核因子κB在烧伤血清诱导内皮细胞细胞间黏附分子1表达中的作用

目的探讨核因子κB(NF-κB)在烧伤血清诱导内皮细胞(EC)分泌细胞间黏附分子(ICAM)1中的作用。方法人脐静脉内皮细胞(HUVEC)培养后,分别用正常人血清(对照组)、烧伤患者血清(烧伤血清组)、吡咯烷二硫代氨基甲酸盐(PDTC)+烧伤患者血清(PDTC组)刺激。于刺激0.5、1.0、2.0、4．0 b时采用电泳迁移率分析法测定HUVEC NF-κB的活性;流式细胞术检测刺激3．0、6.0、12．0、24．0 h时HUVEC膜表面ICAM-1的表达。结果刺激后烧伤血清组、PDTC组细胞NF-κB活性均明显高于对照组(P<0．01),1．0 h时达峰值(21.03±4．87)、(7.44±0．60)×104积分灰度值],以后逐渐降低;PDTC组明显低于烧伤血清组(P<0.01)。刺激后烧伤血清组、PDTC组ICAM-1的表达均增加,刺激12．0 h吋达峰值(平均荧光密度各为327±37、142±31),与对照组比较,差异有统计学意义(P<0．01);PDTC组刺激12．0、24．0 h时低于烧伤血清组(P<0．01)。结论烧伤血清通过活化NF-κB,从而启动EC对黏附分子的合成和释放,提示NF-κB在烧伤血清诱导EC分泌黏附分子过程中起重要作用。

The role of nuclear factor-κB in the burn serum-induced expression of intercellular adhesion molecules-1 in endothelial cells

OBJECTIVE: To explore the role of nuclear factor-kappaB (NF-kappaB) in the burn serum induced expression of intercellular adhesion molecules-1 (ICAM-1) in endothelial cells. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and divided into control (with normal serum stimulation), burn serum (B, with burn serum stimulation) and PDTC (with burn serum and PDTC stimulation) groups. The NF-kappaB activity in endothelial cells was determined with electrophoretic mobility shift assay (EMSA) at 0.5, 1.0, 2.0, 4.0 post-stimulation hour (PSH). The expression of ICAM-1 at 3.0, 6.0, 12. 0, 24.0 PSH was detected by flow cytometry. RESULTS: The NF-kappaB activity in endothelial cells in burn serum group and PDTC group were markedly higher than that in control group (P <0.01), and it reached the peak at 1.0 PSH (21.03 +/- 4.87), (7.44 +/- 0.60) x 10(4) A], respectively, then gradually decreased. But it was obviously lower in PDTC group than that in burn serum group ( P <0. 01 ). The expression of ICAM-1 was gradually increased in both burn serum group and PDTC group, reaching the peak level at 12.0 PSH (327 +/- 37), (142 +/- 31) mean fluorescence intensity], respectively, which were significantly higher than that in control group (P <0.01). But it was evidently lower in PDTC group than that in burn serum group at 12.0 and 24.0 PSH (P <0.01). CONCLUSION: Burn serum can initiate the synthesis and release of adhesion molecules in endothelial cells through activation of NF-kappaB, indicating that NF-kappaB plays an important role in the process of burn serum induced endothelial secretion of adhesion molecules.