Modulation of H2O2- Induced Neurite Outgrowth Impairment and Apoptosis in PC12 Cells by a 1,2,4-Triazine Derivative

Introduction: Increased oxidative stress is widely accepted to be a factor in the development and progression of Alzheimer’s disease. Triazine derivatives possess a wide range of pharmacological activities including anti-oxidative and anti-in.ammatory actions. In this study, we aimed to investigate the possible protective effect of 3-thioethyl-5,6-dimethoxyphenyl-1,2,4-triazine (TEDMT) on H2O2-induced neurite outgrowth impairment and apoptosis in neuron-like PC12 cells. Methods: We pretreated PC12 cells with 5, 7, and 10 µM of TEDMT followed by adding H2O2 as an oxidative stress agent. Results: We found that TEDMT contributed to up-regulation of Bcl-2, down regulation of Bax protein and reduction of cleaved Caspase-3 and PARP proteins. Moreover, TEDMT could inhibit the phosphorylation of different mitogen activated protein kinases (extracellular signal-regulated kinase, c-Jun N-terminal kinase and P38 mitogen-activated protein kinase). TEDMT induced heat shock protein 70 while decreased heat shock protein 90 level. Besides we measured six different parameters of neurite outgrowth and complexity. We showed that H2O2 increased cell body area, average neurite width and the proportion of bipolar cells, while decreased average neurite length, the numbers of primary neurites and the ratio of the total neurite branching nodes to the total number of primary neurites. Discussion: Interestingly, we found that TEDMT not only protects PC12 cell against H2O2-induced apoptosis, but also defends against the destructive effect of oxidative stress on the criteria of neural differentiation. Protective effect of this compound could represent a promising approach for treatment of neurodegenerative diseases.