The Study On Protective Effects Of Biatractylolide On Glutamate-induced Neurotoxicity And Its Mechanism

OBJECTIVE: To study the protective effect of Biatractylolide on glutamate-induced human bone marrow neuroblastoma cell line(SH-SY5Y)and rat adrenal pheochromocytoma cell(PC12)injury,and to explore its mechanism,and provide a new theoretical basis for the further development of bialtosylactone as neurodegenerative diseases.METHODS: SH-SY5 Y and PC12 cells were cultured in vitro.The cells were treated with different concentrations of Glu for 24 hours.The model of SH-SY5 Y and PC12 cell injury and apoptosis induced by glutamate was established by MTT assay.The experiment was as follows:(1)blank control group(2)Glu injury model group(3)drug control group(low,medium and high).When cells were cultured to 70%,the cells were treated with serum-free for 12 hours.PC12 cells and SH-SY5 Y cells were incubated with 7.5 mmol/L glutamate and 10 mmol/L glutamate for 30 min before adding different concentrations of bacteol.The activity of lactate dehydrogenase(LDH)in culture supernatant was measured by spectrophotometry.The activity of ROS was detected by kit.Rhodamine123 was used to detect the change of mitochondrial potential.Annexin VApoptotic rate was detected by Annexin V-FITC / PI Apoptosis Detection Kit.Apoptotic morphology of cells was detected by acridine orange(AO)/ethidium bromide(EB)double staining fluorescent staining,and finally to detect the changes of Akt,P-Akt and GSK3?.RESULTS: When Glu(7.5mmol/L)was used to incubate PC12 cells for 24 h,the proliferation of SH-SY5 Y cells was significantly decreased and the growth of SH-SY5 Y cells was inhibited by Glu(10 mmol/L).After pretreatment with bilobal intraperitoneal lactone,the activity of cell proliferation was improved,the injury morphology was improved,the number of injured cells decreased,LDH release decreased,the apoptosis rate decreased,the ROS release activity decreased,the mitochondrial potential increased and concentration-dependent.Western blot analysis showed that the expression level of P-Akt protein and GSK3? protein were decreased.CONCLUSION:The Biatractylolide has a protective effect on glutamate-induced SH-SY5 Y cells and PC12 cells in a dose-dependent manner,and its mechanism may be through the inhibition of PI3k-Akt-GSK3? pathway.