β-Asarone protects PC12 cells against OGD/R-induced injury via attenuating Beclin-1-dependent autophagy
β-Asarone protects PC12 cells against OGD/R-induced injury via attenuating Beclin-1-dependent autophagy
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DOI：
10.1038/aps.2012.35
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发表时间：
2012-06-01
影响因子：
8.2
通讯作者：
Zhang, Sheng
中科院分区：
文献类型：
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作者：
Mo, Zhen-tao;Fang, Yong-qi;Zhang, Sheng
目的：目的：探讨石菖蒲β-细辛醚对缺血性脑卒中模型PC 12细胞自噬的影响。在OGD前1 h开始给药，持续3 h。然后将细胞在无药物和全培养基中在常氧条件下孵育24 h。流式细胞仪检测Beclin-1、细胞内游离钙浓度（[Ca 2 +]（i））和线粒体膜电位（MMP）。使用MTT测定法测量细胞活力。结果：β-细辛醚（20、30、45 μ g/mL）或钙通道拮抗剂尼莫地平（10 μ mol/L）预处理可显著增加OGD/R处理的PC 12细胞活力和MMP，降低Beclin-1表达和[Ca ~（2+）]（i）。倒置相差显微镜下观察，β-细辛醚或尼莫地平预处理可明显增加细胞数量，改善细胞形态。结论：β-细辛醚对OGD/R诱导的PC 12细胞损伤具有保护作用，其机制可能与降低细胞内[Ca 2 +]（i）、增加细胞膜基质金属蛋白酶（MMP）而减轻Beclin-1依赖的自噬有关。
Aim: To explore the effects of beta-asarone from Acorus Tatarinowii Schott on autophagy in an ischemic stroke model of PC12 cells.Methods: The ischemic stroke model of PC12 cells was made by OGD/R (2 h oxygen-glucose deprivation followed by 24 h reperfusion). Drug administration was started 1 h before OGD and last for 3 h. Then the cells were incubated in the drug-free and full culture medium under normoxic conditions for 24 h. After the treatments, Beclin-1, intracellular free calcium concentration ([Ca2+](i)) and mitochondrial membrane potential (MMP) were analyzed using flow cytometry. Cell viability was measured using MTT assay. Cell morphology was studied under inverted phase contrast microscope, and autophagosomes were observed under transmission electron microscope.Results: Pretreatment with beta-asarone (20, 30, or 45 mu g/mL) or the calcium channel antagonist nimodipine (10 mu mol/L) significantly increased the cell viability and MMP, and decreased Beclin-1 expression and [Ca2+](i) in OGD/R-treated PC12 cells. Under inverted phase contrast microscope, pretreatment with beta-asarone or nimodipine dramatically increase the number of cells and improved the cellular morphology. Autophagosomes were found in OGD/R-treated PC12 cells as well as in drug plus OGD/R-treated PC12 cells.Conclusion: beta-Asarone protects PC12 cells against OGD/R-induced injury partly due to attenuating Beclin-1-dependent autophagy caused by decreasing [Ca2+](i) and increasing MMP.