Effects Of The Volatile Oil Of Angelica Sinensis On Apoptosis And Autophagy Of Nerve Cell After Ischemia Reperfusion Injury

Objective:To evaluate the protective effect of the volatile oils of Angelica sinensis（VOAS）on the PC12 cells exposed to oxygen-glucose deprivation and reperfusion（OGD/R）by apoptosis and autophagy.Methods:1.The high differentiated PC12 Cells subjected to the hypoxia plus glucose deprivation followed by reoxygenation plus glucose were employed to investigate the protective effects of essential oils from Angelica sinensis.MTT assay was applied to assess cell viability,the activity of superoxide dismutase（SOD）and lactate dehydrogenase（LDH）,contents of malondialdehyde（MDA）were measured by kit.2.Flow cytometry was used to detect the apoptosis rate.The morphological apoptotic and autophagy characteristics of PC12 cells were observed by acridine orange/propidium iodide（AO/PI）double staining and AO（acridine orange）staining respectively.Autophagy and apoptosis-related proteins,as well as ERK_1/2 signal transduction pathway and PI3K/Akt/mTOR signal transduction pathway protein expression were measured by Western-blotting.Results:1.The results demonstrated that VOAS increased cell viability,decreased LDH activity and MDA content,whereas increased the activity of SOD in OGD/R treated PC12cells.2.Flow cytometry analysis showed that the apoptosis rate of VOAS-treated cell was reduced.After AO/PI staining,it can be seen under fluorescent microscope that VOAS improve the apoptosis.At the same time,AO staining results showed that autophagy were improved compared with the model group.3.Western-blotting analysis showed that VOAS up-regulated the expression of BCl-2,down-regulated the expression of Bax and Cleaved-Caspase-3,and the expression of LC3-II,Beclin-1 and Atg5 were decreased significangtly.4.Western-blotting analysis showed that the expression of related proteins of ERK_1/2signal transduction pathway and PI3K/Akt/mTOR signal transduction pathway were increased treated with VOAS.Conclusion:1.VOAS could improve the permeability of cell membrane and the ability of cells to resist hypoxia,reduce the lipid peroxidation of cells.2.VOAS could inhibit the apoptosis and autophagy levels after ischemia-reperfusion injury.3.VOAS protects PC12 cells from OGD/R-induced apoptosis and autophagy via activation of the ERK1/2 and PI3K/Akt/mTOR signaling pathway.