联合蛋白质组学和代谢组学探讨兔脊髓缺血再灌注损伤的时间规律

Discuss on time regularity of ischemical reperfusion injury of rabbits′ spinal cord by proteomics combined with metabolism

目的:通过建立蛋白质组学和代谢组学的研究平台,研究和探讨脊髓缺血再灌注损伤分子机制。方法:建立兔脊髓缺血再灌注损伤模型,获取损伤和正常脊髓的蛋白质组双向电泳荧光图谱,观察差异蛋白的不同时间点的变化规律,建立缺血再灌注24 h与正常脊髓蛋白质组的匹配差异图谱,鉴定差异蛋白;根据蛋白组学得出的分析结果初筛,分别重点抽取假手术、术后30 min和术后24 h组不同时间段的血液制备血浆,采用气相色谱-飞行时间质谱仪(GC-TOF-MS)的方法进行分析,比较不同时间点的代谢水平变化。结果:蛋白质组学方法比较再灌注损伤24 h组与正常组可检测到46个蛋白差异点,鉴定出10个差异蛋白。代谢组学方法组检测出不同时间段兔血清中50种小分子物质的变化。再灌注损伤24 h组织代谢达到了高峰。结论:再灌注损伤24 h蛋白变化最明显,代谢达到峰值,是值得关注的关键时间点。

Objective： To discuss numerator mechanism of ischemical reperfusion injury of spinal cord by the method of proteomics combined with metabolism. Methods： Ischemical reperfusion injury model of rabbits＇ spinal cord were used to take proteomics two-dimensional electrophoresis photofluorogram of injured and normal spinal cord, then observed the change regularity of different time of different proteinum. Different spectrum of a graph of 24 h ischemical reperfusion in- jury and normal spinal cord proteome were raised to identify different proteinum. By the results, preparation blood plasma were adopted in sham operated team, 30 min after operation, 24 h after operation, then analyzed them by the method of gas chromatogram-time-of-flight mass spectrometer （GC-TOF-MS） to get the changes of metaboilic level in different time. Results： The method of proteomics could detect 46 proteinum disparate points compared with 24 h ischemical reperfusion injury and identify 10 different proteinum. The method of metabolism could detect the changes of 50 micromolecule sub- stance at different time, and architecture metabolism reached peak time at 24 h ischemical reperfusion injury. Conclusion： The changes of proteinum are the most obvious at 24 h ischemical reperfusion injury, and it is the most valuable to pay close attention to it.