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作 者:邱平明[1] 王慧君[1] 刘超[1] 李学锋[1] 谭晓辉[1]
机构地区:[1]南方医科大学法医学研究所,广东广州510515
出 处:《南方医科大学学报》2007年第4期427-431,共5页Journal of Southern Medical University
基 金:国家自然科学基金(30572090);广东省科技攻关项目(2004B33801008)~~
摘 要:目的建立和比较海洛因成瘾与正常大鼠前额叶皮质(PFC)蛋白质双向电泳图谱,寻找和鉴定海洛因成瘾大鼠PFC中的差异蛋白表达谱。方法以固相pH梯度等电聚焦为第一向和垂直SDS-PAGE为第二向,分别对正常对照大鼠和海洛因成瘾大鼠的PFC蛋白质样品进行二维分离,2-D图谱经ImageMaster 2D 5.0软件分析,选取7个差异蛋白点用基质辅助激光解吸电离飞行时间质谱/串联质谱进行鉴定。结果经肽指纹图谱和串联质谱分析,并在NCBInr数据库检索,选取的7个蛋白点被成功鉴定为5种蛋白:成瘾组特有的是葡萄糖调节蛋白58和26s蛋白酶复合体亚基p40.5;成瘾组含量下调的蛋白是ATP合酶D链;Ndufa10和α-烯醇化酶在两组中发生相对分子质量和(或)等电点迁移。结论双向电泳结合质谱分析初步鉴定了大鼠前额叶皮质中与海洛因成瘾和神经毒性相关的差异蛋白,为进一步深入研究其病理机制奠定了基础。Objective To obtain two-dimensional gel electrophoresis maps of the prefrontal cortex (PFC) proteins of normal and heroin-addicted rats for identifying the differentially expressed proteins in the addicted rats. Methods Rat models of heroin addiction were established, and the proteins in the PFC underwent two-dimensional gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertical SDS-PAGE as the second dimension. ImageMaster 2D 5.0 analysis software, matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) or MALDI-TOF-TOF-MS as well as NCBInr database searching were performed to separate and identify the proteome in the rat PFC. Results Seven protein spots to represent 5 proteins were identified by PMF and MALDI-TOF-TOF-MS. Among those proteins, glucose-regulated protein (58 kD) and 26 S proteasome subunit p40.5 existed only in heroin-addicted rats, in which ATP synthase D chain was down-regulated. Ndufa10 and Enol were present in both groups, but their molecular mass and pI were different. Conclusion Immobilized pH gradient two-dimensional gel electrophoresis allows good reproducibility in separation and identification of the proteome in the PDF of heroin-addicted rats, which facilitates further investigation of pathogenic mechanisms of heroin addiction and central nerve injury.
分 类 号:R749.64[医药卫生—神经病学与精神病学]
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