| 丘潔 高海青 劉師蓮
摘要 目的 研究哇巴因對血管內皮細胞（vascular endothelial cells,VEC）生長的不同作用，采用蛋白質組學方法闡述哇巴因誘導VEC雕亡作用的差异表達蛋白。方法 采用四甲基偶氮唑鹽(MTT) 法觀察哇巴因對VEC細胞生長的抑制作用; 應用流式細胞術觀察細胞雕亡; 比色法檢測Caspase3 活性。雙向凝膠電泳（2－DE）篩選差异蛋白，質譜鑒定差异蛋白。結果 哇巴因誘導VEC細胞雕亡是其抑制細胞生長的主要因素, 經蛋白質組學方法鑒定得出9個差异蛋白。結論 采用雙向電泳連接質譜的比較蛋白質組學方法可篩選哇巴因作用的靶作用蛋白,爲進一步研究其在心血管系統的病理生理機制提供新的分子理論基礎。
關鍵詞 哇巴因 比較蛋白質組學 細胞雕亡
Proteomics analysis of vascular endothelial cells apoptosis induced by ouabain
QIU Jie,GAO Haiqing,LIU Shilian
Abstract Objective To identify the differently expressed proteins involved in VEC apoptosis induced by ouabain using proteomics approaches.Methods MTT analysis was used to detect human umbilical vein endothelial cells (HUVEC) viability, caspase-3 activity analysis and flow cytometry were to confirm apoptosis induced by ouabain. Two-dimensional electrophoresis in conjunction with mass spectrometry was applied to revealed the differently expressed proteins in response to ouabain treatment. Results Apoptosis was primarily responsible for ouabain induced cell death. Proteomics analysis revealed 9 regulated proteins were accompanied by ouabain. Conclusions These data provide new target proteins related to ouabain actions and bring new sights into cellular and subcellular mechanisms of ouabain in cardiovascular disease.
Keywords ouabain; proteomics; apoptosis
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