Impacts of AMPK activators and metal stressors on energy levels of Neuro 2a cells

Abstract

ATP is the most critical molecule for the maintenance and survival of cells in general, especially for the brain and neuronal cells to support essential processes including ion homeostasis, synaptic functions, neurotransmitter production and release. A considerable body of clinical evidence has shown that energy deficit is a very early hallmark of the Alzheimer's disease (AD) brain occurring even prior to clinical symptoms and consistently in association with the progression of the disease. Viewed from this perspective, AMP-activated protein kinase (AMPK), a key enzyme maintaining energy balance in the whole-body level and single-cell level, was hypothesized as a potential target for AD therapeutics. Besides that, another angle of energy disturbance focusing on risk factors of AD - metal stressors was also investigated in the thesis. The project aimed to provide solid scientific evidence for the protective effects of AMPK activators and potential risks of metal stressors in the energetic aspect in current neuronal culture models. A large portion of the project related to optimization of an HPLC method which provided an effective tool to determine numerous nucleotides simultaneously. Among a couple of AMPK activators tested in the project, Resveratrol manifested exciting pro-energetic benefits in the neuronal culture cells rapidly and lastingly. The results show that Resveratrol induced a strongly significant increase in the levels of the most important energy carriers, ATP and GTP, in Neuro 2a cells in a dose-dependent manner, even from the early time-point of 1 hour and extended up to 48 hours. In the experiments on metal stressors, it was discovered that whilst zinc exhibited its cytotoxicity in an adenosine and guanosine nucleotideindependent mechanism, iron revealed the potential harm on the energy generation. Since iron accumulation in the brain has been shown to be a common feature of neurodegenerative diseases including AD and Parkinson's disease (PD) in the literature, excessive iron exposure in AD and PD brains might contribute to the disturbance of energy production to be more severe. Collectively, we promote that Resveratrol and strategies against iron overload in the brain should be considered as potential candidates for prevention and treatment of energy depletion in AD.

Date of Award	2012
Original language	English