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目的对比四种开窍药的有效成分对β-淀粉样蛋白42(Aβ_(1-42))诱导PC12细胞损伤的细胞内Ca~(2+)水平、线粒体膜电位(MMP)以及自噬相关蛋白Beclin-1的影响。方法采用Aβ_(1-42)诱导PC12细胞制备体外模型,以四种醒脑开窍药的有效挥发性成分β-细辛醚、冰片、麝香酮、苏合香挥发油作用24 h,光镜观察细胞形态,CCK-8法检测细胞活力,流式细胞仪检测细胞内Ca~(2+)水平、MMP和Beclin-1。结果造模后细胞形态不规则,数量减少,胞体略小,突起缩回,β-细辛醚、冰片、苏合香挥发油、麝香酮能改善损伤的细胞形态,提高细胞活力(P<0.05),降低细胞内Ca~(2+)水平及升高MMP(P<0.05),升高Beclin-1蛋白的含量(P<0.05)。结论四种开窍药的有效成分β-细辛醚、冰片、苏合香挥发油、麝香酮对Aβ1-42诱导的细胞模型有保护作用,可降低损伤细胞的游离Ca~(2+)水平、升高MMP、升高Beclin-1蛋白。
OBJECTIVE: To compare the effect of four kinds of resuscitation drugs active ingredients on intracellular Ca 2+, mitochondrial membrane potential (MMP) and autophagy-related protein (PC-2) induced by β-amyloid 42 (Aβ 1-42) Influence of Beclin-1. Methods PC12 cells were induced by Aβ 1-42 to prepare the in vitro model. The effects of β-asarone, borneol, musk ketone and volatile oil from four kinds of drugs on resuscitation were observed. Cell viability was detected by CCK-8 assay. Intracellular Ca 2+, MMP and Beclin-1 were detected by flow cytometry. Results The morphology of the cells was irregular, the quantity was reduced, the cell bodies were slightly smaller and the processes were retracted. The β-asarone, borneol, fulgurone and musk ketone could improve the cell morphology and the cell viability (P <0.05) (P <0.05), and increase the content of Beclin-1 protein (P <0.05). CONCLUSION The active ingredients of the four resuscitation drugs, β-asarone, borneol, fulguride and musk ketone have a protective effect on Aβ1-42-induced cell model, which can reduce the level of free Ca2 + in injured cells and increase the MMP , Increase Beclin-1 protein.