鲍曼不动杆菌脓毒症大鼠中性粒细胞的蛋白质组学研究

Proteomic analysis of neutrophils of rats with Acinetobacter baumannii sepsis

目的通过分离并鉴定鲍曼不动杆菌脓毒症大鼠中性粒细胞的差异表达蛋白质，以发现可能的在鲍曼不动杆菌脓毒症中起关键作用的蛋白和可用于早期诊断的鲍曼不动杆菌脓毒症标志物。方法提取正常大鼠中性粒细胞和鲍曼不动杆菌脓毒症大鼠中性粒细胞的总蛋白质，用双向凝胶电泳分离蛋白并进行比较。选择在鲍曼不动杆菌脓毒症中明显差异表达的蛋白点，行质谱分析，并选取蛋白加以免疫印迹验证。结果获得了分辨率和重复性均很好的凝胶蛋白图谱。筛选出的在鲍曼不动杆菌脓毒症中明显差异表达的50个蛋白点，共有41个蛋白点被成功鉴定，其中在鲍曼不动杆菌脓毒症中高表达的为24个，低表达的为17个。Western blot蛋白验证基本与质谱鉴定一致。结论鲍曼不动杆菌脓毒症大鼠中性粒细胞相对于正常大鼠中性粒细胞蛋白存在明显的差异，通过蛋白质组学方法筛选并鉴定出的这些蛋白质可能会成为用于鲍曼不动杆菌脓毒症早期诊断和治疗的分子靶点。

Objective To detect potential proteins which may play a key role in pathogenesis and early diagnosis for Acinetobacter baumannii sepsis, and compare the protein profile of neutrophils of nai＇ve rats with that of rats infected with Acinetobacter baumannii. Methods Using two-dimensional electrophoresis for separation of proteins, we compared the differences of the profile of total proteins from neutrophils of naive rats with that of rats infected with Acinetobacter baumannii. Differences in the profiles of expressed proteins were observed, and some of the proteins were isolated and verified by line mass spectrometry analysis and Western blot. Results Fifty proteins from the neutrophils of rats with Acinetobacter baumannii sepsis were found to be different from those of naive rats. Forty-one of these proteins were able to be identified successfully using two-dimensional electrophoresis, in which 24 were upregulated, and 17 were downregulated eompared with the proteins isolated from neutrophils of naive rats. Conclusion The protein profile of neutrophils in rats with Acinetobacter baumannii sepsis is significantly different from that of naive rats. These proteins can be identified by proteomie screening, and may be used as potential markers for early diagnosis and molecular targets for effective treatment of Acirtetobacter baurnannii sepsis.