Apocynin Attenuates Cobalt Chloride-Induced Pheochromocytoma Cell Apoptosis by Inhibiting P38-MAPK/Caspase-3 Pathway

Cell Physiol Biochem. 2018;48(1):208-214. doi: 10.1159/000491720. Epub 2018 Jul 13.

Abstract

Background/aims: Apocynin, a nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor, has been identified as a potential neuroprotectant. In this study, we aimed to investigate the protective effect of apocynin against cobalt chloride (CoCl2)-induced pheochromocytoma (PC12) cell apoptosis.

Methods: The PC12 cell culture was pretreated with apocynin and/or SB203580 (p38 mitogen-activated protein kinase [p38-MAPK] inhibitor) at different time points prior to CoCl2 incubation. The cell viability, apoptosis rate, DAN damage, and antioxidant activity were detected using cell counting kit-8 (CCK-8), flow cytometry, enzyme-linked immunosorbent assay (ELISA), and comet assay respectively. The protein and mRNA expressions of p38-MAPK and caspase-3 in the cells were measured by qRT-PCR and Western blotting.

Results: Apocynin inhibited CoCl2-mediated apoptosis, reduced oxidative stress, and down-regulated the expression of p38-MAPK and caspase-3.

Conclusions: Our findings show that apocynin attenuated CoCl2-induced apoptosis by potently restraining p38-MAPK-caspase-3 signaling pathway in PC12 cells, suggesting that apocynin may be a potent prophylactic reagent against CoCl2-mediated PC12 cell apoptosis.

Keywords: Apocynin; Apoptosis; Caspase-3; Cobalt chloride; NADPH oxidase; Pheochromocytoma cell; p38-MAPK.

MeSH terms

  • Acetophenones / pharmacology*
  • Adrenal Gland Neoplasms / metabolism
  • Adrenal Gland Neoplasms / pathology
  • Animals
  • Apoptosis / drug effects*
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cell Survival / drug effects
  • Cobalt / pharmacology*
  • DNA Damage / drug effects
  • Gene Expression / drug effects
  • Imidazoles / pharmacology
  • Malondialdehyde / metabolism
  • PC12 Cells
  • Pheochromocytoma / metabolism
  • Pheochromocytoma / pathology
  • Pyridines / pharmacology
  • Rats
  • Signal Transduction / drug effects*
  • Superoxide Dismutase / metabolism
  • p38 Mitogen-Activated Protein Kinases / genetics
  • p38 Mitogen-Activated Protein Kinases / metabolism

Substances

  • Acetophenones
  • Imidazoles
  • Pyridines
  • Cobalt
  • Malondialdehyde
  • acetovanillone
  • Superoxide Dismutase
  • p38 Mitogen-Activated Protein Kinases
  • Caspase 3
  • cobaltous chloride
  • SB 203580