leaf extract protects against H₂O₂-induced oxidative stress by increasing autophagy in PC12 cells

The effect of Apocynum venetum leaf extract (AVLE) on the nervous system has been widely studied, but its effect on injured neurons is not fully understood. In the present study, the protective effect of AVLE on injured neurons was determined. H₂O₂ was used to induce oxidative stress in PC12 cells and cell viability assays were used to determine the optimum concentration range of AVLE and its protective effects against oxidative stress. A live-dead assay was performed to confirm the effects of AVLE on oxidative stress. Subsequently, expression of apoptotic proteins including Bax and cleaved-caspase-3 were evaluated to determine whether AVLE affected apoptosis, and reactive oxygen species (ROS) levels were detected to determine the role of AVLE in H₂O₂ exposure. Furthermore, expression of autophagic proteins including LC3-II and p62 were detected to evaluate the effects of AVLE on autophagic activity, and cells were treated with 3-methyladenine (3-MA), an autophagic inhibitor, to identify the underlying protective mechanism of AVLE. The results showed that the optimum conditions to induce oxidative stress were treatment with 40 µM H₂O₂ for 2 h, and the suitable range of AVLE concentrations was shown to be 1-100 µg/ml. AVLE improved cell viability in PC12 cells following treatment with H₂O₂. AVLE reduced the expression of Bax and cleaved-caspase-3, and decreased ROS production. Furthermore, AVLE upregulated LC3-II expression and downregulated p62 expression, whereas treatment with 3-MA increased the levels of ROS and apoptotic proteins. These results suggest that AVLE may protect injured neurons against oxidative stress-induced apoptosis, and this effect may be associated with the reduction of ROS by increasing autophagy.