淫羊藿甙对小鼠腹腔巨噬细胞体外增殖、吞噬和产生NO的影响

无菌分离小鼠腹腔巨噬细胞,制备单细胞悬液,加入不同终浓度的淫羊藿甙孵育4 h后,加入细菌脂多糖LPS(终浓度20 mg/L)48 h后以MTT法检测其增殖;加药孵育4 h后,分别加入直径1μm和2μm的微球(终浓度1×1010/L),5 h后利用流式细胞术(FCM)检测吞噬情况;加入LPS(终浓度20 mg/L)24 h后Griess试剂盒检测巨噬细胞NO的量。结果显示ICA在终浓度1.5、3.0μmol/L时能明显抑制LPS刺激的巨噬细胞增殖(P<0.01)和NO的产生,并促进其吞噬微球的功能。提示ICA可能抑制LPS信号转导从而抑制巨噬细胞增殖,并且通过抑制其活化,下调iNOS有关的炎症因子,使NO减少;对于未活化的巨噬细胞,ICA可能通过上调其吞噬功能,增强固有免疫系统来抵御病原体侵入。

Effect of icariin on proliferation,phagocytosis and NO production of murine peritoneal macrophages in vitro

The single cell suspension of murine peritoneal macrophages was prepared under sterile condition and stimulated with LPS at a final concentration of 20 mg/L.Then,the proliferation of macrophages was measured by MTT assay 48 hours later,Macrophages in this cell suspension were treated with different concentrations of icariin(ICA) after 4 hours incubation and addition of microbeads with a diameter of 1 to 2 μm(final concentration 1×1010/L),and the condition of phagocytosis by macrophages was observed by flow cytometry(FCM) 5 hours later.The amount of NO produced in macrophages 24 hours later was detected by Griess kit.It was found that both 1.5 and 3.0 μmol/L of ICA inhibited proliferation and NO production of macrophages stimulated by LPS and promoted the ability of macrophages to engulf microbeads.These results suggest that ICA can inhibit the transduction of LPS-signals leading to proliferation of macrophages and also inhibit its activation by down-regulating the iNOS-involved inflammatory factors,thus to decrease the production of NO.However,to the un-activated macrophages,ICA up-regulates phagocytic functions and promotes the innate immunity to resist invasion of pathogens.