BAY11‑7082 inhibits the expression of tissue factor and plasminogen activator inhibitor‑1 in type‑II alveolar epithelial cells following TNF‑α stimulation via the NF‑κB pathway

Cheng,Yumei; Liu,Bo; Qian,Hong; Yang,Huilin; Wang,Yahui; Wu,Yanqi; Shen,Feng

doi:10.3892/etm.2020.9608

BAY11‑7082 inhibits the expression of tissue factor and plasminogen activator inhibitor‑1 in type‑II alveolar epithelial cells following TNF‑α stimulation via the NF‑κB pathway

Authors:
- Yumei Cheng
- Bo Liu
- Hong Qian
- Huilin Yang
- Yahui Wang
- Yanqi Wu
- Feng Shen
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Affiliations: Department of Critical Care Medicine, Guizhou Medical University Affiliated Hospital, Guiyang, Guizhou 550001, P.R. China
Published online on: December 28, 2020 https://doi.org/10.3892/etm.2020.9608
Article Number: 177

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Abstract

Pulmonary inflammation strongly promotes alveolar hypercoagulation and fibrinolytic inhibition. NF‑κB signaling regulates the expression of molecules associated with coagulation and fibrinolytic inhibition in type‑II alveolar epithelial cells (AECII) stimulated by lipopolysaccharide. However, whether TNF‑α‑induced alveolar hypercoagulation and fibrinolysis inhibition is also associated with the NF‑κB pathway remains to be determined. The aim of the present study was to determine whether BAY11‑7082, an inhibitor of the NF‑κB pathway, inhibits the expressions of tissue factor (TF) and plasminogen activator inhibitor‑1 (PAI‑1) in AECⅡ in response to TNF‑α. Rat AECII were treated with BAY11‑7082 for 24 h and stimulated with TNF‑α for 1 h. The expression of TF and PAI‑1 were determined using western blotting and reverse transcription‑quantitative PCR. The concentrations of TF and PAI‑1 in culture supernatant were also measured by ELISA. Moreover, levels of NF‑κB p65 (p65), phosphorylated (p)‑p65 (p‑p65), inhibitor of NF‑κB α (IκBα) and p‑IκBα were also evaluated. Immunofluorescence was used to detect p65 levels in cell nuclei. TNF‑α significantly promoted TF and PAI‑1 expression either at the mRNA or protein level in AECII cells. Concentrations of TF and PAI‑1 in supernatant also significantly increased upon TNF‑α stimulation. Furthermore, TNF‑α upregulated the levels of p‑IκBα, p65, and p‑p65 in the cytoplasm. Immunofluorescence analysis indicated that TNF‑α increased p65 translocation from the cytoplasm to the nucleus. However, AECII pre‑treated with BAY11‑7082 expressed lower levels of TF and PAI‑1 following TNF‑α treatment. Levels of p‑IκBα, p65 and p‑p65 in the cytoplasm also decreased, and translocation of p65 from cytoplasm into the nucleus was inhibited by BAY11‑7082 pretreatment. These findings suggest that BAY11‑7082 improves the hypercoagulation and fibrinolytic inhibition induced by TNF‑α in alveolar epithelial cells via the NF‑κB signaling pathway. BAY11‑7082 might represent a therapeutic option for alveolar hypercoagulation and fibrinolytic inhibition in acute respiratory distress syndrome.

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