摘要
目的探讨五味子乙素对β淀粉样蛋白25-35(Aβ25-35)诱导褐家鼠肾上腺嗜铬瘤PC12细胞损伤的保护作用及可能机制。方法 20μmol.L-1Aβ25-35诱导PC12细胞损伤,加入5,10,25μmol.L-1五味子乙素,四甲基偶氮唑盐比色法(MTT法)检测细胞存活率,逆转录-聚合酶链式反应法(RT-PCR法)检测PC12细胞β淀粉样前体蛋白(APP)基因及空泡分选蛋白35(VPS35)基因在mRNA水平的表达,免疫细胞化学法测定APP及VPS35在蛋白水平的表达。结果 MTT结果显示,5,10,25μmol.L-1五味子乙素组PC12细胞存活率较Aβ25-35组高(P<0.05);RT-PCR、免疫细胞化学染色结果显示,Aβ25-35组较正常对照组APP、VPS35 mRNA和蛋白表达均上调(P<0.05);不同浓度五味子乙素组APP及VPS35 mRNA和蛋白表达较Aβ25-35组减少(P<0.05),VPS35与APP变化趋势一致。结论 5,10,25μmol.L-1五味子乙素可降低Aβ25-35对PC12细胞的损伤,该作用呈浓度依赖性,其机制可能与降低VPS35表达、减少sorLA含量、延长APP运输时间相关。
Objective To explore the protective effect of sehisandrin B on amyloid 13-protein25.35 （Aβ25-35） induced PC12 cell injury and probable mechanism. Methods PC12 cell injury was induced by 20 μmol·L-1 Aβ25-35 to establish a model of Alzheimer＇s disease. Then the cells were treated with 5,10 and 25 μmol·L-1 of schisandrin B. MTT assay,RT-PCR and immunoeytochemical staining were used to detect cell viability, expression of amyloid p-protein precursor （ APP ） and vacuolar protein sorting 35 （ VPS35 ） at mRNA level and protein level, respectively. Results MTT assay showed that the survival rate of PC12 cells in AI325_35 group increased dose-dependently with the treatment of schisandrin B at concentrations of 5, 10 and 25 μmol·L-1 （P〈0.05）. RT-PCR and immunocytochemical staining results showed that expression levels of APP and VPS35 at mRNA and protein levels were up-regulated in Aβ25-35 group compared with those in control group （ P〈0.05 ） , while the above indicators were decreased after the treatment with schisandrin B at different concentrations （P〈0.05）. APP and VPS35 showed a consistent trend of changes. Conclusion Schisandfin B of 5,10 and 25μmol·L-1 antagonizes the cellular injury induced by Aβ25-35 in a dose-dependent manner. This may be caused by decreased VPS35 expression,reduced sorLA content and extended APP transportation time.
出处
《医药导报》
CAS
北大核心
2013年第8期1010-1014,共5页
Herald of Medicine
