Abstract:Objective To investigate the effect of polydatin(PD) on oxidative stress-induced mitochondrial damage in alveolar epithelium and its possible mechanism. Methods A549 cells were assigned to four groups:control group in which cells were treated with 0.1% DMSO for 60min; model group in which cells received the pretreatment of 0.1% DMSO for 30min and exposure to H2O2 (250μmol/L) for 30min; treatment group in which cells received the pretreatment of PD (50μmol/L) for 30min and exposure to H2O2 (250μmol/L) for 30min; and inhibitor group in which cells received the pretreatment of PD (50μmol/L) and mitochondrial division inhibitor 1 (mdivi-1,10μmol/L) for 30min, and exposure to H2O2 (250μmol/L) for 30min. Western blotting was performed for expression of mitochondrial membrane protein [translocase of outer mitochondrial membrane 20 (TOM20), translocase of inner mitochondrial membrane 23 (TIM23)] and mitogen regulatory factor [mitochondrial transcription factor A (mTFA), and peroxisome proliferator-activated receptor-gamma coactivator-1α (PGC-1α)]. At the same time, Keima method was used to detect the red/green fluorescent area of cells as a reflection of mitochondrial autophagy. ATP was measured by fluorescein-luciferase kits, mitochondrial membrane potential (MMP) was determined using the fluorescent dye JC-1, reactive oxygen species (ROS) level was evaluated by DCFH-DA, and cell viability was determined by cell count kit-8 (CCK-8). SPSS statistics 20.0 was used for analysis. Results (1) Mitochondrial autophagy level. The expression of TOM20 and TIM23 was significantly lower in the model group than in the control group, in the treatment group than in model group, but significantly higher in the inhibitor group than in the treatment group (<0.01). However, there was no significant differences in mTFA and PGC-1 α between the four groups (P>0.05). Keima method showed that the change of red/green fluorescent area ratio in the four groups trended conversely with the changes of expression level of TOM20 and TIM23. (2) MMP. MMP was (100.0±5.9)% in the control group, (54.2±4.8)% in the model group, (70.8±3.6)% in the treatment group, and (56.0±6.1)% in the inhibitor group. Compared with control group, MMP in the model group decreased significantly, MMP in the treatment group increased significantly, while MMP in the inhibitor group decreased significantly (P<0.01). (3) ROS, cell activity and ATP level. Compared with control group, ROS in the model group increased significantly, while cell activity and ATP decreased significantly.Compared with the model group, ROS in the treatment group decreased significantly, while cell activity and ATP increased significantly. Compared with the treatment group,ROS level in the inhibitor group increased significantly, while cell activity and ATP decreased significantly (P<0.01). Conclusion PD can significantly improve oxidative stress-induced mitochondrial damage in alveolar epithelium, and its mechanism may be related to the up-regulation of mitochondrial autophagy.