| Background: Alzheimer's disease (AD) is a neurodegenerative disease in central nervous system. AD is characterised by occult incidence and progressive deterioration of memory, so which is affect seriously on cognitive functions and the behavioral capacity of the aged. Chinese herbs for reinforcing spleen yin (Zi-Bu-Pi-Yin, ZBPY) decoction is produced on the fundamental of rescuing spleen yin, and our past research showed ZBPY decoction could prevent from aging and had significant protective effect on brain. Proteomics and the relative technology provide us a possible way to deeply realizing changes in the overall level of cell protein. Therefore the research of AD from the level of proteomics holds positive significance for further realizing the pathogenesis, screening the markers, early diagnosis, researching the mechanism of drug and so on.Objective: In order to correspond to disease with syndrome in clinic effectively, it is our aim to establish a rat model with both Alzheimer's disease and spleen deficiency syndrome, then to study the effect of ZBPY on it. Another aim is to realize the essence of AD with spleen yin deficiency syndrome and search effectual targets of ZBPY in the overall level of cell protein. So we applied the relative methods of proteomics to analyze the difference of protein expression of the hippocampus in AD with spleen yin deficiency syndrome group and treated by ZBPY group to get foundation of theory for investigating the molecular mechanisms of AD and spleen yin deficiency syndrome.Methods: Rats model of spleen Yin deficiency was set up by the improper diet, overstrain and consumption of body fluid. Then the incubated Aβ1-40 was injected into both hippocampus of each rat. The treated group was administered with ZBPY. Observe the changes of the rats at symptom in different phases. The behavioral abnormalities were investigated by step-down test and Morris water maze test. Qualitative changes of the protein expression in hippocampus were compared in normal control group, the AD with spleen yin deficiency syndrome model group and treated group by two-dimensional gel electrophoresis. Selected proteins were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).Results: 1. The two model groups of Spleen yin deficiency had the appearance of Yin deficiency leading to endogenous heat ,for example that increment in drinking water (P<0.01), objective elevation of the body temperature (P<0.01). The ability of learning and memory of the two AD groups was reduced conspicuously (P<0.05). 2. Totally 9 protein spots of hippocampus tissue in AD with spleen yin deficiency syndrome group were one and a half expressed than the normal group. Six protein spots showed higher expression and three spots showed lower expression. There are obvious differences in expression of two protein spots between the treated group and normal control group. One spot showed lower expression and the other was enhanced in abundance. The differential protein spots were identified by MALDI-TOF-MS and results showed the expression annexinⅢwas higher than control group, at the same time the expression of tubulin beta chain 15, dihydropyrimidinase-like 2 and guanine nucleotide-binding protein beta-1 subunit were lower than control group. After treated by ZBPY the differential expression spots of treated group were adjusted.Conclusion: 1. Try to perfect the methods to make the models of Alzheimer's disease and spleen deficiency syndrome, and then the changes of behavior and symptom were observed in syndrome, disease, and combined syndrome with disease. 2. The changes of some proteins expression level in rats'hippocampus may contribute to AD with spleen yin deficiency syndrome. AnnexinⅢ, tubulin beta chain 15, dihydropyrimidinase-like 2 and guanine nucleotide-binding protein beta-1subunit are possibly related to AD with spleen yin deficiency syndrome. 3. ZBPY improved the symptom of spleen yin deficiency and dysfunction of learning and memory in AD rat combined with Spleen yin deficiency. ZBPY has a good effect on it by regulating protein (cytoskeletal protein, signal transducer, energy metabolism associated protein, etc.) expression of hippocampus tissue of rats.
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