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

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。

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 den -sity 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 su-pernatants of monocytes were collected from each group ,and the IL-8 expressions of monocytes in every group were detec-ted 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 .