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目的观察异丙酚对脂多糖(LPS)所致人单核细胞(THP1)损伤的蛋白质表达谱变化,探讨其可能的作用机制。方法将体外培养的THP1细胞分为对照组,1μg/mlLPS刺激组(L组),1μg/mlLPS加50μmol/L异丙酚组(L+P组),双向电泳分离三组THP1细胞中全部蛋白质,选择差异蛋白质点进行质谱分析,用Western blot对部分蛋白进行验证。结果经双向凝胶电泳和图像分析,发现9个蛋白点的表达水平在三组间有显著差异,经质谱分析得到4个蛋白质点的鉴定,即Stathmin 1、泛素-蛋白连接酶(ubiquitin-conjugating enzyme E2N)、(ChainA,I113t Mutant Of Human Sod1)和程序性细胞死亡因子6(programmed cell death6)。结论获得重复性和分辨率较好的THP1细胞双向凝胶电泳图谱,初步鉴定了部分与异丙酚保护LPS所致THP1细胞损伤作用相关的蛋白,为进一步了解异丙酚作用机制提供了新线索。
Objective To observe the changes of protein expression profile of propofol on lipopolysaccharide (LPS) -induced human monocytic cells (THP1) injury and to explore its possible mechanism. Methods THP1 cells cultured in vitro were divided into control group, LPS stimulation group (1μg / ml), LPS plus 1μg / ml group and LPS group (50μmol / L propofol group) , Different protein spots were selected for mass spectrometry analysis, and some proteins were verified by Western blot. Results Two-dimensional gel electrophoresis and image analysis showed that there were significant differences among the three protein spots among the three groups. Four protein spots were identified by mass spectrometry, namely Stathmin 1, ubiquitin- conjugating enzyme E2N), (ChainA, I113t Mutant Of Human Sod1) and programmed cell death 6. Conclusions Two-dimensional gel electrophoresis of THP1 cells with good reproducibility and resolution was obtained, and some proteins related to protection of LPS-induced THP1 cells induced by propofol were preliminarily identified, providing new clues for further understanding of the mechanism of action of propofol .