低氧预适应对HT22细胞自噬的影响

Effects of hypoxic preconditioning on autophagy of HT22 cells

目的:研究低氧预适应(HPC)对小鼠海马HT22细胞自噬的影响。方法:将HT22细胞并分为对照组(control)、低氧组(hypoxia)、低氧预适应组(HPC)和低氧预适应与3-甲基腺嘌呤联合处理组(HPC+3-MA)。MTS法检测HT22细胞活性,采用real time RT-PCR检测HT22细胞中自噬及微管相关蛋白1轻链3(LC3)mRNA表达,利用Western Blot检测LC3蛋白的表达及LC3Ⅱ/LC3Ⅰ的比值变化,转染GFP-LC3进一步检测LC3Ⅱ表达。结果:HPC可增加低氧状态下HT22细胞的活性,与HPC组相比,HPC+3-MA组HT22细胞活性降低;HPC可以诱导低氧状态下HT22细胞中LC3的mRNA的表达,并增加LC3Ⅱ/LC3Ⅰ的比值。结论:HPC通过促进低氧状态下HT22细胞自噬提高细胞活性。

Objective:The aim of this study was to explore the effect of hypoxic preconditioning(HPC)on autophagy in mouse hippocampal neuronal HT22 cells.Methods:HT22 cells were divided into four groups:control group,hypoxia group,hypoxia preconditioning group(HPC group),and hypoxic preconditioning which treated with 3-Methyladenine group(HPC+3-MA).Cell viability was measured by MTS method.Microtubule-associated protein1 light chain 3(LC3)mRNA level was detected by real time RT-PCR.The protein expression level of LC3 and LC3Ⅱ/LC3Ⅰratio were tested by Western Blot.The LC3Ⅱwas observed in cells which transfected with GFP-LC3.Results:HPC can increase the viability of HT22 cells under hypoxia.Compared with that in the HPC group,the viability of HT22 cells in the HPC+3-MA group was reduced.HPC can induce the expression of LC3 mRNA in HT22 cells under hypoxia and increased LC3Ⅱ/LC3Ⅰratio.Conclusion:HPC can improve cell viability by promoting autophagy in HT22 cells under hypoxia.