【作者】 杨明；

【导师】 王春友；

【作者基本信息】 华中科技大学 ， 外科学， 2006， 博士

【摘要】 目的利用蛋白质组学高通量的特点,研究胰腺增生的机制,以寻找糖尿病治疗研究的新靶点及筛选潜在胰腺干细胞标记物。方法通过大鼠胰腺大部分切除(Px)的方法建立胰腺增生模型,采用二维电泳(2-DE)分离比较Px术后第三天大鼠增生胰腺组织与假手术(Sx)非增生胰腺组织蛋白质的表达谱差异,用基质辅助激光解吸离子化飞行时间质谱(MALDI-TOF-MS)和肽质量指纹图谱(PMF)的方法对差异蛋白质进行鉴定。Western blotting分析蛋白质HSP47、Vimentin在PX大鼠胰腺组织中的表达,以验证2-DE的结果。结果2-DE显示SX和Px大鼠胰腺组织平均蛋白质点数分别为1369±28和1315±28个,其中共有91个蛋白质点出现1.5倍以上的表达差异。所有差异蛋白质点PMF鉴定出53种蛋白,其中Vimentin,cytokeratin 8(CK8),lymphocyte cytosolic protein 1(L-plastin),heterogeneous nuclear ribonucleoprotein A2/B 1(hnRNP A2/B 1)和L-arginine:glycine amidinotransferase(AGAT)与胚胎发育和细胞分化有关,这些蛋白质可能是胰腺干细胞潜在的候选标记物。结论蛋白质组学的方法利用其高通量的特点能有效发现潜在的胰腺干细胞生物学标记物。更多还原

【Abstract】 Objective A broad-range proteomic approach was applied to investigate the complexity of the mechanisms involved in pancreatic regeneration for identification of new targets of diabetes treatment and potential markers of pancreatic stem cells. Methods A regeneration pancreatic model was induced by 90% partial pancreatectomy (Px) in rats. Changes in the protein expression in rat regeneration pancreas at 3rd day after Px, as comparing to sham surgery (Sx), were analyzed by using two-dimensional gel electrophoresis (2-DE), mass spectrometry (MS), and peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching. HSP47 and Vimentin were analyzed in Px and Sx rat with Western blotting to confirm the results of 2-DE. Results The average spots in gels for Sx and Px pancreatic tissues were 1369±28 and 1315±28, respectively. Two-DE displayed 91 spots with at least 1.5-fold of differential expression at the time point of 3 days after pancreatectomy and 53 differentially expressed proteins were identified by PMF. Among them, Vimentin, cytokeratin 8 (CK8), lymphocyte cytosolic protein 1 (L-plastin), heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) and L-arginine: glycine amidinotransferase (AGAT) are associated with embryogenesis and cells differentiation and may be new potential pancreatic stem cells markers. Conclusions The proteome profiling technique provided a broad-based and effective approach for discovering potential biomarkers of pancreatic stem cells.更多还原