细胞外信号调节激酶信号通路对小鼠脾脏树突状细胞表型和功能的影响

Effect of extracellular eignal-regulated kinase pathway on the phenotype and function of mouse splenic dendritic cell

目的探讨细胞外信号调节激酶(ERK)信号通路对小鼠脾脏树突状细胞(DC)表型和功能的影响。方法应用免疫磁珠细胞分选方法分离小鼠脾脏DC,流式细胞术鉴定细胞纯度。应用U1026抑制ERK信号通路。流式细胞术检测DC表面共刺激分子的表达及细胞凋亡;酶联免疫吸附测定(ELISA)检测DC细胞因子的分泌。结果U1026处理组较对照组DC表面CD86(P<0.01)和CD40(P<0.05)的表达下调。脂多糖(LPS)+U1026组较LPS组DC表面CD80(P<0.01)和CD86(P<0.05)表达下调。对照组DC凋亡比例为(57.09±0.64)%,U1026处理组DC凋亡比例为(76.65±0.25)%,2组DC凋亡比例比较差异有统计学意义(P<0.01)。对照组肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)及转化生长因子-β(TGF-β)水平分别为(78.22±5.71)、(47.75±0.77)、(57.72±2.34)mg·L-1,U1026处理组TNF-α、IL-6及TGF-β水平分别为(22.22±0.32)、(23.35±0.32)、(34.21±1.76)mg·L-1,2组各细胞因子水平比较差异均有统计学意义(P<0.05,P<0.01)。结论 ERK信号通路可调控DC表面共刺激分子的表达、细胞凋亡及细胞因子的分泌。

Objective To investigate the effect of extracellular eignal-regulated kinase（ ERK） pathway on the phenotype and function of mouse splenic dendritic cell（ DC）. Methods Magnetic bead cell sorting was used to isolate splenic DC,and then the flow cytometry was used to analyze the purity of DC. U1026 was used to inhibit the activity of ERK pathway. The expression of DC costimulatory molecules and apoptosis were analyzed by flow cytometry. The secretion of cytokines of DC were detected by enzyme-linked immunosorbent assay. Results The expressions of CD86 and CD40 of DC in U1026 treatment group were less than those in control group（ P〈0. 01,P〈0.05）; the expressions of CD80 and CD86 of DC in lipoplysaccha rides（ LPS） ＋ U1026 group were less than those in LPS group（ P〈0.01,P〈0.05）. The proportion of DC apoptosis in control group and U1026 treatment group was（ 57. 09 ± 0. 64） % and（ 76. 65 ± 0. 25） % respectively,there was statistic difference of the proportion of DC apoptosis between the two groups（ P〈0.01）. The levels of tumor necrosis factor-α（ TNF-α）,interleukin-6（ IL-6） and transforming growth factor-β（ TGF-β） in control group was（ 78. 22 ± 5. 71）,（ 47. 75 ± 0. 77）,（ 57. 72 ±2. 34） mg·L- 1respectively; the levels of TNF-α,IL-6 and TGF-β in U1026 treatment group was（ 22. 22 ± 0. 32）,（ 23. 35 ±0. 32）,（ 34. 21 ± 1. 76） mg·L- 1respectively; there were statistic difference of the levels of TNF-α,IL-6 and TGF-β between the two groups（ P〈0.05,P〈0.01）. Conclusion ERK pathway can regulates the costimulatory molecules expression,apoptosis and cytokine secretion of spleenic DC.