LPS诱导巨噬细胞凋亡的研究

Study on the mechanism of LPS-induced apoptosis in macrophage

目的 探讨脂多糖 (LPS)诱导巨噬细胞发生凋亡的分子机理及信号传导通路。方法 采用夹心ELISA法检测巨噬细胞NF κB的核内浓度 ;硝酸还原酶法测定细胞培养液中NO含量 ;吖啶橙荧光染色及凋亡电泳试验显示巨噬细胞凋亡情况。结果 LPS刺激可诱导巨噬细胞的NF κB活化、NO分泌并最终导致细胞凋亡 ,而特异性PKC抑制剂 (CalC)和NF κB阻断剂 (PDTC)能在不同程度上抑制LPS的生物活性作用。结论 LPS诱导肺巨噬细胞凋亡的信号传导通路有PKC、NF κB的参与 ,而NO等生物活性介质在其中发挥了重要作用。

Objective To study the molecular mechanism and signal transduction pathway of LPS-induced macrophage apoptosis. Methods NF-κB level in nuclear protein extraction of AMs (alveolar macrophage) was detected with sandwich ELISA, NO content in cell culture medium was quantified with nitric acid reductase assay. Morphologic change of AMs in apoptosis observed with acridine orange (A.O) staining and apoptotic fragmentation at genome DNA of AMs detected with electrophoresis assay. Results LPS-induced NF-κB activation and NO production of AMs which cause apoptosis were decreased and inhibited by the pretreatment of Cal C (calphostin C) and PDTC (pyrrolidine dithiocarbamate). Conclusion PKC (protein kinase C), NF-κB and NO are involved in the signal transduction pathway of LPS-induced macrophage apoptosis.