西维来司钠对脑缺血/再灌注大鼠全脑蛋白质组的影响

目的:研究西维来司钠(Sivelestat sodium hydrate,ONO-5046)对脑缺血/再灌注(Cerebral ischemia/reperfusion,CI/R)大鼠全脑蛋白质表达的影响.方法:大鼠随机分为假手术组、CUR组和CI/R+ONO-5046组(6 mg/kg).采用四血管阻断法制备CUR模型,缺血15 min,再灌注24 h.提取脑组织总蛋白进行双向电泳,对差异蛋白点用HPLC-Chip-MS/MS纳流液质联用技术进行序列分析,经Spectrum Mill搜索NCBInr数据库后鉴定蛋白质.结果:成功获得了分辨率和重复性好的大鼠脑组织蛋白2-DE图谱,经质谱鉴定最终获得了巨噬细胞炎性蛋白1-beta、白细胞弹性蛋白酶A、β2-巨球蛋白、谷胱甘肽S-转移酶omega-1、谷氨酸脱氢酶-1、细胞色素C还原酶铁硫亚单位、硫氧还蛋白过氧化物酶1等19个差异蛋白点相关信息.结论:大鼠脑缺血后脑组织蛋白质表达存在明显差异,西维来司钠可通过调节有关蛋白质的表达,参与大鼠脑缺血神经元保护作用.

Effect of sivelestat sodium hydrate on proteomics in rats with global cerebral ischemia/reperfusion

Objective:To study the effect of Sivelestat sodium hydrate (ONO-5046) on brain proteomics in rats with cerebral ischemia/reperfusion(CI/R). Methods: SD rats were randomly divided into sham operated group, CI/R group and CI/R+ONO-5046 group(6 mg/kg).CI/R was induced by four-vessel occlusion method, with ischemia for 15 min, and reperfusion 24 h. After 2-dimensional electrophoresis and PDQuest for gel image analysis, analysis, the MS/MS of the peptides from the differential proteins digested by tripsin were analyzed by HPLC-Chip-MS/MS and the original data were pre-processed by Spectrum Mill then searched in NCBInr database. Results:Well resolved, reproducible 2-DE profiles of rat brain proteome were obtained. Among the differential protein spots, 19 were identified, Such as small inducible cytokine A4, Leukocyte elastase inhibitor A, Alpha-2-macroglobulin, Glutathione S-transferase omega-1, glutamate dehydrogenase-1 ,Thioredoxin peroxidase 1 ,Cytochrome b-cl complex subunit Rieske and so on. Conclusion:The cerebral proteomics expreesion after CI/R has obvious difference, Sivelestat sodium hydrate may promote the survival of injured neurons in rats by virtue of up or down-regulating the exprassion levels of proteins concerned with cerebral ischemia injury.