不同浓度脂多糖对J774A.1细胞CD14表达的影响

Effect of LPS with different doses on CD14 surface expression in J774A.1 cell line

目的:探讨不同时间和不同浓度脂多糖(LPS)对小鼠巨噬细胞株J774A.1细胞CD14表达的影响。方法:用不同浓度LPS刺激J774A.1细胞后,FITC-CD14单克隆抗体染色45 min,用流式细胞术检测J774A.1细胞CD14的表达,观察用LPS刺激不同时间(1、2、4、8及16 h)后J774A.1细胞CD14表达的变化;选择最佳时间后,观察不同浓度(1、10、50、100、500及1 000μg.L-1)LPS对J774A.1细胞CD14表达的变化。结果:在时间-效应关系上,当LPS浓度为1μg.L-1时,刺激1、2及4 h后CD14表达增强(分别为23.80±5.07、23.04±2.88、28.22±1.54),与对照组(12.50±3.71)比较差异有显著性(P<0.01),LPS浓度为10μg.L-1时,刺激1和2 h后CD14表达增强(分别为40.85±6.05、26.63±6.17),与对照组(12.50±3.71)比较差异有显著性(P<0.05,P<0.01),而LPS浓度为50μg.L-1时,刺激1 h后CD14表达增强(47.40±2.85),与对照组(12.50±3.71)比较差异有显著性(P<0.01)。在剂量-效应关系上,刺激4 h时LPS浓度为小于10μg.L-1时CD14表达蛋白明显高于对照组(P<0.01)。结论:用LPS刺激J774A.1细胞表达CD14时,LPS刺激时间最好是在4 h以内,刺激浓度最好不要超过50μg.L-1。

Objective To study the dose- and time-effect relationship between lipopolysaccharide （LPS） and CD14 expression in the cell line J774A. 1 of mouse macrophages. Methods The cell line J774A. 1 was stimulated with different doses of LPS, and the CD14 surface-positive cells from the cell line J774A. 1 were detected by flow cytometry （FCM） 45 rain after stained with FITC-CD14 monoclonal antibody. The changes of CD14 surface expression in the cell line J774A. 1 at different time （1, 2, 4, 8 and 16 h） after stimulated with LPS were observed. After the best stimulant time was selected, the cell line J774A. 1 was treated with different doses （1, 10, 50, 100, 500 and 1 000 μg·L^-1） of LPS and the changes of CD14 surface expression were observed. Results In view of time-effect, compared with control group （the positive rate was 12.50 ± 3.71）, LPS significantly enhanced the CD14 expression at 1, 2 and 4 h after stimulation （the positive rates were 23.80±5.07, 23.04±2.88 and 28. 22± 1.54, respectively） when the dose of LPS was 1μg·L^-1 （P〈0.01）. When the concentration of LPS was 10 μg·L^-1 , CD14 expression strengthened at 1 and 2 h after stimulation （the positive rates were 40. 85±6.05 and 26.63±6.17, respectively） （P〈0.05, P〈0. 01）. LPS obviously stimulated CD14 expression at 1 h after stimulation （the positive rate was 47. 40±2.85） when the dose of LPS was 50 μg·L^-1 （P~〈0.01）. In the dose-effect, the stimulant effect of LPS occurred when LPS was administered only below the dose of 10 μg·L^-1 at 4 h after stimulation （P〈0.01）. Conclusion LPS can significantly stimulate the expression of CD14 in the cell line J774A. 1 below the dose of 50 μg·L^-1 within 4 h.