脂多糖诱导的M1型巨噬细胞中Pim-1的动态表达

Expression of Pim-1 in classically activated macrophages induced with lipopolysaccharides

目的:探讨巨噬细胞活化过程中P i m-1的动态表达及抑制磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K),P38丝裂原活化蛋白激酶(P38 mitogen-activated protein kinase,P38MAPK),Janus家族酪氨酸激酶2(Janus kinase 2,JAK2),细胞外信号调节激酶(extracellular regulated protein kinase 1/2,ERK1/2)信号关键分子后Pim-1的表达情况.方法:利用q-RT PCR、Western blot技术检测经脂多糖(lipopolysaccharides,LPS)处理后0、1、2、4、8、12、24 h巨噬细胞中P i m-1m R N A及蛋白的动态表达及P I3K、P38MAPK、JAK2及MEK1/2抑制剂对Pim-1蛋白表达的影响.结果:巨噬细胞中Pim-1的表达随LPS不同作用时间而改变,Pim-1 m RNA表达在LPS刺激后2 h达到高峰,为基础值6倍,12 h后Pim-1 m RNA表达回落至基础水平,LPS刺激1-8 h内Pim-1蛋白表达水平增高,12 h后P i m-1蛋白表达回落至基础水平;P I3K、P38MAPK、JAK2及MEK1/2抑制剂均能下调巨噬细胞中Pim-1蛋白水平.结论:Pim-1 m RNA、Pim-1蛋白的表达是巨噬细胞活化过程中的早期事件;PI3K、P38MAPK、JAK2及MEK1/2抑制剂均影响巨噬细胞中Pim-1的表达,提示Pim-1为相关信号通路的下游分子.

AIM：To explore the expression of Pim-1mRNA and protein during the activation of macrophage and to assess whether inhibition of four key signaling molecules phosphatidylinositol 3-kinase（PI3K）,P38 mitogenactivated protein kinase（P38MAPK）,Janus kinase 2（JAK2）,and ERK kinase（MEK1/2）may influence the expression of Pim-1 protein.METHODS：Pim-1 mRNA and protein expression was examined by real-time PCR and Western blot,respectively,in macrophage cells treated with lipopolysaccharides（LPS）for 0,1,2,4,8,12,and 24 h.After macrophage cells were treated with inhibitor of PI3K（LY294002）,P38MAPK（SB203580）,MEK1/2（AG490） or JAK2（AG490）,LPS induced upregulation of Pim-1 expression was examined by Western blot.RESULTS：Expression of Pim-1 changed with the time of LPS treatment.Pim-1 mRNA and protein expression was rapidly induced after LPS treatment for 1h.Pim-1 mRNA expression reached the peak at 2 h（six-fold of the basal level）,and fell back to the basal level at 12 h.Pim-1 expression was on the constant rise from 1h to 8 h,and dropped to the basic level at 12 h.All inhibitors of the four key signaling molecules down-regulated LPS induced expression of Pim-1 protein.CONCLUSION：Up-regulation of Pim-1 is an early event of classical activation of macrophages.Inhibitors for PI3 K,P38MAPK,JAK2 and MEK1/2 suppress Pim-1 protein expression.Pim-1 may be the downstream signal molecule of PI3 K,P38MAPK,JAK2 and MEK1/2signalling pathway.