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 2

IL-1β represses cAMP-CREB signaling.

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IL-1β represses cAMP-CREB signaling. (A) mLMVECs were treated with IL-1β...
(A) mLMVECs were treated with IL-1β (5 ng/mL) or vehicle for the indicated time. Transwell permeability to HRP was measured (n = 6). (B and C) mLMVECs were stimulated with increasing doses of IL-1β for 24 hours (B) or stimulated with IL-1β (5 ng/mL) over time (C). Protein expression was detected by Western blot. (D–G) mLMVECs were stimulated with IL-1β (5 ng/mL) over time. RNA expression levels were analyzed by RT-PCR (D). Ubiquitination status of CREB was analyzed by immunoprecipitation with CREB antibody and blotting with an anti-ubiquitin antibody (E). sAC activity was detected (F) (n = 4). Intracellular cAMP levels were measured (G) (n = 4). (H) CREB itself binds to the putative CREs within the CREB promoter in mLMVECs by ChIP assays. The amplified region surrounds each proposed CRE binding element within the mouse CREB promoter regions. (I) mLMVECs were stimulated with IL-1β (5 ng/mL) for the indicated time. Effect of IL-1β treatment on CREB binding to the promoter regions (CRE2 and CRE6) in mLMVECs was investigated by ChIP assays (n = 3). *P < 0.05; **P < 0.01; ***P < 0.001; NS, no significant difference. Statistics obtained from ANOVA.