Pterostilbene Inhibits ROS Production in Primary Spinal Cord Neurons by Activating Autophagy

Abstract: Autophagy is an important self-adaptive mechanism by inhibiting ROS (reactive oxygen species) in primary spinal cord neurons. Pterostilbene， a natural plant extract， has an antioxidant effect; however whether pterostilbene could protect spinal cord neurons from oxidative stress remains unclear. In the present study， primary spinal cord neurons of Sprague Dawley rats were seperated and cultured. CCK-8 analysis was used to detect the cytotoxicity. Spinal cord neurons were treated with pterostilbene in different doses for 24 h and 48 h， then LC3-II， Beclin-1 and P62 levels were determined by Western blot and the autophagy level was determined by transmission electron microscope. Primary spinal cord neurons were treated with H2O2 and pterostilbene for 24 h， then LC3-II level was determined by Western blot and the number of autophagosomes by GFP-LC3 analysis. DCFDA and MitoSOX Red staining were used to detect the ROS production in cells， and ATG5 siRNA transfection was used to analyze the involvement of autophagy. The results showed that there was a dose-dependent change in the levels of LC3-II， Beclin-1 and P62， and increased autophagosome number were observed in the pterostilbene-treated neurons (P<0.05). In addition， pterostilbene increased the level of LC3-II in cells treated with H2O2 (P<0.05)， and GFP-LC3 analysis demonstrated the increased number of autophagosomes in pterostilbene-treated cells. Compared with that in the cells treated with H2O2， pterostilbene significantly inhibited the ROS production; however， ATG5 siRNA transfection significantly reversed the protection of pterostilbene. These results indicate that pterostilbene inhibits the ROS production in spinal cord neurons by activating autophagy.

CSTR: 32200.14.cjcb.2017.08.0001