当归挥发油通过抑制自噬减轻拟缺血再灌注神经细胞损伤

Protection of essential oils from Angelica sinensis on the neural cells of damaged by ischemia-reperfusion like injury through inhibition of autophagy

目的:探讨自噬在拟缺血再灌注损伤神经细胞中的作用以及当归挥发油对自噬的影响。方法:采用"缺糖-复糖"结合"缺氧-复氧"的方法,在体外用连二亚硫酸钠(Na2S2O4)处理高分化PC12细胞,模拟"缺血再灌注"损伤,用CCK-8比色法检测当归挥发油对细胞增殖的影响,AO荧光染色观察自噬细胞形态,Western-blot检测自噬相关蛋白Beclin-1、Atg5、LC3B-Ⅱ的表达。结果:在6.25~25μg/ml的浓度范围内,当归挥发能够显著提高PC12细胞存活率,减少受损细胞酸性自噬泡的数量,25μg/ml的当归挥发油能够抑制自噬相关蛋白Beclin-1、Atg5、LC3B-Ⅱ的表达。结论:当归挥发油可能通过抑制自噬减轻拟缺血再灌注神经细胞的损伤。

Objective： To investigate the role of autophagy in the neural cells of damaged by ischemia-reperfusion like injury and the effects of essential oils from Angelica sinensis on autophagy. Methods： 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. CCK-8assay was applied to assess cell viability,The morphological autophagy characteristics of PC12 cells were observed by AO（ acridine orange）staining,The expression of Beclin-1,Atg5 and LC3-Ⅱ proteins were measured by Western-blot. Results： The essential oils from Angelica sinensis treatment with 6. 25-25 μg/ml improved the survival rate of PC12 cells,reduced the number of autophagic vacuoles,and significantly down-regulated Beclin-1、Atg5 and LC3-Ⅱproteins. Conclusion： These findings suggested that the essential oils from Angelica sinensis prevents the ischemia-reperfusion like injury on the PC12 cells by inhibiting autophagy.