应用蛋白质指纹图谱技术进行肝纤维化早期诊断的实验研究  

Experimental Study of Serum Proteomics Patterns Detected by SELDI-TOF MS in Early Diagnosis of Hepatic Fibrosis

作　　者：董永杰[1] 彭彦辉[2] 赵晓云 

机构地区：[1]邯郸市中心医院普外科,河北邯郸056001 [2]河北省人民医院普外科,石家庄050051 [3]河北省老年医学实验室,石家庄050051

出　　处：《中国普外基础与临床杂志》2010年第5期459-462,共4页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的检测肝纤维化各期的血清蛋白质组图谱,筛选特异的蛋白质标志物,用于肝纤维化的早期诊断。方法 SD大鼠50只,随机分为对照组(n=6)和实验组(n=44)2组,采用表面增强激光电离飞行时间质谱技术检测2组大鼠血清中的差异蛋白;采用放射免疫法检测血清肝纤维化4项,即Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)、层粘连蛋白(LN)和透明质酸酶(HA)。结果实验组中属肝纤维化Ⅰ、Ⅱ、Ⅲ及Ⅳ期者分别有8、5、5及6只〔有20只大鼠未到实验设定的采集标本时间死亡(系注射CCl4所致)〕。发现了质核比为4 203、4 658及7 400的3个差异蛋白标志物,其中质核比为4 658及7 400的2个蛋白质峰在肝纤维化Ⅰ期即明显升高,与对照组比较差异有统计学意义(P<0.05);而血清肝纤维化4项的水平在肝纤维化Ⅲ期或Ⅳ期才明显高于对照组(P<0.05)。结论蛋白质指纹图谱技术在肝纤维化的早期诊断以及标志物筛选方面有一定的价值,值得进一步深入研究。Objective To find a new specific marker that can be used to early diagnose hepatic fibrosis by detecting the change of serum protein in patients with hepatic fibrosis.Methods This research adopted 50 SD rats（25 males and 25 females）,and from which 6 rats were selected randomly（3 males and 3 females） as control group,last 44 rates were divided into four groups according to four pathological stages as hepatic fibrosis model group（experimental group）.Distinct proteins in serum were detected by surface-enhanced laser desorption/ionization-time of flight-mass spectra（SELDI-TOF-MS）.Radioimmunoassay was used to measure four parameters of hepatic fibrosis which were hyaluronidase（HA）,precollagen Ⅲ（PCⅢ）,laminin（LN） and collagen Ⅳ（Ⅳ-C）.Results Distinct proteins in serum were detected in 8 cases of stage Ⅰ of hapatic fibrosis,5 cases of stage Ⅱ,5 cases of stage Ⅲ,6 cases of stage Ⅳ,and 5 cases of control by SELDI-TOF-MS.Three protein peaks were found（M/Z： 4 203,4 658,and 7 400）.The peaks of M/Z 4 658 and 4 700 proteins were obviously increased in the stage Ⅰ of hepatic fibrosis（P〈0.05）,while the changes of hepatic fibrosis four parameters appeared in stage Ⅳ of hepatic fibrosis.Conclusion This method shows great potential for early diagnosing of hepatic fibrosis and finding better biomarkers to hepatic fibrosis.

关 键 词：肝纤维化 差异蛋白 蛋白质指纹图谱技术 蛋白质组学 蛋白质芯片 

分 类 号：R575.2[医药卫生—消化系统]