Objective: To investigate the neuroprotective effect of cyanidin-3-Oglucoside( Cy-3G).
Methods: Hippocampal neurons were isolated from newborn rat and divided into different groups. ( 1) To estimate the neuroprotective effects, the cells were treated with various concentrations of Cy-3G( 10, 20, 40, 60, 80 and 100 μmol/L) prior to( 3 h) amyloid β protein( Aβ) 25-35 treatment. Neuronal viability in terms of mitochondrial activity was evaluated with the colorimetric tetrazolium salt( MTT) assay. ( 2) To explore the related mechanism, the cells were divided into normal control group, Aβ25-35 group, Aβ25-35 + Cy-3G co-incubation group and Cy-3G group, then the cell viability was analyzed by Lactate dehydrogenase( LDH) kit. The mitochondrial membrane potential and the intracellular reactive oxygen species( ROS) were respectively detected by Rhodamine 123 and DCFH-DA probe and measured by fluorescence spectrophotometry.
Results: ( 1) Relatively, 40 μmol/L Cy-3G and 5 μmol/L Aβ25-35 co-incubation for 3 h showed the best protection effect. ( 2) Compared with the control group, the LDH activity in cell media increased significantly in Aβ25-35 group( P < 0. 05), while Cy-3G co-incubation was able to reduce LDH activity in cell media significantly( P < 0. 05). Compared with the control group, the mitochondrial membrane potential of the Aβ25-35 group decreased significantly( P < 0. 05), Cy-3 G co-incubation could significantly inhibit the decrease of mitochondrial membrane potential of hippocampal neurons induced by Aβ25-35( P < 0. 05). Compared with the control group, the levels of intracellular ROS in the Aβ25-35 group increased significantly( P < 0. 05), while Cy-3G co-incubation could significantly inhibit the increase of intracellular ROS levels( P < 0. 05).
Conclusion: The right amount of Cy-3G can protect the primary hippocampal neurons from injury induced by Aβ25-35.
Keywords: amyloid beta protein 25-35; cyanidin-3-O-glucoside; hippocampal neurons; reactive oxygen species.