脑出血早期病灶周边脑组织的差异表达蛋白

Differentially expressed proteins in periferal brain tissue after early cerebral hemorrhage

目的利用蛋白质组学技术对脑出血大鼠模型出血周围脑组织进行动态研究,旨在比较出血超急性期和急性期的脑组织差异表达蛋白。方法选择Wistar大鼠12只,利用立体定向仪制作大鼠尾状核脑出血模型,随机分为脑出血6 h组和脑出血72 h组,每组6只。取病灶周围脑组织为样本,提取脑组织蛋白,进行双向凝胶电泳及染色;胶内蛋白质酶解,所得肽段用基质辅助激光解析电离飞行时间质谱仪检测,得到的肽片段质量指纹图谱在蛋白质数据库检索,鉴定出差异表达蛋白。结果脑出血6 h组与脑出血72 h组41种蛋白质的表达量差异有统计学意义(P<0.05)。其中脑出血72 h组血肿周边脑组织有19种蛋白质表达较脑出血6 h组明显上调,主要包括磷酸甘油酸激酶1,延长因子Tu,抑制素,T-复合物蛋白1-ε等;在血肿周边脑组织有22种蛋白质表达较脑出血6 h组明显下调,主要包括热休克蛋白90β-1,电压依从性阴离子选择性通道蛋白1,肽基-脯氨酰顺-反异构酶等。结论本研究鉴定出不同时期的部分差异表达蛋白,包括可能参与脑出血后应激反应的蛋白以及炎性反应和凋亡相关的蛋白。有利于临床在蛋白质水平上对脑出血后疾病的病理损伤机制全面整体的认识。

Objective To compare the differentially expressed proteins in periferal brain tissue after early cerebral hemorrhage by establishing a rat cerebral hemorrhage model.Methods Twelve Wistar rats were randomly divided into 6 h cerebral hemorrhage group and 72 h cerebral hemorrhage group（6 ineach group）.A rat cerebral hemorrhage model was established.Periferal brain tissue samples were taken from hemorrhage foci with brain tissue protein extracted.The brain tissue samples underwent 2-dimensional gel electrophoresis after staining.The electrophoresis pattern was analyzed using the ImageMaster 2D Elite software.The proteins of interest were digested and detected by MALDI-TOF mass spectrometry.The differentially-expressed priteins were identified.Results The expression levels of 41 proteins were significantly different in 6 h cerebral hemorrhage group and 72 h cerebral hemorrhage group（P〈0.05）.The expression levels of 19proteins（such as phosphoglycerate kinase 1,elongation factor tu,prohibitin,t-complex protein 1ε）were significantly higher wheas those of 22 proteins（such as HSP90β-1,voltage-dependent anion channel,peptidyl-prolyl cis-trans isomerase A） were significantly lower in 72 h cerebral hemorrhage group than in 6 h cerebral hemorrhage group.Conclusion Certain differentially-expressed proteins we identified in this study include stress reaction protein,flammatory reaction protein and apoptosis-related protein,which can thus help us to have a comprehensive understanding of the pathological mechanisms of ICH at the protein level.