Septic serum mediates inflammatory injury in human umbilical vein endothelial cells via reactive oxygen species, mitogen activated protein kinases and nuclear factor- [kappa]B

• Published in: International journal of molecular medicine 2021-01, Vol.47 (1), p.267
• Main Authors: Xu, Shouzhu, Yan, Yu, Yan, Zhijiao, Xu, Jie, Qi, Baoning, Li, Juan, Zhang, Zhigang, Han, Yuanping, Zhao, Jing
• Format: Article
• Language: English
• Subjects: Acetylcysteine; Analysis; Endothelium; Enzyme-linked immunosorbent assay; Fluorescence microscopy; Infection; Interleukins; Mitogens; Protein kinases; Superoxide
• ISSN: 1107-3756
• DOI: 10.3892/ijmm.2020.4785

Sepsis-induced blood vessel dysfunction is mainly caused by microvascular endothelial cell injury. However, the mechanism underlying sepsis-induced endothelial cell injury remains unclear. The present study hypothesized that sepsis-induced inflammatory injury of endothelial cells may be the first step of endothelial barrier dysfunction. Therefore, the present study aimed to uncover the mechanism underlying the inflammatory effects of sepsis. A rat model of cecal ligation and puncture-induced sepsis was established, and septic serum was collected. Subsequently, human umbilical vein endothelial cells (HUVEcs) were treated with the isolated septic or normal serum. HUVEC viability was assessed using a Cell Count Kit-8 assay. Furthermore, transmission electron microscopy and reverse transcription-quantitative PCR (RT-qPCR) analysis were carried out to observe the cell morphology and determine the mRNA expression levels in septic serum-induced HUVECs. The protein expression levels were evaluated by western blot analysis, and the secretion of the inflammatory factors interleukin (IL)-1[beta], IL-6 and tumor necrosis factor (TNF)-[alpha] was determined by ELISA. Additionally, reactive oxygen species (ROS) generation and nuclear factor (NF)-[kappa]B nuclear translocation were observed under a fluorescence microscope. The results of the present study demonstrated that HUVEC viability was significantly decreased following 12- or 24-h treatment with septic serum. In addition, chromatin condensation, mitochondrial vacuolization and endoplasmic reticulum degranulation were observed following treatment with septic serum. Furthermore, the secretion levels of IL-1[beta], IL-6 and TNF-[alpha] were increased in septic serum-stimulated HUVECs. Septic serum treatment also enhanced superoxide anion generation, promoted extracellular signal regulated kinase 1/2 (ERK1/2), N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38) phosphorylation, and increased NF-[kappa]B levels in the nuclei of HUVECs. Finally, pre-treatment of HUVEcs with the antioxidant N-acetylcysteine, the ERK1/2 inhibitor PD98059, the p38 inhibitor SB203580, the JNK inhibitor SP610025 or the NF-[kappa]B inhibitor pyrrolidine dithiocarbamate restored the septic serum-induced IL-1[beta], IL-6 and TNF-[alpha] expression. In conclusion, the results of the current study suggested that the septic serum-induced endothelial cell injury may be mediated by increasing ROS generation, activation of mitogen-