趋化因子FKN对外周血单核细胞NF-κB和TNF-α表达的影响及蛋白激酶C在其中的作用

Effect of Fractalkine on NF-κB and TNF-α expression in peripheral blood monocytes and role of protein kinase C

关键词:  FKN  核因子－κB  肿瘤坏死因子  蛋白激酶C

Key words:  fractalkine  NF-κB  TNF-α  PKC

摘要:

［目的］对单核细胞合成核因子－κB （nuclear factor -κappaB，NF-κB）、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）的影响，探讨趋化因子（fractalkine,FKN）-CX3CR1可能存在的信号转导机制及在动脉粥样硬化形成中的作用，并探讨了蛋白激酶C（protein kinase C，PKC）的介导作用。［方法］①Ficoll密度梯度离心法分离外周血单核细胞。②每份提取的单核细胞分为空白对照组、FKN、RO31-8220（PKC特异性阻断剂）组。③免疫细胞化学法检测各组单核细胞中NF-κB的表达。④酶联免疫法检测各组培养液中TNF-α的表达水平。［结果］FKN、RO31-8220与空白对照组的NF-κB的阳性细胞率分别为（34.80±2.69）%，（20.10±2.78）%与（3.80±0.95）%（三组间差异P<0.05）；TNF－α含量分别为（1506.1±69.3）pg/mL，（820.2±22.8）pg/mL，（80.5±5.5）pg/mL（三组间差异P<0.05）。［结论］FKN－CX3CR1能增加NF-κB、TNF－α单核细胞的表达；而FKN与CX3CR1结合以后，可能通过激活细胞内蛋白激酶C，进而诱导单核细胞合成NF-κB、TNF－α。

Abstract:

［Objective］The effect of Fractalkine( FKN )on the expression of NF-κB 、TNF－αand the role of protein kinase C were investigated.［Methods］①Peripheral blood monocytes were isolated by Ficoll-Paque gradient centrifugation.②The monocytes were divided into : FKN、RO31-8220 and control groups.③The NF-κB expression of the monocytes in each group was detected by immune cytochemsitry.④The level of TNF-α in the supernatant of monocytes from each group was determined by enzyme-linked immunosorbent assay(ELISA). ［Results］In FKN,RO31-8220 and control groups,the expression rates of NF-κB were (34.80±2.69)%,(20.10±2.78)% and (3.80±0.95)% respectively (P<0.05);and levels of TNF-α were (1506.1±69.3)pg/mL,(820.0±22.8)pg/mL and (80.5±5.5)pg/mL respectively (P<0.05) .［Conclusions］FKN－CX3CR1 may increase the expressions of NF-κB and TNF－α in peripheral blood monocytes and FKN initiates intracellular signal conductive mechanism with activating PKC.