliations: Department of Orthopedics, The First People's Hospital of Fuyang District, Hangzhou, Zhejiang 311400, P.R. China

Publication date: October 1, 2020

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  • Experimental and Therapeutic Medicine aims to ensure the expedient publication, in both print and electronic format, of studies relating to biology, gene therapy, infectious disease, microbiology, molecular cardiology and molecular surgery. The journal welcomes studies pertaining to all aspects of molecular medicine, and studies relating to in vitro or in vivo experimental model systems relevant to the mechanisms of disease are also included.

    All materials submitted to this journal undergo the appropriate review via referees who are experts in this field. All materials submitted follow international guidelines with regard to approval of experiments on humans and animals.
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