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【目的】探讨氯化钴缺氧对大鼠星型胶质细胞TFPI-2基因表达的影响及其可能的调节机制。【方法】将大鼠星型胶质细胞分为对照组、化学缺氧组(加入终浓度为100μmol/L氯化钴培养24 h)。显微镜下观察细胞形态变化,MTT法检测细胞活性,RT-PCR法检测各组缺氧诱导因子-1α(hypoxia induced factor,HIF-1α)、组织因子途径抑制因子-2(tissue factor pathway inhibitor 2,TFPI-2)mRNA表达水平变化,Western blotting检测缺氧前后HIF-1α、TFPI-2、VEGF蛋白表达水平变化。【结果】MTT检测显示,氯化钴缺氧会降低细胞活性,且呈时间依赖性。在显微镜下可观察到细胞缺氧后胞体变小,突触缩短。氯化钴缺氧处理组细胞TFPI-2 mRNA水平显著高于对照组(P<0.05)。Western blotting显示与对照组相比,缺氧后细胞内HIF-1α、TFPI-2、VEGF蛋白表达水平均有显著升高(P<0.05)。【结论】氯化钴缺氧可以激活HIF-1/VEGF缺氧通路,增加SH-SY5Y细胞TFPI-2 mRNA和蛋白表达水平。
【Objective】 To investigate the effect of cobalt chloride depletion on TFPI-2 gene expression in rat astrocytes and its possible regulatory mechanism. [Methods] The astrocytes of rats were divided into control group and chemical hypoxia group (cultured for 24 h with addition of 100 μmol / L cobalt chloride). The morphological changes of the cells were observed under a microscope. The cell viability was measured by MTT assay. The expressions of hypoxia-inducible factor-1α (HIF-1α), tissue factor pathway inhibitor 2 TFPI-2) mRNA expression levels were detected by Western blotting before and after hypoxia HIF-1α, TFPI-2, VEGF protein expression levels. 【Results】 The results of MTT assay showed that hypoxia of cobalt chloride decreased cell activity and was time-dependent. Observed under the microscope cells after hypoxia cells smaller, synapses shortened. The level of TFPI-2 mRNA in cells treated with cobalt chloride hypoxia was significantly higher than that in the control group (P <0.05). Western blotting showed that compared with the control group, the expression of HIF-1α, TFPI-2 and VEGF in hypoxia cells were significantly increased (P <0.05). 【Conclusion】 Cobalt chloride hypoxia can activate HIF-1 / VEGF hypoxic pathway and increase TFPI-2 mRNA and protein expression in SH-SY5Y cells.