Protective Effect of Negletein Against Hypoxia Induced Injury on PC12 Cells

OBJECTIVE to investigate the protective effect of negletein against hypoxia induced injury on PC12 cells. METHODS The PC12 cells were divided into four groups, including normal control group, hypoxia model group, rutin group and negletein group. Cell viability was detected using MTT assay. Lactate dehydrogenase (LDH), malondialdehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) in cell culture supernatant were evaluated by enzyme labeling method. Reactive oxygen species (ROS) level was evaluated by DCFH-DA. The expression of mRNA and protein of Nrf2 and heme oxygenase-1(HO-1) were assessed with RT-PCR analysis and Western blot. RESULTS Compared with normal control group, the viability as well as the activity of antioxidant enzyme such as SOD and CAT were reduced and the level of LDH, MDA and ROS were increased treated with hypoxia in PC12 cells. These changes were reversed with the pretreatment of negletein. Moreover, the expression of Nrf2 and HO-1 mRNA and proteins were also increased induced by hypoxia, even further upregulated by negletein pretreatment. CONCLUSION The results indicate that negletein can suppress the hypoxia-induced damage on PC12 cells by activating of the Nrf2/HO-1 pathway and alleviation of oxidative stress.