NRF2/miR-17-5p/TMEM16F axis regulates the crosstalk of inflammation and thrombosis in sepsis.

Abstract

BACKGROUND: Sepsis is characterized by the simultaneous activation of inflammation and coagulation in response to systemic infection, leading to microvascular dysfunction and consequential multiorgan failure. Previous studies have revealed that TMEM16F is involved in blood coagulation and inflammatory response. However, its detailed regulatory molecular mechanisms and functions in sepsis remain unknown. OBJECTIVES: In this study, we aim to investigate the regulatory effects and the underlying mechanism of TMEM16F in sepsis-induced microvascular thrombosis and inflammation. METHODS: We used septic patients, lipopolysaccharide (LPS)-induced septic mouse models, and in vitro cellular experiments with LPS-stimulated endothelial cells. Bioinformatical analysis and molecular biological techniques were performed to identify the target genes of TMEM16F. RESULTS AND CONCLUSIONS: We found that TMEM16F was highly expressed in vivo and in vitro septic model, and its deletion decreased mortality, inflammation, and the microthrombi formations in LPS-induced septic mice. Additionally, silencing NRF2 not only inhibited TMEM16F expression but also improved LPS-induced thrombosis, inflammation, and organ injury, which could be reversed by inhibition of miR-17-5p. By bioinformatics analysis with subsequent chromatin immunoprecipitation-polymerase chain reaction and luciferase activity experiments, we revealed that the transcription factor NRF2 drove TMEM16F transcription through promoter binding while suppressing miR-17-5p, thereby increasing TMEM16F expression in endothelial cells. Furthermore, plasma concentrations of TMEM16F and miR-17-5p were correlated with coagulation activation, inflammation, and disseminated intravascular coagulation scores in septic patients. Overall, these findings identify the NRF2/miR-17-5p/TMEM16F axis may be a potential therapeutic target for sepsis.