摘要
目的比较胆道闭锁与淤胆型肝炎患儿血清蛋白质指纹图谱,以发现二者的差异蛋白质分子。方法收集60日龄前就诊、经手术证实为胆道闭锁和经过治疗、随访排除胆道闭锁的其他淤胆型婴儿肝炎患儿各21例的血清,采用弱阳离子交换蛋白质芯片（WCX2）为检测介质,经表面加强激光解离-飞行时间-质谱（SELDI-TOF-MS）测定,得到蛋白质指纹图谱,并运用BioMarker Wizard软件分析比较。结果在质荷比（m/z）1500～30000,检测出的25个蛋白峰中,强度变异系数15%;比较二者血清蛋白质指纹图谱,发现质荷比为8677的蛋白质峰在胆道闭锁患儿组下调（P〈0.05）。该蛋白质的减少可能在胆道闭锁的形成中起到一定作用,可能是胆道闭锁的蛋白质分子标志。根据发现的差异蛋白的分子质量及等电点,在Swiss蛋白质数据库中检索,发现与载脂蛋白AⅡ非常接近。结论SELDI-TOF-MS是一种快速、简便和高通量的分析方法,重复性好,与主要针对大分子质量的双相电泳研究在技术上形成互补,能直接筛选出胆道闭锁与淤胆型肝炎患儿的血清蛋白质指纹图谱中的差异蛋白质分子,可望用于胆道闭锁的早期鉴别诊断。
Objective To investigate the serum proteome fingerprint between biliary atresia and infant cholestatic hepatitis,and find serum proteomic difference between 2 diseases. Methods Serum samples were collected from 21 cases of biliary atresia and 21 cases of infant cholestatic hepatitis. All these children were within the first 60 days of life. Special serum protein or peptide fingerprint was determined by surface enhanced laser desorption/ionization - time of flight - mass spectrometry（ SELDI - TOF - MS） measurement after treating the sample onto weak cation exchange （WCX2） protein chip for each case. The obtained data were analyzed by BioMarker Wizard software. Results Within the range from 1 500 to 30 000 at the m/z value ,25 protein features were stably detected in the serum protein m/z pattern. One protein peak with the m/z value of 8 677 was statistically significant difference between 2 disease （P 〈 0.05 ）, it was downregulated in the group of biliary atresia. Decrease of this protein probably related to the pathogenesis of biliary atresia, and this low milecular weight protein maybe proteomic biomarker of this condition. We retrieved the differential protein in Swiss proteomie database according to its molecular weight and isoelectric point, found the differential protein was similar to APO AⅡ. Conclusions SELDI - TOF - MS ProteinChip technology is a quick, easy, convenient, and high output analyzing method that it is capable of selecting several relatively specific, potential biomarkers of disease. The abihty of SELDI - TOF - MS to resolve low molecular weight proteins suggest that the method may provide a mean of deciphering the biomarker - rich low milecular weight region of the human proteome. SELDI - TOF - MS is complementary to that of two - dimensional polyacrylamide gel electrophoresis （2 - D - PAGE）. Statistical difference exists in serum proteins between biliary atresia and infant cholestatic hepatitis. This study provide basis of unraveling the pathogenesis and finding potential biomarker for early diagnosis of biliary atresia.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2008年第7期501-503,共3页
Journal of Applied Clinical Pediatrics
基金
国家自然科学基金项目资助(30772271)
关键词
胆道闭锁
表面加强激光解离-飞行时间-质谱
蛋白质阵列分析
biliary atresia
surface enhanced laser desorption/ionization - time of flight - mass spectrometry
protein array analysis
