益智健脑颗粒水提浓缩液对P8系快速老化小鼠内嗅区组织差异蛋白质表达的影响

Effect of Yizhi Jiannao Granule Concentration Fluid on the Differential Expression Protein in Entorhinal Area Tissue of Senescence Accelerated Mouse P8

目的分析益智健脑颗粒水提浓缩液(Yizhi Jiannao Granule Concentration Fluid,YJGCF)干预前后P8系快速老化小鼠(senescence accelerated mouse P8,SAMP8)内嗅区组织的差异蛋白质表达,从蛋白质组学水平探讨中药益智健脑颗粒水提浓缩液治疗阿尔茨海默病(Alzheimer′s disease,AD)的作用机制。方法 6月龄SAMP8随机分为治疗组、模型组和对照组,每组10只。模型组不予处理,让其自由饮水,治疗组以YJGCF灌胃(0.3g/d),对照组以等剂量双蒸水灌胃。8周后、应用双向凝胶电泳技术分别分离3组SAMP8内嗅区组织总蛋白,得到差异蛋白质点,通过质谱分析得到相应肽质量指纹图谱,经蛋白质数据库查询,鉴定差异蛋白质点。结果治疗组与模型组存在32个差异点,初步鉴定了NADH脱氢酶铁硫蛋白6、Rho-GDP解离抑制因子α、β2珠蛋白、磷酸甘油酸激酶等14个差异蛋白质,其功能涉及线粒体能量代谢、氧化应激、神经元功能等方面。结论 YJGCF可调节SAMP8内嗅区组织的多种蛋白质表达,提示该药具有多靶点治疗的作用,且可能通过改善线粒体功能障碍、抗氧化应激、抑制神经细胞凋亡、保护神经元等途径来实现其治疗AD的作用。

Objective To explore the mechanism of Yizhi Jiannao Granule （YZJN） in treating Alzheimer＇s disease （AD） on proteomic level by analyzing the differential expression proteins in entorhinal cortex tissue of senescence accelerated mouse P8 （SAMP8） treated with YZJN. Methods Six-month old SAMP8 were randomly divided into 3 groups, the model group, the YZJN group and the control group, 10 mice in each group. The model group was untreated with free water access, the YZJN group was treated with concentrated water ex- tract of YZJN 0. 3 g per day via gastric perfusion, and the control group was perfused with equal volume of double distilled water. The total protein in entorhinal codex tissue of mice was extracted after an 8-week treatment with two-dimensiona3 gel electrophoresis, and the differential expression protein spots were separated for identification through peptide mass fingerprint analysis and database searching. Results Thirty-two protein spots expressed differentially between the YZJN group and the model group were found, and 14 differential expression proteins were identified, including NADH dehydrogenase iron-sulfur protein 6, Rho-GDP dissociation inhibitor alpha, beta- 2-globin, phosphoglyceric kinase, etc, their functions involved mitochondria energy metabolism, oxidative stress and neuron function. Conclusion YZJN could regulate multiple protein expressions in entorhinal codex tissues of SAMPS, suggesting that it has multi-target therapeutic action and its mechanism in treating AD is possibly real- ized by way of improving mitochondria function, antagonizing oxidation stress, preventing nerve cell apoptosis and protecting neurons.