The ethyl acetate fraction of Polytrichum commune L.ex Hedw induced cell apoptosis via reactive oxygen species in L1210 cells

Abstract

Ethnopharmacological relevancePolytrichum commune L.ex Hedw is a traditional Chinese herb for treatment of fever, hemostatic, uterine prolapse and especially for leukemia. Previous studies indicated its anti-leukemia effect but the potential mechanisms have not been fully explained. Aims of the studyThe present study was further to investigate the underlying mechanism of ethyl acetate extract of Polytrichum commune L.ex Hedw (EEF)-induced toxicity and apoptosis in L1210 cells. Materials and methodsViability, DNA damage and apoptotic protein expressions of L1210 cells were analyzed by ViaCount, comet assay and western blot, respectively. At different times after EEF treatment, Bax redistribution in L1210 cells was examined using confocal microscopy; loss of mitochondrial membrane potential (MMP) was monitored by fluorescence microscope using rhodamine 123 staining; Intracellular reactive oxygen species (ROS) generation and DNA fragmentation were measured by flow cytometry using fluorescent dye, DCFH-DA and PI, respectively. ResultsEEF significantly inhibited L1210 cell survival, promoted Bax translocation onto mitochondria, stimulated caspase-9 activation and subsequent DNA damage in L1210 cells. Abundant ROS was detected in L1210 cells after EEF treatment, and the ROS scavenger NAC significantly relieved EEF-induced cell viability decline, MMP loss, and DNA fragmentation. ConclusionEEF could induce mitochondria-dependent cell apoptosis in L1210 cells, and ROS may play an important role in this action.