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

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

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-inflammatory 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. We pretreated PC12 cells with 5, 7, and 10 µM of TEDMT followed by adding H2O2 as an oxidative stress agent. 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. 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

Antibody Therapy Targeting RAN Proteins Rescues C9 ALS/FTD Phenotypes in C9orf72 Mouse Model

The intronic C9orf72 G4C2 expansion, the most common genetic cause of ALS and FTD, produces sense- and antisense-expansion RNAs and six dipeptide repeat-associated, non-ATG (RAN) proteins, but their roles in disease are unclear. We generated high-affinity human antibodies targeting GA or GP RAN proteins. These antibodies cross the blood-brain barrier and co-localize with intracellular RAN aggregates in C9-ALS/FTD BAC mice. In cells, α-GA1 interacts with TRIM21, and α-GA1 treatment reduced GA levels, increased GA turnover, and decreased RAN toxicity and co-aggregation of proteasome and autophagy proteins to GA aggregates. In C9-BAC mice, α-GA1 reduced GA as well as GP and GR proteins, improved behavioral deficits, decreased neuroinflammation and neurodegeneration, and increased survival. Glycosylation of the Fc region of α-GA1 is important for cell entry and efficacy. These data demonstrate that RAN proteins drive C9-ALS/FTD in C9-BAC transgenic mice and establish a novel therapeutic approach for C9orf72 ALS/FTD and other RAN-protein diseases