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目的:研究原代培养大鼠前额叶皮质和海马神经元缺氧缺糖/复氧复糖后电压门控氯离子通道3(voltage-gated chloride channel 3,ClC-3)在mRNA和蛋白水平的表达。方法:取原代培养8 d的大鼠皮质和海马神经元,随机分为对照组和模型组。模型组缺氧缺糖30 min后再复氧复糖,于复氧复糖后6、24、48、72 h采用反转录酶-聚合酶链锁反应(RT-PCR)、Western blot技术检测ClC-3 mRNA及蛋白水平的表达。结果:原代培养大鼠皮质和海马神经元ClC-3 mRNA及蛋白水平呈低水平表达。缺氧缺糖/复氧复糖24 h后培养皮质神经元ClC-3mRNA及蛋白水平开始上调,48 h仍然高表达,72 h后下降(P<0.05)。海马神经元ClC-3 mRNA在缺氧缺糖/复氧复糖6 h后即开始升高,高峰持续从24～48 h(P<0.05),72 h下降至略高于正常水平(P<0.05)。海马神经元ClC-3蛋白在24 h后表达开始逐渐升高,至72 h仍高表达(P<0.05)。结论:C1C-3通道表达上调可能增强复氧复糖后细胞对氧化应激、炎性反应的同时也促进细胞凋亡。
AIM: To investigate the mRNA and protein levels of voltage-gated chloride channel 3 (ClC-3) after primary culture of rat prefrontal cortex and hippocampal neurons under hypoxia / hypoglycemia / reoxygenation expression. Methods: Primary cultured cortical and hippocampal neurons of 8-day-old rats were randomly divided into control group and model group. After reoxygenation for 30 min in hypoxia-deprivation glucose model group, RT-PCR at 6,24,48 and 72 h after reoxygenation and reoxygenation were detected by Western blot ClC-3 mRNA and protein expression. Results: The mRNA and protein levels of ClC-3 in primary cultured cortical and hippocampal neurons were low. The mRNA and protein levels of ClC-3 in cortical neurons began to increase after 24 hours of hypoxia / hypoglycemia / reoxygenation, and remained high at 48 hours and decreased at 72 hours (P <0.05). The ClC-3 mRNA of hippocampal neurons began to rise after 6 hours of hypoxia / hypoglycemia / reoxygenation, the peak continued from 24 to 48 hours (P <0.05), and decreased to slightly higher than normal level (P < 0.05). The expression of ClC-3 protein in hippocampal neurons began to increase gradually after 24 h and remained high at 72 h (P <0.05). CONCLUSION: Up-regulation of C1C-3 channel may enhance the cellular responses to oxidative stress and inflammatory reaction as well as promote cell apoptosis.