Watanabe Hideki, Hamada Hiromi, Yamada Naoki, Sohda Satoshi, Yamakawa-Kobayashi Kimiko, Yoshikawa Hiroyuki, Arinami Tadao
Department of Obstetrics and Gynecology, Institute of Clinical Medicine, University of Tsukuba, Ibaraki, Japan.
Proteomics. 2004 Feb;4(2):537-43. doi: 10.1002/pmic.200300565.
Preeclampsia is a pregnancy-specific syndrome and a major cause of maternal mortality. The pathophysiology of preeclampsia is unknown, and no proteome analysis of preeclampsia has been reported. We sought to identify proteins associated with preeclampsia using a proteomic technique and performed two-dimensional electrophoresis (2-DE) on sera from six patients with preeclampsia and six normal pregnant women, followed by comparison of the SYPRO Ruby-stained 2-DE profiles. A group of overexpressed spots was identified in the limited study set. Overexpressed spots were identified as clusterin by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) followed by peptide mass fingerprinting, a protein database search, and Western blot analysis. Additionally, sera of 80 preeclamptic women and 80 normal pregnant women were processed by immunoassay methods to confirm changes in clusterin concentrations quantitatively. Immunoassays showed that clusterin levels in the 80 preeclamptic women were significantly higher than those in the 80 controls (mean +/- SD; 1.62 +/- 0.46 times reference level in preeclamptic women vs. 1.30 +/- 0.46 times reference level in controls, P < 0.001). Proteomic analysis of serum proteins is a promising tool for studying preeclampsia pathophysiology and identifying proteins associated with preeclampsia.
子痫前期是一种妊娠期特有的综合征，也是孕产妇死亡的主要原因。子痫前期的病理生理学尚不清楚，且尚未见有关子痫前期蛋白质组分析的报道。我们试图使用蛋白质组学技术鉴定与子痫前期相关的蛋白质，并对6例子痫前期患者和6例正常孕妇的血清进行二维电泳（2-DE），随后比较SYPRO Ruby染色的2-DE图谱。在这个有限的研究组中鉴定出一组过表达的斑点。通过基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS），随后进行肽质量指纹图谱分析、蛋白质数据库搜索和蛋白质印迹分析，将过表达的斑点鉴定为聚集素。此外，采用免疫分析方法对80例子痫前期妇女和80例正常孕妇的血清进行处理，以定量确认聚集素浓度的变化。免疫分析显示，80例子痫前期妇女的聚集素水平显著高于80例对照组（均值±标准差；子痫前期妇女为参考水平的1.62±0.46倍，而对照组为参考水平的1.30±0.46倍，P<0.001）。血清蛋白质的蛋白质组学分析是研究子痫前期病理生理学和鉴定与子痫前期相关蛋白质的一种有前景的工具。
