Empirical Study of Proteome of Hippocampus from A Rat Model with Alzheimer's Disease with Spleen Deficiency Syndrome Effected by Zibu Piyin Recipe

Zhan Libin , Lu Xiaoguang , Liu Li , Gong Xiaoyang , Liang Lina , Shi Xiang
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Abstract

The article is aimed at investigating Alzheimer's disease (AD) with spleen yin deficiency syndrome and identifying the effective therapeutic targets of Zibu Piyin Recipe (ZBPYR) at whole cell protein levels. The rat model of spleen yin deficiency syndrome was set up through combination with improper diet, overstrain, and consumption of body fluid. Then, the polymerized β-amyloid 1–40 was injected into both hippocampi of the rats. The treated group was administered with ZBPYR. Taking the control group as a standard, the changes of proteins in the hippocampus were compared between the model group of AD with spleen yin deficiency syndrome and ZBPYR-treated group by two-dimensional gel electrophoresis. The altered protein spots were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Compared with the control group, 9 protein spots of the hippocampus in the model group of AD with spleen yin deficiency syndrome and 2 spots in ZBPYR-treated group were found to be changed totally. The differential protein spots were identified by MALDI-TOF-MS. The results showed that in the model group of AD with spleen yin deficiency syndrome, the level of annexin III was higher, but the levels of tubulin beta chain 15, dihydropyrimidinase-like 2, and guanine nucleotide-binding protein beta-1 subunit were lower than that of the control group. These differential proteins did not show significant changes between the ZBPYR-treated group and the control group. It is concluded that changes of several proteins in the hippocampus may involve in the pathogenesis of AD with spleen yin deficiency syndrome together. Annexin III, tubulin beta chain 15, dihydropyrimidinase-like 2, and guanine nucleotide-binding protein beta-1 subunit are possibly related to pathological changes of the hippocampus in AD with spleen yin deficiency syndrome. ZBPYR may produce its effects by regulating these proteins in the hippocampus.

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滋补脾虚方对阿尔茨海默病脾虚证大鼠模型海马蛋白质组的影响
本文旨在从全细胞蛋白水平探讨补补补阴方治疗阿尔茨海默病(AD)脾阴虚证的有效靶点。采用饮食不当、过度劳损、补液相结合的方法建立脾阴虚证大鼠模型。然后,将聚合的β-淀粉样蛋白1-40注射到大鼠的两侧海马。治疗组给予ZBPYR。以对照组为标准,采用双向凝胶电泳法比较AD合并脾阴虚证模型组与zbpyr治疗组海马组织蛋白的变化。用基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)分析改变后的蛋白斑点。与对照组相比,AD脾阴虚证模型组海马组织中有9个蛋白斑完全改变,zbpyr治疗组海马组织中有2个蛋白斑完全改变。用MALDI-TOF-MS鉴定差异蛋白斑点。结果显示,AD伴脾阴虚证模型组膜联蛋白III水平升高,而微管蛋白β链15、二氢嘧啶酶样2、鸟嘌呤核苷酸结合蛋白β -1亚基水平低于对照组。这些差异蛋白在zbpyr处理组和对照组之间没有显着变化。由此可见,海马内多种蛋白的改变可能共同参与了AD伴脾阴虚证的发病过程。膜联蛋白III、微管蛋白β链15、二氢嘧啶酶样2、鸟嘌呤核苷酸结合蛋白β -1亚基可能与AD伴脾阴虚证海马的病理改变有关。ZBPYR可能通过调节海马体中这些蛋白产生作用。
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