目的:观察胞浆内线粒体 DNA(mitochondrial DNA,mtDNA)在脂多糖(lipopolysaccharide,LPS)诱导的NR8383肺泡巨噬细胞炎症反应时的动态变化,以初步探讨胞浆内 mtDNA在肺泡巨噬细胞炎症反应中的作用。方法将体外培养的大鼠肺泡巨噬细胞(NR8383)用终浓度为1μg·mL-1的 LPS刺激0、3、6、12和24 h后收集细胞培养上清液及细胞,采用差速离心法分离胞浆中的mtDNA,实时荧光定量聚合酶链反应(real-time PCR)检测胞浆中mtDNA表达情况,采用酶联免疫吸附法(ELISA)检测细胞培养上清液中 IL-1β蛋白水平。结果 LPS刺激后3、6、12和24 h IL-1β及胞浆中mtDNA的含量均显著高于0 h组(P<0.01)。结论 LPS刺激肺泡巨噬细胞后,胞浆中mtDNA含量升高,推测其可能参与了肺泡巨噬细胞的炎症反应。
Objective To observe the dynamic changes in cytoplasmic mitochondrial DNA (mtDNA)in lipopolysaccharide (LPS)-induced alveolar macrophage inflammatory response,and to explore the role of mtDNA in alveolar macrophage inflammatory response.Methods Alveolar macrophages(NR8383)were stimulated with LPS(1μg·mL-1)in vitro.Culture supernatants and cells were collected after treatment for 0,3,6,12 and 24 hours,and cytoplasmic mtDNA was isolated by differential centrifugation.The expression of mtDNA was detected by real-time PCR, and the level of interleukin-1β(IL-1β)in the supernatants was measured by ELISA.Results The levels of IL-1βin supernatants and mtDNA in cytoplasm after LPS stimulation for 3,6,12 and 24 hours were significantly higher than those at the beginning of LPS stimulation (P<0.01).Con-clusion The expression of mtDNA in the cytoplasm of alveolar macrophages increases after stim-ulation with LPS,suggesting that mtDNA may be involved in alveolar macrophage inflammatory response.