TNF-α诱导的人肺动脉内皮细胞GITRL的表达及意义

Expression and significance of GITRL in human pulmonary arterial endothelial cells induced by TNF-α

目的研究糖皮质激素诱导的肿瘤坏死因子受体家族相关蛋白配体(GITRL)在TNF-α诱导的人肺动脉内皮细胞(HPAECs)中的表达及意义。方法原代培养HPAECs,分别加入TNF-α0、5、10、20ng/ml培养2、12、24、48、72h后,采用RT-PCR和Western blot分别检测GITRL mRNA和蛋白表达,ELISA法检测GITRL在细胞上清液中的表达水平。结果 TNF-α呈浓度和时间依赖性地促进GITRL mRNA及其蛋白和上清液GITRL表达水平,其中TNF-α浓度为10ng/ml、培养时间为72h时的表达量最高(P<0.05或P<0.01)。结论 GITRL在TNF-α诱导的HPAECs中表达增加,提示GITR/GITRL可能参与HPAECs的损伤机制,在肺动脉高压的发病过程中起重要作用。

Objective To investigate the expression and significance of glucocorticoid-induced tumor necrosis factor receptor family-related protein ligand（GITRL）in human pulmonary arterial endothelial cells（HPAECs）induced by TNF-α.Methods HPAECs were primarily cultured and stimulated with TNF-α0,5,10,20ng/ml for 2,12,24,48,72 hours.The mRNA and protein expressions of GITRL were detected by RT-PCR and Western blot,respectively.The level of GITRL in the supernatant was measured by ELISA.Results The mRNA and protein expressions of GITRL and level of GITRL in the supernatant were significantly increased by TNF-αstimulation in dose-and time-dependent manners,which were the highest in the concentration of TNF-α 10 ng/ml and incubation time of 72hours（P〈0.05 or P〈0.01）.Conclusion GITRL expression in HPAECs is increased by TNF-αstimulation,indicating that GITR/GITRL pathway may be involved in the injury mechanism of HPAECs and play an important role in the pathogenesis of pulmonary arterial hypertension.