缺氧缺糖/复氧复糖对大鼠皮质及海马神经元ClC-3氯离子通道表达的影响

The expression of ClC-3 chloride channel after hypoxia/hypoglycemia and reoxygenation in rat cortical and hippocampal neurons

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。

Objective ： To investigate the expression of voltage-gated chloride channel 3 （ C1C-3 ） mRNA and protein in the cortical and hippocampal neurons of rat after hypoxia/hypoglycemia and reoxygenation. Methods ： The original gener- ation of cortical and hippocampal neurons were cultured for 8 days and then divided into control group, hypoxia/hypogly- cemia and reoxygenation group. Cells in hypoxia/hypoglycemia and reoxygenation group were treated with reoxygenation after deprivation of oxygen and glucose for 30 rain except control group. The expression levels of C1C-3 mRNA and protein were measured at 6, 24, 48, 72 h after reoxygenation by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively. Results： C1C-3 mRNA and protein were low expressed in rat original generation cultured cor- tical and hippocampal neurons. Compared with control group, C1C-3 mRNA and protein expression after hypoxia/hypogly- cemia and reoxygenation in cortical neurons increased at 24 h and achieved highest level at 48 h, then reduced after 72 h （ P 〈 0.05 ）. C1C-3 mRNA expression in original generation cultured hippocampal neurons began to increase at 6 h and reached the peak value at 24 h to 48 h （P 〈 0.05 ） , then it decreased gradually, but was still higher than that of the con- trol group at 72 h （ P 〈 0.05 ）. C1C-3 protein expression in hippocampal neurons increased slightly at 24 h and was still highly expressed at 72 h （ P 〈 0.05 ）. Conclusion ： The upregulation of C1 C-3 expression in rat cortical and hippocampal neurons may enhance the viability of cells against oxidative stress and inflammation, but promote the possibility of apoptosis.