Fig. 1
L. japonica extract (LJE) decreases AA + iron induced cytotoxicity in HepG2 cells. HepG2 cells were incubated with indicated dose of LJE for 1 h and later treated with 10 μM AA for 12 h, being followed by exposure to 5 μM iron for 3 h. (a) Cell viability was assessed by the MTT assay. (b) Expression of proteins associated with apoptosis was determined by western blot analysis. Equal protein loading was verified by β-actin. (c) The levels of apoptosis in each groups examined by TUNEL assay. Representative images show apoptosis of HepG2 cells in vehicle control, LJE treated, AA + iron treated, and AA + iron treated with LJE groups (left). Percentage of TUNEL+ cell nuclei calculated relative to total number of cell nuclei (right). (d) Cellular GSH content was assessed in cells by using GSH assay kit. (e) Cellular reactive oxygen species production was monitored by measuring intensity of dichloro fluoresce in fluorescence. (f) ΔΨm depolarization monitored by FACS analysis of Rh123 staining. Relative proportions of low Rh-123 intensity (RN1 fractions) are expressed as the mean ± S.D. of three separated experiments. For panel from A to E, data represent the mean ± S.D. for the four replicates. ** p < 0.01