脯氨酰羟化酶抑制剂对PC12细胞 SDF-1α表达的影响

目的 研究脯氨酰羟化酶抑制剂对基质细胞衍生因子-1α(stromal cell-derived factor-1 alpha, SDF-1α)表达的影响及可能的机制。方法 体外培养PC12细胞,予以不同浓度氯化钴(CoCl2)、去铁胺(DFO)、二甲基乙二酰基甘氨酸(DMOG)处理。CCK-8法检测细胞活性,RT-PCR检测药物诱导后细胞内SDF-1α mRNA的表达,Western印迹法和ELISA法分别检测低氧诱导因子-1α(hypoxia inducible factor 1α, HIF-1α)、SDF-1α蛋白表达。结果 CCK-8法结果显示,低浓度CoCl2能够促进PC12细胞的增殖能力,但随着浓度增高,DFO、CoCl2、DMOG均能使细胞增殖能力降低。RT-PCR结果显示,DFO、CoCl2、DMOG各组细胞内SDF-1α mRNA表达减少。Western印迹法结果显示,CoCl2、DFO和DMOG组HIF-1α表达量较对照组显著增高。ELISA法结果显示,CoCl2、DFO、DMOG组SDF-1α水平均显著下降。结论 脯氨酰羟化酶抑制剂CoCl2、DFO和DMOG均可上调HIF-1α的表达,下调SDF-1α的表达。
Objective To investigate the effects and underlying mechanisms of prolyl hydroxylase domain (PHD) inhibitors on the expressions of SDF-1α in PC12 cells. Methods Cultured rat pheochromocytoma PC12 cells were treated with different concentrations of PHD inhibitors cobalt chloride(CoCl2),  desferrioxamine (DFO) or dimethyloxalylglycine (DMOG), respectively. Cell viability was determined with cell counting kit-8 (CCK-8); the expression of SDF-1α mRNA was detected with quantitative real time PCR; the expression of HIF-1α and SDF-1α protein was measured with Western blotting and ELISA assays, respectively. Results CCK-8 showed that low concentration of CoCl2 promoted cellular growth; however, DFO, DMOG and higher concentration of CoCl2 inhibited cell growth with concentration increasing. The mRNA level of SDF-1α was down regulated in a concentration dependent manner by DFO, CoCl2 and DMOG. Western blotting showed that HIF-1α levels in PC12 cells treated with CoCl2, DFO or DMOG were higher than that in control group. ELISA analysis showed that SDF-1α expression was significantly lower in PC12 cells treated with DFO, CoCl2 or DMOG. Conclusion Prolyl hydroxylase domain (PHD) inhibitors up-regulate the level of HIF-1α and down-regulate the level of SDF-1α in rat pheochromocytoma PC12 cells