Study On Protective Effect And Mechanism Of Gastrodin On PC12Cell Injury Induced By Glutamate

Aim: To investigate the protective effect and mechanism of gastrodin on PC12cellsinjury induced by glutamate.Methods：PC12cells were treated with glutamate to establish injury model, MTT assay was usedto analyze cell viability to establish the best suitable concentration of the model.Inverted microscope was used to observe cell morphology, MTT assay to detect cellviability. Cell apoptosis morphological changes were detected by AO/EB staining.Changes of the morphology of PC12cells were detected by Hoechst33258stainingunder fluorescence microscopy. Release of intracellular reactive oxygen species (ROS)and changes of cell apoptosis rate via Annexin V/PI staining were analyzed by the flowcytometry. Changes of cell mitochondrial transmembrane potential were detected byRhodamine123staining. Western blotting to analyze expression of p-p38, p-ERK1/2and caspase-3protein.Results:1. Results of MTT assay indicated that growth inhibition rate of PC12cells nearly50%after treated with15mmol/L glutamate for24h.2. Results of MTT assay showed that gastrodin with different concentrations on injuredPC12cells induced by glutamate for72h showed a certain degree of protection,compared with the model group；The morphology observation results showed themorphology of gastrodin treated cells was closer to the normal cells.3. AO/EB staining showed that cells of control group showed a regular morphology,most cells were stained blue, the model group cells appeared orange fluorescence, cellsof gastrodin treated group showed less orange fluorescence, compared with the modelgroup.4. Hoechst33258staining showed that cells of model group were stained to bright bluefragment, which was a typical feature of apoptosis of cell nucleus, gastrodin treated group presented less of morphology of apoptosis, compared with the model group.5. Detection of ROS level and Annexin V/PI staining by the flow cytometry indicatedthat gastrodin can apparently reduce ROS level, drop cell apoptosis induced byglutamate. Rhodamine123staining results showed that gastrodin can enhance the levelof mitochondrial membrane potential to maintained the normal functions ofmitochondrial.6. Western blotting results indicated that, comparing with control group, gastrodintreated group can significantly down-regulate expression of p-p38, p-ERK1/2, and canalso drop the expression of caspase-3protein induced by glutamate.Conclusion:Gastrodin shows an apparent protective effect on apoptosis of PC12cells injury inducedby glutamate within a certain dosage range, its protective effect and mechanism may berelated to gastrodin can reduce the releasing of ROS to prevent the occurrence ofoxidative stress, protect the normal structure of cell membrane, stabilize the level ofmitochondrial membrane potential, reduce expression of p-p38, p-ERK1/2, anddown-regulated expression of caspase-3protein.