应用表面增强激光解吸附离子化飞行时间质谱技术筛检博莱霉素肺纤维化大鼠血清蛋白标志物

目的鉴定博莱霉素肺纤维化大鼠模型血清差异表达蛋白,寻找肺纤维化疾病早期诊断标志物。方法48只雄性大鼠随机分为博莱霉素组和对照组,每组24只,分别于建模后7、14、28 d各取8只动物处死,取肺组织制作石蜡切片,苏木精-伊红和天狼星红染色,普通光学显微镜和偏振光显微镜观察肺纤维化反应;表面增强激光解吸附离子化飞行时间质谱和弱阳离子蛋白芯片检测血清样本,生物信息学软件分析获得的蛋白指纹图谱,鉴定差异蛋白表达。结果在博莱霉素组大鼠血清中找到4个差异表达的蛋白质,用生物信息学分析软件产生1个肺纤维化动物诊断模型,由质荷比分别为3 530.7、7 043.5、8 332.6和9 068.0的蛋白质组成,与对照组比较,其峰强度均显著升高。结论博莱霉素诱导的肺纤维化血清蛋白表达谱发现的4个差异表达蛋白有可能成为早期诊断博莱霉素肺纤维化大鼠模型的特异血清生物标志物。

Detection of serum proteomic biomarkers by surface enhanced laser desorption and ionization time-of-flight mass spectrometry in rats with pulmonary fibrosis induced by bleomycin

Objective To identify the differentially expressed proteins in the serum of rats with pulmonary fibrosis induced by bleomycetin,in order to look for the biomarkers for early diagnosis of fibrotic lung diseases.Methods Forty-eight male rats were randomly divided into bleomycin group and control group,twenty-four rats in each group.Eight rats were sacrificed at 7,14 and 28 days after modeling in each group,Paraffin-embedded lung sections were stained with hematine-eosin and Sirius red,pulmonary fibrosis were evaluated with common light microscopy and micropolarimete.The serum samples were detected by surface enhanced laser desorption and ionization time-of-flight mass spectrometry and weak cationic protein chip,bioinformatic soft was used to analyze and identify the differentially expressed proteins.Results Four differentially expressed proteins were detected in the serum of rats in bleomycin group,and one diagnostic models for bleomycin-induced pulmonary fibrosis were generated using software,consisting of four proteins with mass/charge ratio of 3 530.7,7 043.5,8 332.6 and 9 068.0,respectively,which all increased significantly in the serum of bleomycin-treated rats.Conclusion Bleomycin-induced pulmonary fibrosis show four differentially expressed proteins,the proteins may be the specific serum biomarkers of early diagnosis of rats with pulmonary fibrosis induced by bleomycin.