Wang Changliang, Wang Xiaolong, Li Yan, Xia Zhixiu, Liu Yang, Yu Hao, Xu Guohui, Wu Xu, Zhao Rui, Zhang Guohua
Department of Forensic Pathology, School of Forensic Medicine.
The People's Procuratorate of Liaoning Province Judicial Authentication Center.
Neuroreport. 2019 Apr 10;30(6):397-403. doi: 10.1097/WNR.0000000000001214.
Chronic ethanol (EtOH) exposure can cause intracellular Ca overload by stimulating calcium channel receptors and trigger apoptosis of neurons. NCX3 may play a cytoprotective role in intracellular Ca excretion. In this study, the effect of EtOH on NCX3 was analyzed by observing NCX3 expression in the hippocampus of chronic EtOH-exposed male C57BL/6 mice. Mice were divided into a control group, a 10% EtOH group, and a 20% EtOH group for 30, 60, and 90 days. Behavioral changes were observed using the Morris water maze. The protein and mRNA expressions of NCX3 and their distribution in the hippocampus were observed by western blotting, quantitative PCR, and immunohistochemistry staining. The results showed that EtOH exposure exerted a significant adverse effect on the spatial memory capacity of mice. Increased expression of calpain-1 and cleaved caspase-3 proteins indicated increased apoptosis. The expression of NCX3 in the hippocampus was downregulated after exposure to EtOH (except 10% EtOH for 30 days) and this inhibition was time and dose dependent with EtOH exposure. The level of p-Akt, which is an upstream regulation factor of NCX3, showed a trend similar to that of NCX3 protein. Chronic EtOH exposure reduced the expression of NCX3 in the hippocampus of male C57BL/6 mice, increasing intracellular calcium and apoptosis, resulting in spatial memory impairment in mice.
慢性乙醇（EtOH）暴露可通过刺激钙通道受体导致细胞内钙超载，并引发神经元凋亡。NCX3可能在细胞内钙排泄中发挥细胞保护作用。在本研究中，通过观察慢性乙醇暴露的雄性C57BL/6小鼠海马中NCX3的表达，分析了乙醇对NCX3的影响。将小鼠分为对照组、10%乙醇组和20%乙醇组，分别处理30、60和90天。使用Morris水迷宫观察行为变化。通过蛋白质印迹法、定量PCR和免疫组织化学染色观察NCX3的蛋白质和mRNA表达及其在海马中的分布。结果表明，乙醇暴露对小鼠的空间记忆能力产生了显著的不利影响。钙蛋白酶-1和裂解的半胱天冬酶-3蛋白表达增加表明细胞凋亡增加。乙醇暴露后海马中NCX3的表达下调（10%乙醇处理30天除外），且这种抑制与乙醇暴露的时间和剂量相关。作为NCX3上游调节因子的p-Akt水平呈现出与NCX3蛋白相似的趋势。慢性乙醇暴露降低了雄性C57BL/6小鼠海马中NCX3的表达，增加了细胞内钙和细胞凋亡，导致小鼠空间记忆受损。
