活化蛋白C经胞外信号调节激酶途径抑制肿瘤坏死因子-α介导的炎症反应

Activated protein C ameliorates tumor necrosis factor-α-induced inflammatory response of endothelium Via the extracellular signal-regulated kinase

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 study whether activated protein C（APC） can inhibit TNF-α-induced inflammatory response Via the extracellular signal-regulated kinase（ERK）.Methods Human umbilical vein endothelial cells（HUVECs） were collected and cultured with tumor necrosis factor-α（TNF-α）,normal subjects and TNF-α＋rhAPC group.The protein of ICAM-1,VCAM-1,E-selectin in plasma,ERKmRNA and protein of ERK in HUVECs were detected by enzyme linked immunosorbent assay（ELISA）,reverse transcription polymerase chain reaction（RT-PCR） and western bloting（WB）,respectively.Results The expression of ERKmRNA,phosphor-ERK in PBMCs and the concentration of ICAM-1,VCAM-1,E-selectin in supernatants were higher in TNF-α group than those in normal control group（all P0.05）.By TNF-α＋rhAPC group,the above indexes were decreased significantly compared with those in TNF-α group（all P0.05）.Conclusion Activated protein C ameliorates TNF-α-induced inflammatory response by inhibiting the production of adhesion molecules Via the ERK.