CAJ | 학술논문

目的：建立一个良好的粪便蛋白质提取、分析和鉴定的方法，为进一步探索消化系统疾病无创性诊治靶标提供新思路和奠定良好研究体系。方法采用生理盐水、Tris-HCl 和尿素缓冲液三种粪便蛋白质提取体系，处理健康人群、慢性萎缩性胃炎、胃腺癌术前、胃腺癌术后的粪便标本，通过 SDS-PAGE 进行比较，获得较好的粪便蛋白质提取方法。再利用液相串联质谱（1D LC-MS／MS）对提取的蛋白质进行初步分析。结果生理盐水和 Tris-HCl 缓冲液提取粪便蛋白质的效率明显较尿素法高，损失少，是粪便蛋白质比较有效和合理的提取方法。不同人群来源的粪便中均含有丰富的蛋白质，正常人、胃腺癌手术前及术后鉴定到的蛋白质达一百多种，在组成和含量上个体差异明显。疾病与正常间、疾病治疗前后，粪便蛋白质指纹图谱变化大。胃腺癌术前粪便中含有大量的免疫球蛋白和角蛋白，同时也鉴定到了α1-抗胰蛋白酶，但在其术后和健康者粪便中未见到这一现象。结论正常与胃癌、胃癌手术前与术后粪便蛋白质不仅有蛋白质种类的差异，而且相同蛋白质量的差异也相当明显。因此，利用蛋白质组学技术筛选人粪便中胃癌相关的蛋白质标志物有较强的可行性，但在未来的研究中需要差异分析结合定量质谱的方法进行。
목적：건립일개량호적분편단백질제취、분석화감정적방법，위진일보탐색소화계통질병무창성진치파표제공신사로화전정량호연구체계。방법채용생리염수、Tris-HCl 화뇨소완충액삼충분편단백질제취체계，처리건강인군、만성위축성위염、위선암술전、위선암술후적분편표본，통과 SDS-PAGE 진행비교，획득교호적분편단백질제취방법。재이용액상천련질보（1D LC-MS／MS）대제취적단백질진행초보분석。결과생리염수화 Tris-HCl 완충액제취분편단백질적효솔명현교뇨소법고，손실소，시분편단백질비교유효화합리적제취방법。불동인군래원적분편중균함유봉부적단백질，정상인、위선암수술전급술후감정도적단백질체일백다충，재조성화함량상개체차이명현。질병여정상간、질병치료전후，분편단백질지문도보변화대。위선암술전분편중함유대량적면역구단백화각단백，동시야감정도료α1-항이단백매，단재기술후화건강자분편중미견도저일현상。결론정상여위암、위암수술전여술후분편단백질불부유단백질충류적차이，이차상동단백질량적차이야상당명현。인차，이용단백질조학기술사선인분편중위암상관적단백질표지물유교강적가행성，단재미래적연구중수요차이분석결합정량질보적방법진행。
Objective To set up a method of stools protein extraction,analysis and identification in order to get the new nonin-vasive indicators of human digestive diseases.Methods The stools proteins,collected from healthy persons,the patients with atrophic gastritis,those who suffed from gastric carcinoma and postoperative patients with gastric carcinoma respectively, were extracted in three different ways including saline,Tris-HCl buffer and Urea buffer,the best way was selected by using SDS-PAGE,then a preliminary analysis of stools proteins was performed by 1D LC-MS/MS.Results The methods of saline and Tris-HCl buffer could get more stools proteins than the method of urea.The proteins in stools from the healthy persons, the patients with atrophic gastritis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma were all abundant and more than one hundred.There was a significant difference in stools protein maps among the various populations.Alpha1-antitrypsin,a number of immunoglobulin and keratin were identified in the stools from patients with gastric carcinoma but not postoperative patients with gastric carcinoma and the healthy persons.Conclusions In this re-search,there was a significant difference in stools protein maps among the healthy persons,the patients with atrophic gastri-tis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma,not only the composition of stools proteins,but also the abundance of same proteins.Therefore,using proteomics technologies to screening of the noninvasive indicators in human stools is viable.The study recommended that the noninvasive indicators in human stools should be iden-tified with quantitative differences analysis combination of quality of mass spectrometer method in the future research.