The Effect Of Hedysari Radix Polysaccha- Ride On Protein Expression Profiling Of Alzheimers’ Disease Cells Model

Objective: To study whether the Hedysari radix polysaccharide (HRP) can improve the cells model of dementia (Alzheimer’s diseases, AD) viability and whether it can effect on protein expression profiling of Alzheimers’ disease cells mode.Methods: 1. To determine the toxic effects from the different concentrations of HRP acting on the SH-SY5Y neural cells by MTT assay.The SH-SY5Y neural cells were treated with doses of 5μM Aβ1-42, the treatment of different concentrations of HRP were determined by MTT assay and each group SH-SY5Y cells morphological changes were observed through inverted phase contrast microscope at the same time. 2. The SH-SY5Y cells proteins were purified using acetone precipitation method, then the proteins were separated through two-dimensional gel electropho-resis in optimized condition, then the protein profiles were obtained by silver staining and scan.3. Set in the control group,5μM Aβ1-42 model group and HRP drug intervention group, these proteins of SH-SY5Y cells were separated utilizing 2-DE in accordance with the experimental exploration of condition, analyzed the protein profiles through PD Quest 8.0 software carefully, discovered the different protein spots and identified further proteins by mass spectrometry.Results: 1. HRP nontoxic dose range is less than or equal to 80μg/ml; the AD cells models were successfully estabished by 2.5,5 or 10μM Aβ1-42 induced cause; the cells survival rate was increased significantly in 20, 40, 80μg/ml HRP drug groups than AD model group. 2. The SH-SY5Y cells proteins were purified using acetone precipitation method and separated in the optimal focusing condition, successfully established SH-SY5Y types of nerve cell protein two-dimensional electrophoresis conditions. 3. The outcomes of the 2-DE mappings with PD Quest 8.0 software are as follows: there were 912±23 protein spots expressed in the control group, 810±11 protein spots expressed in the model group, and 871±41 protein spots in HRP drug group, there were obvious differences 22 protein spots expression between the model group and HRP group, further there were four proteins which were identified through mass spectrome, they were Choline kinase alpha、Ragulator complex protein LAMTOR1、Transcription factor AP-2-delta and Ferritin.Conclusion: HRP can improve the cells model of dementia survival rate, its mechanism which may be related to impressed expression of choline kinase alpha, Ragulator complex protein LAMTOR1, Transcription factor AP-2-delta, Ferritin.