食蟹猴局灶性脑缺血模型中脑组织蛋白质组学鉴定分析研究

A Proteomic study of focal cerebral ischemia of brain tissues in macaque monkey

目的探讨食蟹猴局灶性脑缺血模型中梗死区、半影边缘区及正常组织之间蛋白质组学的差异。方法成功建立灵长类动物食蟹猴脑缺血模型4 h后,提取正常区、半影区及梗死区组织,采用强型阳离子交换法在蛋白水平进行分级,随后使用纳升级超高效液相色谱串联沃特式高分辨质谱进行数据采集。后期数据经过去冗余化和分类,并进行代谢通路分析。结果在缺血组织、半影区组织、正常组织中分别鉴定得到178、133、282个非冗余蛋白。通过KEGG通路分析,在3个区域中差异性表达的蛋白质参与最多的通路包括代谢通路、免疫反应通路和转录调节通路。参与物质代谢的蛋白在3个区域中的表达个数有明显的不同。梗死组织表达了更多种类的免疫相关蛋白,而在半影区组织中则表达了更多的转录调节因子。结论与梗死区相比,边缘区的组织损伤可逆,是使用药物治疗脑缺血的重点。脑缺血4 h后,正常组织、半影区、梗死区组织的蛋白质表达以及蛋白参与的代谢通路均具有明显差异。其结果为解释脑缺血病理机制提供了信息,为疾病治疗药物的开发提供了靶标分子。

Objective To explore the differences in proteomics among the normal,ischemic penumbra and ischemic infarction brain tissues of macaque monkey. Methods The normal,penumbra and ischemic tissue samples were collected from a macaque subjected to cerebral ischemia for 4 h. Proteins were extracted and fractionated using strong cation exchange chromatography. Proteins were digested and analyzed by nanoUPLC coupled to high definition mass spectrometry. Non-redundant proteins were grouped and pathway analysis was conducted using KEGG. Results Proteins were differentially expressed among different regions. More immunity-associated proteins were expressed in the ischemic infarction tissues,suggesting severe immunologic or inflammatory reactions in severe injured regions. On the contrary,more transcriptional factors were expressed in the penumbra region,which suggested that in the mildly injured region,gene transcription was more up-regulated to protect the tissue. Conclusion Compared to theischemic infarction region,injury in the penumbra region is reversible. After 4 h cerebral ischemia,the expression of proteins and metabolic pathways involved are different in different regions. These results provide a critical evidence on pathogenic mechanism of brain ischemia and potential targets for ischemia treatment.