Proteomics analysis of hippocampus and striatum in rats with hyperglycemia using iTRAQ technique

Q4 Medicine 中华内分泌代谢杂志 Pub Date : 2019-11-25 DOI:10.3760/CMA.J.ISSN.1000-6699.2019.11.010

null Maierhaba, Ablat Dilmurat, Xinling Yang, Y. Jia, Feifei Geng

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Abstract

Objective To screen the deferentially expressed proteins in hippocampus and striatum in rat models of diabetes mellitus and normal SD rats, and to elucidate the effects of hyperglycemia on central nervous system. Methods SD rats were randomly divided into normal group(n=10)and hyperglycemia group(n=15); rat models of diabetes were induced by the high-fat and high-sugar diet combined with single intraperitoneal injection of streptozotocin. The hippocampus and striatum tissues from hyperglycemia rat and normal SD rat(control group) were selected as specimens. Isobaric tags for relative and absolute quantification(iTRAQ) technique and 2-dimensional liquid chromatography-tandem mass spectrometry(2D-LC-MS/MS) were used for identifying the deferentially expressed proteins. The deferentially expressed proteins were analyzed by bio-informatics for searching candidate proteins.Finally, the candidate proteins were verified by Western blotting. Results A total of 347 proteins showed significantly different expressions, which included 139 up-regulated proteins and 289 down-regulated proteins. These proteins included metabolic glutamate receptor 2, ferritin, leucine repeat protein and complex 2, which are related to Parkinson′s disease. KEEN search indicated that the pathway of dopaminergic synaptic, complement, gap junction, autophagic and Fcgamma R-mediated phagocytosis were involved with these significant differentially expressed proteins. The protein interaction network showed that ferrin, aldehyde/ketone reductase(ARK), Ca2+ -binding protein(CaBP), and protein tyrosine phosphatase(PTP) were located at the crossed nodes. The expression levels of ferrin, ARK, PTP, and CaBP, which were verified by Western blotting, and the results were consistent with those of the quantitative mass spectrometry(P<0.05). Conclusion Deferentially expressed proteins in hippocampus striatum of diabetic rats can be effectively screened by iTRAQ technique combined with 2D-LC-MS/MS. It may provide new clues for the mechanism of neurodegenerative disease. Key words: Diabetes mellitus; Proteomics analysis; Isobaric tags for relative and absolute quantification; 2-dimensional liquid chromatography-tandem mass spectrometry

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应用iTRAQ技术对高血糖大鼠海马和纹状体的蛋白质组学分析

目的筛选糖尿病大鼠和正常SD大鼠海马和纹状体中不同表达的蛋白质，阐明高血糖对中枢神经系统的影响。方法SD大鼠随机分为正常组（n=10）和高血糖组（n=15）；采用高脂高糖饮食联合单次腹腔注射链脲佐菌素建立糖尿病大鼠模型。选择高血糖大鼠和正常SD大鼠（对照组）的海马和纹状体组织作为标本。采用相对和绝对定量同量标签（iTRAQ）技术和二维液相色谱-串联质谱法（2D-LC-MS/MS）对不同表达的蛋白质进行鉴定。通过生物信息学分析不同表达的蛋白质以寻找候选蛋白质。最后，通过蛋白质印迹法对候选蛋白质进行了验证。结果共有347个蛋白表达显著不同，其中139个蛋白表达上调，289个蛋白表达下调。这些蛋白质包括代谢型谷氨酸受体2、铁蛋白、亮氨酸重复蛋白和复合物2，它们与帕金森病有关。KEEN搜索表明，多巴胺能突触、补体、间隙连接、自噬和Fcgamma R介导的吞噬作用的途径与这些显著差异表达的蛋白质有关。蛋白质相互作用网络显示，铁蛋白、醛/酮还原酶（ARK）、Ca2+结合蛋白（CaBP）和蛋白质酪氨酸磷酸酶（PTP）位于交叉节点。结论应用iTRAQ技术结合2D-LC-MS/MS技术，可有效筛选糖尿病大鼠海马纹状体缺陷表达蛋白。它可能为神经退行性疾病的发病机制提供新的线索。关键词：糖尿病；蛋白质组学分析；用于相对和绝对定量的等压标签；二维液相色谱-串联质谱法

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来源期刊

中华内分泌代谢杂志 Medicine-Endocrinology, Diabetes and Metabolism

CiteScore

0.60

自引率

0.00%

发文量

7243

期刊介绍： The Chinese Journal of Endocrinology and Metabolism was founded in July 1985. It is a senior academic journal in the field of endocrinology and metabolism sponsored by the Chinese Medical Association. The journal aims to be the "Chinese broadcaster of new knowledge on endocrinology and metabolism worldwide". It reports leading scientific research results and clinical diagnosis and treatment experience in endocrinology and metabolism and related fields, as well as basic theoretical research that has a guiding role in endocrinology and metabolism clinics and is closely integrated with clinics. The journal is a core journal of Chinese science and technology (a statistical source journal of Chinese science and technology papers), and is included in Chinese and foreign statistical source journal databases such as the Chinese Science and Technology Papers and Citation Database, Chemical Abstracts, and Scopus.