红芪多糖对老年性痴呆细胞模型损伤保护作用的差异蛋白质研究

Study on the protective effect of Hedysari Radix Polysaccharides and the difference of proteins expression after the treatment in Alzheimer's disease cell model

目的:研究红芪多糖对Aβ1-42诱导的AD细胞模型保护作用及差异蛋白质的影响。方法:采用Aβ1-42建立AD细胞模型,HRP作用48h后,收集细胞,提取细胞总蛋白。采用双向电泳方法建立SH-SY5Y细胞的蛋白表达谱,银染后扫描凝胶获得蛋白图谱。使用PD Quest软件分析找出模型组和HRP处理组间的差异蛋白点,计算差异蛋白的等电点和理论分子量,对差异明显的蛋白点进行质谱鉴定。采用生物信息学方法对鉴定成功的蛋白质进行蛋白-蛋白相互作用网络分析。结果:建立了SH-SY5Y神经细胞的双向电泳蛋白表达谱。差异分析结果显示,模型组与HRP处理组间有22个差异蛋白点。应用质谱方法成功鉴定了4个差异蛋白质,分别为胆碱激酶a、Rag调节蛋白复合体LAMTOR1、转录因子蛋白AP-2-δ和铁蛋白,且Rag调节蛋白复合体LAMTOR1与铁蛋白存在相互作用关系。结论:红芪多糖对AD细胞模型的保护作用与其能够调节胆碱激酶a、Rag调节蛋白复合体LAMTOR1、转录因子蛋白AP-2-delta和铁蛋白的表达密切相关。

Objective： To study the protective effect of Hedysari Radix Polysaccharides（HRP） and the difference of proteins expression after the treatment in Aβ1-42-induced AD Alzheimer＇s disease cell model. Methods： AD cell models were induced using 5 μM Aβ1-42. Cells were harvested and total cellular proteins were extracted after 48 hours＇ HRP treatment. The protein expression profile of SH-SY5 Y cells was established by two-dimensional electrophoresis,and protein maps were obtained after silver staining and gel scanning. PD Quest software was used to analyze the different proteins expression between the drug group and the model group. The isoelectric point and theoretical molecular weight of the differentially expressed protein were calculated,the mass spectrometry was performed for the distinct protein points. Results： A two-dimensional electrophoresis protein profile of SH-SY5 Y neurons was established. There were 22 differential protein spots（P〈0. 05） between model group and HRP group. Four differential proteins（Choline kinase alpha,Ragulator complex protein LAMTOR1,Ragulator complex protein LAMTOR1,Ferritin） were successfully identified by mass spectrometry,and the Rag regulatory protein complex LAMTOR1 interacted with ferritin. Conclusion： Hedysari Polysaccharides has the protective effect on AD Cell Model,and the mechanism is closely related to the regulation of choline kinase a,Rag regulatory protein complex LAMTOR1,transcription factor protein AP-2-delta and ferritin.