IL-1β suppression of VE-cadherin transcription underlies sepsis-induced inflammatory lung injury
Shiqin Xiong, … , Jalees Rehman, Asrar B. Malik
Shiqin Xiong, … , Jalees Rehman, Asrar B. Malik
Published April 16, 2020
Citation Information: J Clin Invest. 2020;130(7):3684-3698. https://doi.org/10.1172/JCI136908.
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Research Article Inflammation Vascular biology

IL-1β suppression of VE-cadherin transcription underlies sepsis-induced inflammatory lung injury

Abstract

Unchecked inflammation is a hallmark of inflammatory tissue injury in diseases such as acute respiratory distress syndrome (ARDS). Yet the mechanisms of inflammatory lung injury remain largely unknown. Here we showed that bacterial endotoxin lipopolysaccharide (LPS) and cecal ligation and puncture–induced (CLP-induced) polymicrobial sepsis decreased the expression of transcription factor cAMP response element binding (CREB) in lung endothelial cells. We demonstrated that endothelial CREB was crucial for VE-cadherin transcription and the formation of the normal restrictive endothelial adherens junctions. The inflammatory cytokine IL-1β reduced cAMP generation and CREB-mediated transcription of VE-cadherin. Furthermore, endothelial cell–specific deletion of CREB induced lung vascular injury whereas ectopic expression of CREB in the endothelium prevented the injury. We also observed that rolipram, which inhibits type 4 cyclic nucleotide phosphodiesterase–mediated (PDE4-mediated) hydrolysis of cAMP, prevented endotoxemia-induced lung vascular injury since it preserved CREB-mediated VE-cadherin expression. These data demonstrate the fundamental role of the endothelial cAMP-CREB axis in promoting lung vascular integrity and suppressing inflammatory injury. Therefore, strategies aimed at enhancing endothelial CREB-mediated VE-cadherin transcription are potentially useful in preventing sepsis-induced lung vascular injury in ARDS.

Authors

Shiqin Xiong, Zhigang Hong, Long Shuang Huang, Yoshikazu Tsukasaki, Saroj Nepal, Anke Di, Ming Zhong, Wei Wu, Zhiming Ye, Xiaopei Gao, Gadiparthi N. Rao, Dolly Mehta, Jalees Rehman, Asrar B. Malik

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Figure 1

Endotoxin impairs CREB-mediated VE-cadherin expression and promotes lung vascular injury.

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Endotoxin impairs CREB-mediated VE-cadherin expression and promotes lung...
(A–D) C57BL/6J mice (n = 6) were injected intraperitoneally with LPS (12 mg/kg) for 1, 3, and 5 days. Lung vascular permeability was analyzed (A). Western blot detection of protein expression in whole lung lysates (B) and in fresh isolated endothelial cells (C) from LPS-challenged mice. (D) Real-time PCR detection of CREB and VE-cadherin mRNA levels in freshly isolated endothelial cells (n = 3). (E) CREB depletion by siRNA decreases VE-cadherin expression in mLMVECs (n = 3). (F) Overexpression of CREB dose-dependently increases VE-cadherin levels in mLMVECs (n = 3). (G) CREB binds to the mouse VE-cadherin promoter in mLMVECs by ChIP assays. Soluble chromatin was coimmunoprecipitated with anti-CREB or an equal amount of rabbit IgG. DNA purified from starting (1% input) and immunoprecipitated samples was subjected to PCR. The amplified region surrounds each proposed CRE binding element within the mouse VE-cadherin promoter regions. (H) Identification of central CREB CREs enabling VE-cadherin transcription by reporter assay (n = 4). mLMVECs were transiently transfected with the indicated sequential deletion luciferase constructs or pGL3.0 basic control vector together with CREB expression vector, and the internal control p-RL-TK vector. Representative immunoblots and the bar graph quantification are shown. *P < 0.05; **P < 0.01; ***P < 0.001.