脂多糖诱导急性肺损伤小鼠肺脏蛋白质电泳图谱分析

Two-dimensional gel electrophoresis protein analysis of lipopolysaccharide-induced acute lung injury in mice

目的蛋白质双向电泳技术观察气管滴注脂多糖诱导急性肺损伤后小鼠肺组织在蛋白质水平上的改变。方法小鼠气管内滴注脂多糖制备小鼠急性肺损伤模型,48 h后做小鼠肺CT、肺病理切片及测肺湿/干重比用于观察肺损伤程度;并提取总蛋白,进行双向电泳测定,对获取的蛋白图谱进行分析,寻找差异表达的蛋白质。结果实验组肺CT显示明显炎症表现;病理切片显示小鼠肺组织水肿充血改变明显,肺泡中有大量红细胞和中性粒细胞浸润,肺损伤评分和肺湿/干重比正常组增加;双向电泳结果显示实验组小鼠比正常组小鼠的肺组织总蛋白数增加,蛋白质点数为(100±2),其中有21个表达的蛋白差异点,通过蛋白质谱鉴定和生物信息检索结果显示蛋白质过氧化物酶6在实验组和正常组中有明显差异,其可能与急性肺损伤的发生相关。结论通过向小鼠气管内滴注脂多糖可以诱发急性肺损伤,蛋白质双向电泳结果显示实验组小鼠肺组织中蛋白质过氧化物酶6表达的增加,机制还待进一步研究。

Objective To observe the changes of lung proteins level with acute lung injury （ALI） by intratracheal instillation of lipopolysaccharide with two-dimensional polypropylene gel electrophoresis（2DE）. Methods The ALI model was induced by intratracheal instillation of lipopolysaccharide, after 24 hours, the mouse computed tomo- graphy, histopathological changes in lung tissue and lung wet/dry weight ratio were observed. The lung total protein extracts of the two groups were analyzed by 2DE, then analyzed the comparison of electrophoregram and found out the differential expression of protein spots. Results The experimental mouse computed tomography showed lung in- flammation ; histopathological changes in lung tissue had higher level of pulmonary edema, red cell and neutrophil in- filtration in the pulmonary alveoli;lung injury score and lung wet/dry weight ratio of experiment group were signifi- cantly higher than that of control group. Results of 2DE indicated the lung total proteins in experiment group were higher than the control group. There were 21 diferential expression proteins in the （100 ±2） protein points, perox- iredoxin 6 was identified with LC-MS/MS and bioinformatics database of which was diferential expression proteins, and the expression changing of this protein may be involved in the acute lung injury. Conclusion ALI model could be established after intratracheal instillation of lipopolysaccharide,the results of 2DE show that the peroxiredoxin 6 is found upward-expressed in the experiment group, while the mechanism is still needed to be well studied.