Objective: To establish a nerve cell injury model by incubating primary rat cortical neurons in the presence of hydrogen peroxide (H2O2) to study the effect of emodin on apoptosis of nerve cells.
Methods: Cortical neurons were incubated with H2O2 and emodin. The cell viability was measured by MTT and cell injury was evaluated by lactate dehydrogenate (LDH). Flow cytometry was used to evaluate the intracellular level of reactive oxygen species (ROS). The morphology of cortical cells were observed by inverted microscope and Hoechst 33258 staining. The expression of Bcl-2 and Bax were determined by western blot.
Results: The 150 micromol/L H2O2 significantly decreased the viability of cortical cells and increased the level of intracellular ROS compared to control. Hoechest 33258 fluorescent staining showed brightness nucleosomes in apoptosis cells. The injury state was ameliorated by pretreatment of emodin. Moreover,as detected by Western blot analysis, H2O2 down-regulated the expression of Bcl-2 and up-regulation the expression of Bax compared to control. The state could be reversed by pretreatment of emodin.
Conclusion: Emodin can resist the adverse effects of H2O2 on increasing apoptotic rates, and it has the protective function towards nerve cells.