Study On The Effect Of On TNFAIP1 H₂O₂-Induced Neuronal Injury

In recent years,neurodegenerative diseases incidence has risen dramatically,especially in an ageing population.Due to the complexity of its pathogenesis,and the pathophysiology of the exact mechanism is not yet clear,so there is no ideal drug prevention and cure occureed.Which is caused by a variety of reasons,and puts forward the multiple hypothesis,The representative hypothesis including neurotransmitter imbalance theory,theory of oxidative stress,mitochondrial dysfunction theory,theory of excitatory neurotoxicity,abnormal protein accumulation theory,gene mutation,abnormal inflammatory process,immune and cell apoptosis theory,etc.In recent years,more and more data demonstrate that the most represent hypothesis is that mitochondrial dysfunction and oxidative stress interact with other several mechanisms,and oxidative stress and mitochondria might be the main triggering factor.When intracellular reactive oxygen species（reactive oxygen species,ROS） accumulate,oxidation level exceeds the antioxidant effect,oxidative stress will occure.ROS plays an important role in cell signal transduction,involved in many physiological and pathological process.And hydrogen peroxide（H₂O₂） is the product of metabolism in the body,is also a kind of ROS,plays a key role in inducing neurons injury and even death.H₂O₂ has been applied to many studies of the oxidative damage,and H₂O₂induced cell oxidative damage has become one of the important tools of exploring neuronal injury.TNFAIP1 is the protein induced by tumor necrosis factor alpha and interleukin 6.TNFAIP1 may play an important role in DNA repair,DNA replication,signaling transduction and in the process of many diseases.Early research has shown that TNFAIP1 mediated Aβinducing nerve injury.In order to further explore the role of TNFAIP1 induced neurotoxicity,we use N2a neuroblastoma cells in mice with H₂O₂ treament,simulating the model of oxidative stress,to explore TNFAIP1 in H₂O₂ induced the process of nerve injury.Using western blot experiments,we found that H₂O₂ could rise TNFAIP1 in dose dependent.Through the determined by MTT experiment,we discovered TNFAIP1 mediated the N2a cells viability decline induced by H₂O₂.In addition,by reactive Oxygen Species detection Kit（Ractive Oxygen Species Assay Kit） and Mitochondrial membrane potential detection Kit（Mitochondrial be potential Assay Kit with JC-1） respectively to detect ROS（Ractine Oxygen Species） and Mitochondrial membrane potential,find TNFAIP1 mediated hydrogen peroxide promoted the production of ROS and decreased Mitochondrial membrane potential.On the basis of these results,we also preliminary studied the signal pathway,found TNFAIP1 cut the Bcl-2 and the phosphorylation of CREB,and raised the Cleaved-Caspase3.Therefore,we suspect that TNFAIP1 may inhibit the CREB signaling pathway,resulting in a decline in mitochondrial membrane potential and promoting H₂O₂-induced neuronal injury.