We aimed to investigate and compare the effects of rapid (NaHS) and slow (GYY4137 and AP39) hydrogen sulfide (H2S) releasing donors on LPS-induced tracheal hyperreactivity and pro-inflammatory cytokine levels in lung tissues of mice. Tissues were isolated from male BALB/c mice and incubated with LPS (10 mu g/mL) in tissue culture. The subgroups were incubated with NaHS, GYY4137 and mitochondria-targeted donor AP39. LPS incubation did not alter contraction response to carbachol, but enhanced 5-HT and bradykinin-induced contractions in tracheal rings, and elevated IL-1 beta, IL-6 and TNF-alpha levels in lung homogenates. NaHS at 300 mu mol/L and 1000 mu mol/L, GYY4137 at 30 mu mol/L and 100 mu mol/L, and AP39 at 30 nmol/L concentrations inhibited the tracheal hyperreactivity to 5-HT, whereas none of these donors affected the enhanced contraction to bradykinin. GYY4137 was also effective to inhibit 5-HT hyperreactivity acutely. In lung tissues, NaHS prevented the elevation of IL-1 beta level at 1000 mu mol/L, and IL-6 and TNF-alpha levels at 100 mu mol/L concentrations. Incubation with GYY4137 (100 mu mol/L) and AP39 (30 nmol/L and 300 nmol/L) inhibited the increase in IL-6 and TNF-alpha levels, but not IL-1 beta at concentrations that they affected tracheal hyperreactivity. These results indicate that H2S donors can decrease inflammation and prevent airway hyperreactivity.