蛋白质指纹图谱技术探索硒诱导金属硫蛋白对小鼠肺纤维化的防护作用

Exploring early marker of pulmanary fibrosis and the protection of Se-MT by SELDI-TOF-MS

【目的】利用蛋白质指纹图谱技术探索硒诱导金属硫蛋白(metallothionein,MT)对小鼠肺纤维化的防护作用。【方法】利用表面增强的激光解析离子化-飞行时间-质谱(SELDI-TOF-MS)技术测定硒对MT的诱导,寻找小鼠肺纤维化的异常蛋白表达标志并评估硒代MT的防护作用。【结果】IMAC-Cu芯片表达的蛋白质中发现分子量为6277 Da的一组脱金属MT2蛋白表达增强证实硒对MT的高诱导作用;在WCX2和H4芯片中,发现肺纤维化小鼠中肺组织和血清中存在分子量分别为3880 Da和3 364 Da的两蛋白的表达与正常组比较明显受到抑制,硒诱导MT增高后上述异常表达均有改善,说明有机硒可能对肺纤维化诱导产生的蛋白异常具有一定的防护作用。【结论】利用蛋白质指纹图谱技术分析组织和血清的蛋白特异性表达可以作为肺纤维化疗效监测和评价的一类标志物,为今后进行更多的药物或方法筛选提供了一种可以尝试的实验模式。

【Objective】To explore the protein fingerprinting of metallothionein（MT）induced by selenium to protect the pulmanary fibrosis of mice.【Methods】To the effects induced MT by selenium in lung of mice were determined with surface enhanced laser desorption ionization-time of flight mass spectrometry（SELDI-TOF-MS） and the unnormal protein-labels of pulmanary fibrosis in tissue and blood serum of mice was found out and the preventive effects were evaluated by inducing Se-MT.【Results】Through SELDI-TOF-MS the protein fingerprinting of lung tissue on IMAC-Cu chip expressed that the peak current intensity in 6 277 Da was increased.In WCX2 and H4 chips,the ratio of peak current intensity in 3 880 Da and 3 364 Da was inhibited respectively in the injury group,but there were no significant changes of protein expression between Se treated group and control group.【Conclusions】The discovery of these marker proteins not only provides a novel powerful proteomic model,but also captures the early marker in forepart pulmonary fibrosis.This non-injury method could be widely used in both clinic trial and pre-clinic study in the future.