The Role Of Chloride Channel In The Hippocampal Neuronal Apoptosis Induced By SIN-1 In Culture

Multiple mechanisms were involved in neuronal apoptosis induced by various factors, including the hypothesis of excitotoxicity, ionic disorder, gene regulation and so on. Nitric Oxide(NO) is an important signaling messenger and has been shown to be involved in many physiological and pathophysiological processes in the brain. Considerable evidence indicates that there is an excessive amount of NO generation in hippocampal neurons following the cerebral hypoxic or ischemic insult, which is responsible for the brain injury. In the previous studies on the ionic mechanisms underlying the neuronal apoptosis in cerebral ischemic damage, attention mainly focused on Ca²⁺, K⁺ and Na⁺channels, less attention was paied on the role of chloride ion. In this study, cultured hippocampal neurons of 12d old(12DIV) from SD rats were used and the cellular models of apoptosis were induced by SIN-1, one kind of NO dornors. In order to analyse the neuronal apoptosis in this experiments, the methods of MTT, a method to analyse the neuronal viability, immunohistochemical fluorescent stain, which is a method to observe the neuronal morphological change, were both used. The methods of Western blot, which is used to detect the activities of caspase-3, the patch clamp in whole-cell configuration, which is used to record the changes of chloride channel currents densities in neuronal apoptosis, were also both used. In current studies, we try to explore the role of chloride channel in the ischemic cerebral injury.