大鼠肺巨噬细胞iNOS电镜细胞化学定位显示法

The demonstration of NOS in rat alveolar macrophage with electron microscopic NADPH-d cytochemistry

目的 对大鼠肺泡巨噬细胞NOS进行细胞内定位显示。方法 由大鼠支气管肺泡灌洗液中获取肺泡巨噬细胞 ,富集制备成单层细胞爬片。在 37℃ ,5%CO2 的二氧化碳培养箱中 ,用LPS ( 1g L)刺激 2 4h以诱导肺泡巨噬细胞NOS表达。采用NADPH d电镜细胞化学术对肺泡巨噬细胞NOS进行定位显示。结果 活化的AM胞浆内可见大量颗粒状电子密度沉淀。结论 NADPH

Objectives:This study was designed to localize the nitric oxide synthase (NOS) in rat alveolar macrophages (AM) in vitro. Methods:Special techniques of bronchoalveolar lavage were used to harvest wistar rat AM. After cells enrichment, AM on glass cover slips were cultured 24h with LPS (1g/L) or saline (control group) at 37℃ in 5% CO 2 incubator. The AM monolayers were then stained with electron microscopic NADPH diaphorase (NADPH d) cytochemistry. Results:The activated AM demonstrated that electron dense grains were throughout cytoplasmic area. Conclusions:With NADPH d cytochemistry, the AM inducible NOS is a kind of cytoplasmic enzyme.