高氧抑制LPS/ATP诱导的骨髓源性巨噬细胞的焦亡

Hyperoxia Inhibits LPS/ATP-Induced Pyroptosis of Bone Marrow Derived Macrophages

目的:研究不同浓度的氧气在LPS/ATP诱导的骨髓源性巨噬细胞焦亡中的作用。方法:提取C57BL/6小鼠的骨髓源性巨噬细胞,用1μg/ml脂多糖(LPS)刺激细胞24 h,用5 mM三磷酸腺苷(ATP)刺激细胞4 h,酶联免疫吸附测定法(ELISA)检测细胞培养上清液中IL-1β水平的变化。用5 mM ATP刺激细胞后,给予细胞40%、60%和100%的氧气处理1.5 h,ELISA检测细胞培养上清液中IL-1β水平的变化。结果:1μg/mL LPS和5 mM ATP先后刺激下,骨髓源性巨噬细胞培养上清液中IL-1β的水平明显升高(P<0.001),用caspase-1特异性抑制剂AC-YVAD-CMK刺激骨髓源性巨噬细胞后IL-1β水平明显降低(P<0.001)。5 mM ATP刺激之后给予细胞不同浓度的氧气干预1.5 h后,细胞培养上清液中IL-1β的水平明显下降。结论:高氧抑制LPS/ATP诱导的骨髓源性巨噬细胞的焦亡。

Objective： To explore the effect of different concentrations of oxygen in LPS/ATP induced pyroptosis of bone marrow derived macrophages. Methods： To extract the bone marrow derived macrophages of C57BL/6 mice and cells were primed with 1 ~g/ml LPS for 24 h, and then treated with 5 mM ATP for 4 h. IL-1β levels in the cell supematants were determined using a commercially available mouse IL-1β ELISA kit. Give cells 40 %, 60 % or 100 % oxygen inhalation after 5 mM ATP stimulation and determine the IL-1β levels in the cell supernatants. Results： ELISA test showed IL-1β 1 evels in the bone marrow derived macrophages supematants were greatly increased after the stimulation of 1μg/ml LPS and 5 mM ATP （P〈0.001） and were declined after the stimulation of caspase-1 inhibitor AC-YVAD-CMK （P〈0.001）. ELISA test also showed IL-1β levels in the bone marrow derived macrophages supernatants were notably declined after the 1.5 hours stimulation of 40 %, 60 % and 100 % oxygen inhalation followed the stimulation of 5 mM ATP. Conclusions： Hyperoxia inhibits the pyroptosis of bone marrow derived macrophages.