Long Ding, Yang Junhui, Wu Xiaoling, Gui Yun, Yu Li
Intensive Care Unit, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology Wuhan, China.
Int J Clin Exp Pathol. 2019 Jan 1;12(1):77-86. eCollection 2019.
The aim of this study was to investigate the effect of urokinase-type plasminogen activator (uPA) on cell viability, apoptosis, and inflammatory cytokine levels in septic human umbilical vein endothelial cells (HUVECs). Lipopolysaccharides (LPS) were added to construct septic HUVECs, then the septic HUVECs were treated by uPA, and cell viability, apoptosis, TNF-α, IL-6, GMCSF and uPAR levels were evaluated by CCK-8, AV/PI, qPCR, western blot and ELISA, respectively. Subsequently, uPA shRNA was transferred into septic HUVECs, and the cells viability, cell apoptosis and the expressions of TNF-α, IL-6, GMCSF, as well as uPAR were assessed by the same methods. uPA promoted viability while reducingapoptosis in septic HUVECs. However, uPA had no effect on the regulation of TNF-α or IL-6 expression in septic HUVECs. In addition, uPA elevated the expressions of GMCSF and uPAR in septic HUVECs. After the transfection of uPA shRNA, cell viability was decreased, apoptosis was enhanced, and GMCSF and uPAR expressions were reduced, while TNF-α or IL-6 expression did not vary in septic HUVECs. In conclusion, uPA promotes cell viability, represses apoptosis,and has no effect on regulating inflammatory cytokines in septic HUVECs.
本研究旨在探讨尿激酶型纤溶酶原激活剂(uPA)对脓毒症人脐静脉内皮细胞(HUVECs)的细胞活力、凋亡及炎性细胞因子水平的影响。加入脂多糖(LPS)构建脓毒症HUVECs模型,然后用uPA处理脓毒症HUVECs,分别采用CCK-8法、AV/PI法、qPCR法、蛋白质印迹法及酶联免疫吸附测定(ELISA)法评估细胞活力、凋亡情况、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、粒细胞巨噬细胞集落刺激因子(GMCSF)及uPA受体(uPAR)水平。随后,将uPA短发夹RNA(shRNA)转染至脓毒症HUVECs中,采用相同方法评估细胞活力、细胞凋亡以及TNF-α、IL-6、GMCSF和uPAR的表达。uPA可提高脓毒症HUVECs的活力并减少其凋亡。然而,uPA对脓毒症HUVECs中TNF-α或IL-6的表达调控并无影响。此外,uPA可提高脓毒症HUVECs中GMCSF和uPAR的表达。转染uPA shRNA后,脓毒症HUVECs的细胞活力降低,凋亡增加,GMCSF和uPAR的表达减少,而TNF-α或IL-6的表达无变化。总之,uPA可促进脓毒症HUVECs的细胞活力,抑制其凋亡,且对调节炎性细胞因子无作用。