AIM: To investigate the differential phosphorylation and activation of p38 in hepatocytes by pro-apoptotic Transforming Growth Factor-β1 (TGF-β1), pro-survival factors Epidermal Growth Factor (EGF) and 12-O-tetradecanoylphorbol-13-acetate (TPA) and the potential mechanisms.METHODS: The phosphorylation and activation of p38were determined by immunoblotting. Apoptosis was analyzed by morphological staining and observation, FACS analysis of sub-G1 content and DNA fragmentation assay.To quantitatively determine caspase activation, caspase activity assay was performed in vitro.RESULTS: TGF-β1-induced apoptosis was associated with the phosphorylation of p38, and SB202190, a specific inhibitor of p38, which was able to inhibit TGF-β1-induced caspase activation and apoptosis. TPA and EGF also blocked apoptosis induced by TGF-β1. Both of them induced the phosphorylation of p38. The results showed SB202190 had no effect on TGF-β1-induced phosphorylation of p38, but effectively inhibited both EGF and TPA-induced phosphorylation of p38.CONCLUSION: Pro-apoptotic TGF-β1, anti-apoptotic TPA and EGF induce the phosphorylation of p38 through different mechanisms that can be distinguished by SB202190. The data suggest that TPA and EGF-induced p38 phosphorylation is through an autophosphorylationdependent mechanism. Since p38 phosphorylation induced by TGF-β1 plays an important role in caspase activation and apoptosis, TPA and EGF-induced p38 phosphorylation may not be requisite for their anti-apoptotic function.