小胶质细胞组蛋白去乙酰化酶3在低压低氧诱导的氧化应激中的作用

目的 探究低压低氧对脑内氧化应激的影响及小胶质细胞HDAC3在其中的作用.方法 将野生型小鼠置于模拟海拔高度6000米的低压舱中连续饲养7 d,随后取小鼠海马组织并提取RNA,实时定量聚合酶链反应(real-time polymerase chain reaction,real-time PCR)检测氧化应激标志物诱导...

Role of microglial histone deacetylases 3 in hypobaric hypoxia-induced oxidative stress

Objective To investigate the effect of hypobaric hypoxiaon the brain tissue oxidative stress level and whether microglial HDAC3 is involved in this process. Methods First, wild type mice were treated in a hypobaric chamber at a simulated altitude of 6 000 meters for 7 days.The mRNA levels of hypoxia inducible factors-1α (HIF-1α), inducible nitric oxide synthase(iNOS), superoxidase dismutase(SOD) and histone deacetylases 3 (HDAC3) in the hippocampus were analyzed by real-time quantitative polymerase chain reaction (real-time PCR). Then, BV2 cells were treated in atmosphere with 0.1% O₂ in a hypoxia incubator and the mRNA and protein levels of iNOS, SOD, HDAC3 and HIF-1α were determined by Western blotting and real-time PCR.The levels of reactive oxygen species (ROS) were analyzed by flow cytometry.Furthermore, HDAC3-silenced and wild type BV2 cells were cultured in 0.1% O₂ atmosphere and the levels of ROS, iNOS and SOD were analyzed as above. Last, microglial HDAC3 conditional knockout mice and wild type mice were treated in hypoxia atmosphere, and the mRNA level of iNOS were analyzed by real-time PCR. Results The mRNA levels of iNOS, SOD and HDAC3 were upregulated in the hippocampus from mice underwent hypobaric hypoxia.The in vitro experiments indicated that the protein levels of iNOS, HDAC3 and HIF-1α were increased in BV2 cells treated 0.1% O₂, and the mRNA levels of iNOS and SOD were increased as well. By flow cytometry, we found the level of ROS was increased when BV2 cells were cultured in hypoxia atmosphere. Inhibition of HDAC3 significantly reduced the production of hypoxia-induced ROS, as well as decreased expression of SOD and iNOS. In addition, knockout of HDAC3 in microglia inhibited the hypoxia-induced upregulation iNOS. Conclusion The hypoxia treatment resulted in increased oxidative stress in the hippocampus and microglia, and inhibition of microglial HDAC3 ameliorated hypoxia-induced oxidative stress.