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目的观察活化蛋白C(APC)能否经胞外信号调节激酶(ERK)途径抑制肿瘤坏死因子-α(TNF-α)介导的炎症反应。方法分离培养并分成3组:正常对照组、TNF-α组、TNF-α+rhAPC组人脐静脉内皮细胞。分别用ELISA法检测细胞上清液中细胞间黏附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)、E-选择素(E-selectin)浓度;用逆转录聚合酶链反应(RT-PCR)与western bloting技术,检测人脐静脉内皮细胞ERKmRNA、磷酸化ERK蛋白的表达。结果 TNF-α组,细胞上清液中ICAM-1、VCAM-1、E-selectin的浓度较正常对照组显著升高(均P<0.05)。TNF-α+rhAPC组,细胞上清液中ICAM-1、VCAM-1、E-selectin的浓度较TNF-α组显著降低(均P<0.05)。TNF-α组,人脐静脉内皮细胞ERK mRNA、磷酸化蛋白表达水平较正常对照组均显著升高(均P<0.05)。TNF-α+rhAPC组,ERK mRNA、磷酸化蛋白表达水平较TNF-α组均显著降低(均P<0.05)。结论活化蛋白C通过抑制黏附分子的产生来调节TNF-α介导的炎症反应,其作用机制之一可能是通过ERK途径来实现。
Objective To investigate whether activated protein C (APC) inhibits tumor necrosis factor-α (TNF-α) -mediated inflammation through extracellular signal-regulated kinase (ERK) pathway. Methods The cells were divided into 3 groups: normal control group, TNF-α group, TNF-α + rhAPC group, human umbilical vein endothelial cells. The concentrations of ICAM-1, VCAM-1 and E-selectin in the supernatant of the cells were detected by ELISA. The expression of ERK mRNA and phospho-ERK protein in human umbilical vein endothelial cells was detected by RT-PCR and western blotting. Results The concentrations of ICAM-1, VCAM-1 and E-selectin in TNF-α group and cell supernatant were significantly higher than those in normal control group (all P <0.05). The concentrations of ICAM-1, VCAM-1 and E-selectin in TNF-α + rhAPC group and cell supernatant were significantly lower than those in TNF-α group (all P <0.05). The levels of ERK mRNA and phosphorylated protein in human umbilical vein endothelial cells in TNF-α group were significantly higher than those in normal control group (all P <0.05). The levels of ERK mRNA and phosphorylated protein in TNF-α + rhAPC group were significantly lower than those in TNF-α group (all P <0.05). Conclusion Activated protein C regulates TNF-α-mediated inflammation by inhibiting the production of adhesion molecules. One of the mechanisms may be through the ERK pathway.