化学低氧诱导剂二氯化钴对PC12细胞自噬的影响  

Effects of cobalt chloride on autophagy in PC12 cells

作　　者：韩苗苗 叶丹蕾 吴玉兰 汪惠丽 HAN Miaomiao;YE Danlei;WU Yulan;WANG Huili(School of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, China)

机构地区：[1]合肥工业大学食品与生物工程学院

出　　处：《合肥工业大学学报（自然科学版）》2019年第10期1411-1414,共4页Journal of Hefei University of Technology：Natural Science

基　　金：国家自然科学基金面上资助项目(81773475)

摘　　要：文章研究化学性低氧诱导剂二氯化钴(CoCl2)对PC12细胞自噬的影响。体外培养PC12细胞,用不同浓度CoCl2处理PC12细胞24 h。首先通过MTT实验选取后续的CoCl2处理浓度,用荧光定量聚合酶链式反应(quantitative polymerase chain reaction, q-PCR)实验检测不同浓度CoCl2处理PC12细胞24 h后细胞内 HIF-1α基因表达水平。用 Western Blot方法检测CoCl2处理后自噬相关蛋白Beclin1、LC3-Ⅱ蛋白表达水平。结果表明,400、800 μmol/L CoCl2处理PC12细胞24 h不仅能显著降低细胞存活率,并且能显著提高 HIF-1α基因表达,即表明用CoCl 2造模确实可以模拟细胞体外缺氧。此外,800 μmol/L CoCl2处理组的Beclin1、LC3-Ⅱ蛋白表达量出现显著降低,表明CoCl 2处理抑制了细胞自噬,并且该种抑制是呈时间依赖的。研究结果表明,CoCl2引起的化学性缺氧会损伤PC12细胞,并且会抑制细胞正常的自噬,该异常的自噬可能参与了CoCl2致PC12细胞的损伤。Effects of cobalt chloride(CoCl2) on autophagy in PC12 cells were studied. PC12 cells were cultured with different concentrations of CoCl2 for 24 h. Firstly, MTT method was used to select the appropriate dosage of CoCl2 for the following investigations. Next, fluorescence quantitative polymerase chain reaction(q-PCR) was used to measure the mRNA level of HIF-1α in CoCl2-exposed PC12 cells. Then, Western Blot was used to investigate the protein levels of autophagy-related protein Beclin1 and LC3-Ⅱ in CoCl 2-exposed PC12 cells. The results showed that the cell viability was significantly reduced in PC12 cells with CoCl2 of 400 μmol/L and 800 μmol/L, and the mRNA level of HIF-1α markedly increased. It demonstrated that the hypoxia model could be established by CoCl2 with the concentrations of 400 μmol/L and 800 μmol/L. Moreover, the protein levels of Beclin1 and LC3-Ⅱsignificantly decreased in CoCl2-exposed PC12 cells with CoCl 2 of 800 μmol/L. It represented that the autophagy was inhibited by CoCl2 exposure and the inhibition was in a time-dependent manner. This study indicated that PC12 cells were impaired and the autophagy was inhibited by CoCl2-induced chemical hypoxia, suggesting that the dysfunctional autophagy may involve in the impairment of PC12 cells induced by CoCl 2 exposure.

关 键 词：二氯化钴 缺氧 PC12细胞 自噬 抑制 

分 类 号：Q56[生物学—生物化学]