Synergistic Utilization of Necrostatin-1 and Z-VAD-FMK Efficiently Promotes the Survival of Compression-Induced Nucleus Pulposus Cells via Alleviating Mitochondrial Dysfunction.

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  • Additional Information
    • Source:
      Publisher: Wiley Country of Publication: United States NLM ID: 101600173 Publication Model: eCollection Cited Medium: Internet ISSN: 2314-6141 (Electronic) NLM ISO Abbreviation: Biomed Res Int Subsets: MEDLINE
    • Publication Information:
      Publication: 2024- : [Hoboken, NJ] : Wiley
      Original Publication: New York, NY : Hindawi Pub. Co.
    • Subject Terms:
      Amino Acid Chloromethyl Ketones/*pharmacology ; Imidazoles/*pharmacology ; Indoles/*pharmacology ; Membrane Potential, Mitochondrial/*drug effects ; Nucleus Pulposus/*drug effects ; Oxidative Stress/*drug effects; Animals ; Apoptosis ; Apoptosis Regulatory Proteins/metabolism ; Autophagy ; Cadaverine/analogs & derivatives ; Cadaverine/pharmacology ; Cell Death ; Cell Survival ; Compressive Strength ; L-Lactate Dehydrogenase/metabolism ; Mitochondria/metabolism ; Nucleus Pulposus/cytology ; Pressure ; Propidium/chemistry ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species
    • Abstract:
      We recently reported that necroptosis contributed to compression-induced nucleus pulposus (NP) cells death. In the current study, we investigated the regulative effect of necroptosis inhibitor Necrostatin-1 on NP cells apoptosis and autophagy. Necrostatin-1, autophagy inhibitor 3-Methyladenine and apoptosis inhibitor Z-VAD-FMK were employed, and NP cells were exposed to 1.0 MPa compression for 0, 24 and 36 h. Necroptosis-associated molecules were measured by Western blot and RT-PCR. Autophagy and apoptosis levels were evaluated by Western blot and quantified by flow cytometry after monodansylcadaverine and Annexin V-FITC/propidium iodide staining, respectively. The cell viability and cell death were also examined. Furthermore, we measured mitochondrial membrane potential (MMP), mitochondrial permeability transition pore (MPTP) and indices of oxidative stress to assess mitochondrial dysfunction. The results established that Necrostatin-1 blocked NP cells autophagy, and 3-Methyladenine had little influence on NP cells necroptosis. The Necrostatin-1+3-Methyladenine treatment exerted almost the same role as Necrostatin-1 in reducing NP cells death. Necrostatin-1 restrained NP cells apoptosis, while Z-VAD-FMK enhanced NP cells necroptosis. The Necrostatin-1+Z-VAD-FMK treatment provided more prominent role in blocking NP cells death compared with Necrostatin-1, consistent with increased MMP, reduced opening of MPTP and oxidative stress. In summary, the synergistic utilization of Necrostatin-1 and Z-VAD-FMK is a very worthwhile solution in preventing compression-mediated NP cells death, which might be largely attributed to restored mitochondrial function.
      Competing Interests: The authors declare that they have no conflict of interest.
      (Copyright © 2020 Songfeng Chen et al.)
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    • Accession Number:
      0 (Amino Acid Chloromethyl Ketones)
      0 (Apoptosis Regulatory Proteins)
      0 (Imidazoles)
      0 (Indoles)
      0 (Reactive Oxygen Species)
      0 (benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone)
      0 (necrostatin-1)
      36015-30-2 (Propidium)
      EC 1.1.1.27 (L-Lactate Dehydrogenase)
      I9N81SC5HD (monodansylcadaverine)
      L90BEN6OLL (Cadaverine)
    • Publication Date:
      Date Created: 20201230 Date Completed: 20210608 Latest Revision: 20210608
    • Publication Date:
      20250114
    • Accession Number:
      PMC7738793
    • Accession Number:
      10.1155/2020/6976317
    • Accession Number:
      33376733