Study On The Protective Effect Of 4% Hydrogen Inhalation On Lung Injury In Septic Mice By Regulating The Release Of NETs

Objective:to explore whether 4%hydrogen inhalation therapy can protect the lung injury of septic mice by regulating the release of NETs in the lung tissue of CLP septic mice.Methods:male adult C57BL/6 mice,weighing 20-25 g,8-10 weeks,were randomly divided into four groups:normal group(NC),sham operation group(Sham),sepsis model group(CLP),sepsis model+ 4%hydrogen inhalation group(CLP+H2).Sepsis mouse model was established by cecal ligation and puncture(CLP).Observe the 7-day survival rate of four groups of mice.At 24 h and 48 h after sham operation or CLP,eyeball and lung tissue were removed,blood routine was detected by automatic instrument,the concentration of ALT,AST,Crea,Urea in serum and SOD activity in lung tissue were detected by colorimetry,IL-6,IL-10 and MPO in plasma and lung tissue were detected by Elisa method,lung tissue was detected by HE staining and NETs immunofluorescence,and the expression of Cit-H3 and NE in lung tissue was detected by Westernblot method.Results:the 7-day survival rate of mice in NC group was 100%,94.44%and 66.67%in NC group,94.44%and 83.33%in H2 group.Compared with NC group,the survival rate of CLP group and CLP+H2 group was significantly lower,and the clinical score of 24-hour sepsis was significantly higher in CLP+H2 group than that of CLP group(P<0.05).Compared with CLP group,the 7-day survival rate of CLP+H2 group was significantly higher and the clinical score of 24-hour sepsis was lower(P<0.05).Compared with NC group,white blood cells,neutrophils and lymphocytes in CLP group and CLP+H2 group were significantly lower,serum ALT,AST,Crea and Urea concentrations were significantly higher,plasma IL-10 and IL-6 were higher,lung tissue pathological score was significantly higher,lung tissue IL-10 and IL-6 were higher,and lung tissue SOD activity was decreased.The expression of MPO in lung tissue increased.Compared with CLP group,lymphocytes in CLP+H2 group increased at 48h,leukocytes and neutrophils had no significant difference,serum AST,ALT,Urea decreased,serum Crea had no significant difference,plasma IL-10 increased,IL-6 had no significant difference,and lung pathological score decreased.IL-6 in lung tissue increased slightly,but there was no difference(P>0.05),and IL-10 was not clear.The activity of SOD in lung tissue increased and the expression of MPO in lung tissue decreased.Compared with NC group,the expression of NETs-related protein Cit-H3 and NETs immunofluorescence in lung tissue of CLP group and CLP+H2 group increased,while the expression of NETs-related protein Cit-H3 and NE decreased significantly in CLP+H2 group compared with CLP group(P<0.05).),NETs immunofluorescence quantitative decreased significantly(P<0.05).Conclusion:1.4%hydrogen inhalation can protect CLP septic mice,improve their survival rate,reduce organ function injury 2.4%hydrogen inhalation can reduce oxidative stress in lung tissue of CLP septic mice,and increase antioxidant enzyme activity 3.4%hydrogen inhalation can reduce the release of NETs in lung tissue of CLP septic mice and protect septic lung injury...