目的探讨枸橼酸铁铵(FAC)对脂多糖(LPS)激活的大鼠小胶质细胞乳铁蛋白(Lf)表达影响。方法分别使用FAC、LPS、FAc+LPS处理原代培养的大鼠小胶质细胞,应用酶联免疫吸附试验方法检测细胞上清液中Lf的含量;采用实时荧光定量PCR方法检测细胞LfmRNA的表达。结果与对照组相比较,LPS处理组U蛋白和mRNA表达水平均明显增高,差异有显著性(F=458.153、78.884,q=18.786、10.683,P〈O.05);FAC+LPS组Lf蛋白表达水平较LPS组明显增高,差异有显著性(q=12.831,P〈O.05),但LfmRNA水平未见明显升高;FAC处理组与对照组相比较,Lf蛋白和mRNA表达水平差异无显著性(P〉O.05)。结论单独FAC不能引起小胶质细胞Lf的表达增高;LPS激活的小胶质细胞合成和释放Lf增多;高铁状态提高激活的小胶质细胞Lf蛋白的释放,但不影响其mRNA合成。
Objective To investigate the effect of ferric ammonium citrate (FAC) on lactoferrin (Lf) expression in activated microglia induced by lipopolysaccharides (LPS), Methods Primarily cultured rat microglia were respectively treated with FAC,LPS and FAC+LPS. Enzyme-linked immunosorbent assay (ELISA) was employed to examine the concentration of Lf in the cell supernatant, and Lf mRNA was detected using real-time PCR. Results Both the protein and mRNA levels of Lf in LPStreated group obviously increased as compared with the control (F = 458.153,78.884 ;q = 18. 786,10.683 ~P~0.05) ; in FAC-bLPS group, the protein level of Lf was remarkably enhanced (q=12.831, P ~0.05), but Lf mRNA was not elevated compared with LPS group. There was no statistical difference between FAC-treated group and the control with regard to Lf and its mRNA (P〉0.05). Conclusion FAC alone can not increase the expression of Lf in microglia, the release and synthesis of Lf by LPS-activated microglia increase, iron loading enhances the release of Lf in LPS-activated microglia, but does not affect the synthesis of Lf mRNA.