IKKβ regulates the expression of coagulation and fibrinolysis factors through the NF‑κB canonical pathway in LPS‑stimulated alveolar epithelial cells type II

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

doi:10.3892/etm.2019.7928

IKKβ regulates the expression of coagulation and fibrinolysis factors through the NF‑κB canonical pathway in LPS‑stimulated alveolar epithelial cells type II

Authors:
- Bo Liu
- Yahui Wang
- Yanqi Wu
- Yumei Cheng
- Hong Qian
- Huilin Yang
- 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: August 20, 2019 https://doi.org/10.3892/etm.2019.7928
Pages: 2859-2866
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Aim: Hypercoagulation and fibrinolysis inhibition in the alveolar cavity are important characteristics in acute respiratory distress syndrome (ARDS). Alveolar epithelial cells type Ⅱ (AEC II) have been confirmed to have significant role in regulating alveolar hypercoagulation and fibrinolysis inhibition, but the mechanism is unknown. Nuclear factor‑κB (NF‑κB) signaling pathway has been demonstrated to participate in the pathogenesis of these two abnormalities in ARDS. The purpose of the present study is to explore whether controlling the upstream crucial factor IκB kinase (IKK)β could regulate coagulation and fibrinolysis factors in LPS‑stimulated AEC II. Materials and methods: An IKKβ gene regulation model (IKKβ+/+ and IKKβ‑/‑) was prepared using lentiviral vector transfection. The models with wild type cells were all stimulated by lipopolysaccharide (LPS) or saline for 24 h. Expression of the related proteins were determined by western‑blotting, ELISA and revere transcription‑PCR respectively. Tissue factor (TF) procoagulant activity and nuclear p65 protein level were also detected. Results: IKKβ increased in IKKβ+/+ cells but decreased in IKKβ‑/‑ cells. LPS stimulation promoted the expression of p‑IκBα, p65, p‑p65 and p‑IKKβ as well as TF and plasminogen activator inhibitor (PAI)‑1, at the mRNA or protein level, and this was significantly enhanced by IKKβ upregulation but weakened by IKKβ downregulation. TF procoagulant activity presented the same changes as the molecules above. ELISAs showed additional increases in the concentrations of as thrombin antithrombin, procollagen Ⅲ propeptide, thrombomodulin and PAI‑1 in IKKβ+/+ cell supernatant under LPS stimulation, however they decreased in IKKβ‑/‑. The level of as antithrombin Ⅲ however, appeared to show the opposite change to those other factors. Immunofluorescence demonstrated a greatly enhanced expression of p65 in the nucleus by IKKβ upregulation, which was reduced by IKKβ downregulation. Conclusions: IKKβ could regulate the expression and secretion of coagulation and fibrinolysis factors in LPS‑stimulated AEC II via the NF‑κB p65 signaling pathway. The IKKβ molecule is expected to be a new target for prevention of coagulation and fibrinolysis abnormalities in ARDS.

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