Objectives Hydrogen sulfide (H2S) is involved in regulating cell apoptosis and proliferation. However, The effects and mechanism of H2S on the apoptosis of mammary epithelial cells that suffer from an inflammatory response remain unknown. Results An inflammatory cell model was used to explore whether exogenous H2S regulates lipopolysaccharides (LPS)-induced cell proliferation and apoptosis. We found that H2S affected cell viability, the inflammatory response and apoptosis in LPS-treated cells in a concentration-dependent manner. Moreover, exogenous H2S rescued LPS-induced cystathionine gamma-lyase (CSE) inhibition and cystathionine beta-synthase (CBS) synthesis. Interestingly, in cells undergoing inflammation-induced apoptosis, H2S activated the PI3K/Akt and NF kappa B signal pathways both tested concentrations. Akt appeared to be a key crosstalk molecule that played a "bridge" role. Conclusions H2S regulates LPS-induced inflammation and apoptosis by activating the PI3K/Akt/NF kappa B signaling pathway. Hence, NaHS may be clinically useful for preventing or treating mastitis.