Experimental design

TAL efficacy was verified in 2 lung injury models with different severities induced by changing the dose of LPS (E. coli O127: B8; Sigma-Aldrich Co. LLC, St. Louis, MO, USA). A severe acute lung injury model was generated using 5 mg/kg of LPS in experiment 1, and 10 mg/kg of LPS was used to create a lethal acute lung injury model in experiment 2 (Supplemental Figure ^S1).

In experiment 1, 15 rats were randomly divided into 3 groups: 5 rats in the TAL treatment group received 5 min of TAL and 3 h of MGV 20 min after LPS administration (LPS + TAL group); five rats in the MGV treatment group received 5 min and 3 h of MGV 20 min after LPS administration (LPS + MGV group), and 5 rats in the sham group received 5 min and 3 h of MGV 20 min after PBS administration instead of LPS (PBS + MGV group). After 2 days, the haemodynamic and blood gas parameters were measured. The lungs were removed from the chest, and BAL and histopathological analyses were performed.

In experiment 2, 10 rats were randomly divided into 2 groups of 5 rats each: the TAL treatment group (LPS + TAL group) and the MGV treatment group (LPS + MGV group). The treatments were the same as in experiment 1, and all surviving rats were sacrificed after 7 days. The rats’ lungs were subsequently isolated for histopathological analyses. The primary outcome of this experiment was the survival rate of the rats.