趋化因子FKN对人外周血单个核细胞IL-8表达的影响及ERK、PI3K和NF-κB在其中的作用

Effect of chemotatic factor FKN on IL-8 expression of monocytes in peripheral blood and the functions of ERK、PI3K and NF-κB

关键词:  FKN  白细胞介素-8  细胞外调节  磷脂酰肌醇-3激酶  核因子-κB

Key words:  [Key words] 　fractalkine  interleukin-8  extracellular regulated protein kinases  phosphatidylinositol 3-kinase  nuclear factor-κB

摘要:

目的　通过研究趋化因子Fractalkine（FKN）对人外周血单个核细胞IL-8表达的影响，以及信号分子细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)、磷脂酰肌醇-3激酶（phosphatidylinositol 3-kinase，PI3K）、核因子-κB（nuclear factor-κB，NF-κB）在其中的作用,探讨FKN促进动脉粥样硬化（atherosclerosis，AS）的机制。方法　利用密度梯度离心法分离健康人外周血单个核细胞；将提取的单个核细胞分为：空白对照组、FKN组、PD98059（ERK阻断剂）组、LY294002（PI3K阻断剂）组、PDTC（NF-κB阻断剂）组、FKN+PD98059组、FKN+LY294002组、FKN+PDTC组；分别于培养12 h和24 h后收集细胞培养上清，应用ELISA检测各组单个核细胞中IL-8的表达情况。结果　(1)细胞培养12 h或24 h后，PD98059组、LY294002组和PDTC组与空白组比较，IL-8表达有减少趋势，但差异无显著性意义（P>0.05）。(2)FKN组较空白组IL-8表达明显减少（P<0.05），FKN+PD98059组和FKN+LY294002组较FKN组IL-8表达明显增加（P<0.05），FKN+PDTC组较FKN组、FKN+PD98059组和FKN+LY294002组IL-8表达均明显减少（P<0.05）。(3)细胞培养24 h后，各组IL-8表达均较12 h明显减少(P<0.05)。结论　趋化因子FKN可能通过ERK、PI3K信号途径抑制单个核细胞IL-8的表达，而通过NF-κB途径促进IL-8的表达，FKN对单个核细胞IL-8的表达具有双向调节作用，但以抑制作用为主。

Abstract:

[Abstract] Objective　To observe the effect of chemotatic factor FKN on IL-8 expression of monocytes in peripheral blood and the function of extracellular regulated protein kinases(ERK),phosphatidylinositol 3-kinase(PI3K) and nuclear factor κB(NF-κB). Methods　Peripheral blood monocytes were isolated from fresh blood of healthy volunteers by the density gradient centrifugation. The isolated monocytes were divided into eight groups: control, FKN, PD98059, LY294002, PDTC (NF-κB inhibitor), FKN+PD98059, FKN+LY294002, FKN+PDTC. At 12 h and 24 h, respectively, the supernatants of monocytes were collected from each group,and the IL-8 expressions of monocytes in every group were detected with ELISA. Results　The expressions of IL-8 in PD98059, LY294002 and PDTC group showed a decreasing trend, but not statistically significant (P>0.05),compared with that of the control and the expressions of IL-8 in FKN group was decreased compared with that of control（P<0.05）.The expression of IL-8 from FKN+PD98059 and FKN+LY294002 group were increased compared with that from FKN group（P<0.05）. The expression of IL-8 from FKN+PDTC was significantly decreased compared with those from FKN、FKN+PD98059 and FKN+LY294002（P<0.05）. Cells cultured for 24 h group,IL-8 expression in each group was significantly decreased compared with 12 h (P<0.05). Conclusion　Chemokine factor FKN could inhibit IL-8 expression in mononuclear cells through the ERK、PI3K pathway, promote the expression of IL-8 by NF-κB pathway.