Hydrogen Attenuates Septic Lung Injury Via Decreasing TF Expression By Activating Trx1

Objective: Septic lung injury is one of the major causes of death induced by sepsis.However,few effective therapy exists.Hydrogen inhalation has recently been recognized as an effective treatment for inflammatory lung injury,but the mechanism remains unknown.Here,we aimed to reveal how hydrogen attenuated Septic lung injury and provide reference value for the clinical application of hydrogen.Methods:(1)Sepsis lung injury model was established by intraperitoneal injection of endotoxin.Sepsis mice in the treatment group were inhaled 4% hydrogen for 8 hours per day(08:00AM ? 16:00PM),and the 7-days survival rate of each group was observed.(2)The changes of pulmonary edema,pulmonary blood flow and heart rate in each group were observed by ultrasonography.(3)Pulmonary hemorrhage,pulmonary edema and neutrophil infiltration were observed by HE staining,immunohistochemistry and immunofluorescence.(4)The proportion of neutrophils in peripheral blood of mice of each group was measured by flow cytometry.(5)The content of IL-6 in serum of mice was detected by enzyme-linked immunoassay(ELISA).(6)THP-1 and HUVEC-C cells were cultured in vitro and stimulated with LPS to observe the treatment results of hydrogen on mononuclear macrophages and vascular endothelial cells after endotoxin infection.(7)The effects of hydrogen on Trx1,TF and MMP-9 were detected by protein immunoblot,gelatinase spectrum and immunofluorescence co-localization.The expression of Trx1 was blocked by thioredoxin(Trx1)inhibitor PX12,and the anti-inflammatory effect of hydrogen was verified through Trx1.Results: Hydrogen significantly improved the survival rate of mice,reduced edema and hemorrhage of pulmonary,infiltration of neutrophil and IL-6 secretion.Inhalation of hydrogen could decrease TF expression and MMP-9 activity while increase Trx1 expression in lung and blood of sepsis mice.LPS-stimulated THP-1 and HUVEC-C cells in vitro also showed that hydrogen could decrease TF expression and MMP-9activity,which were abolished by Trx1 inhibitor PX12.Conclusions: Hydrogen attenuates Septic lung injury via decreasing TF expression and MMP-9 activity by activating Trx1.Targeting Trx1 by hydrogen may be a potential treatment for septic lung injury.