What question did this study set out to answer?

To investigate the role of hypoxia in acute lung injury and identify potential pharmacological interventions for attenuation.

April 16, 2026Open Access

Hypoxia synergizes cGAS-STING activation and AKT1 phosphorylation to drive pulmonary inflammation in one-lung ventilation: therapeutic attenuation by RU.521 and MK2206

Key Points

To investigate the role of hypoxia in acute lung injury and identify potential pharmacological interventions for attenuation.
Established an OLV model in Sprague-Dawley rats.
Assessed lung tissue damage via histological staining and inflammatory factor quantification using qPCR.
Performed high-throughput transcriptome sequencing to identify differentially expressed genes related to lung injury.
Created a hypoxia reoxygenation model in mouse lung epithelial cells for mechanism validation.
Transcriptomic analysis showed increased gene signatures for DNA damage and oxidative stress.
cGAS-STING pathway was activated with elevated cGAS and AKT1 phosphorylation in lung tissue.
Genetic knockdown of cGAS or AKT1 reduced cGAS-STING pathway signaling, confirming its role in inflammation.
RU.521 and MK2206 effectively reduced cGAS-STING-mediated inflammation and oxidative stress in alveolar cells.

Abstract

Background To explore the mechanisms involved in hypoxia-induced acute lung injury (ALI) and oxidative stress during one-lung ventilation (OLV) in order to find pharmacological interventions that can attenuate OLV injury. Methods An OLV model was established in Sprague-Dawley (SD) rats. Lung tissue damage was assessed via histological staining and by quantifying inflammatory factors (TNF-α, IL-6, IL-1β) using qPCR. High-throughput transcriptome sequencing of left lung tissues from the OLV group and the two-lung ventilation (TLV) group screened for differentially expressed genes (DEGs) in pathways and biological functions associated with lung injury. Furthermore, a hypoxia reoxygenation (H/R) model was established using the mouse lung epithelial cell line (MLE-12) for further mechanism mapping and validation in vitro. Results Transcriptomic analysis revealed upregulated gene signatures linked to DNA damage, mitochondrial membrane permeabilization, oxidative stress, and innate immune activation. KEGG enrichment identified innate immune pathways, specifically cytosolic DNA sensing. Subsequent in vivo and in vitro studies confirmed cGAS-STING pathway activation, characterized by increased expression and enhanced phosphorylation of downstream components. Genetic knockdown of cGAS or AKT1 (a kinase required for downstream phosphorylation of key cGAS-STING downstream effectors) suppressed cGAS-STING pathway signaling. Conclusion This study highlights that innate immunity contributes to lung tissue damage during OLV. Hypoxia-induced DNA damage activates the cytoplasmic DNA-sensing cGAS-STING pathway. Subsequently, AKT1 modulates the downstream phosphorylation of TBK1, IRF3, and NF-κB, thereby promoting DNA damage-associated inflammation in alveolar epithelial cells. Pharmacologically, both the cGAS inhibitor RU.521 and AKT1 inhibitor MK2206 attenuated cGAS-STING-mediated inflammation and mitigated hypoxia-induced oxidative stress in alveolar epithelia. Clinical trial number Not applicable.

Hypoxia synergizes cGAS-STING activation and AKT1 phosphorylation to drive pulmonary inflammation in one-lung ventilation: therapeutic attenuation by RU.521 and MK2206

Key Points

Abstract

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