CAJ | 학술논문

目的在已有实验基础上进一步建立优化的用于人胰液差异蛋白质组研究的双向凝胶电泳(two-dimensional electrophoresis,2-DE)方法;探讨利用混合胰液进行胰腺疾病的胰液差异蛋白质组学的可行性.方法通过ERCP术中放置鼻胰管引流收集5例胰腺癌(其中2例手术病理分别为胰腺中分化导管腺癌及导管内乳头状瘤癌变,另外3例经影像学检查结合临床诊断)和6例慢性胰腺炎胰液标本,取其中一例慢性胰腺炎胰液标本,通过改进胰液标本的处理方法、泡胀液的组成和一向等电聚焦条件后进行双向凝胶电泳,并和先前实验条件下的胰液2-DE图谱进行比较分析;同时利用优化后的双向凝胶电泳方法比较单份胰液和同病种混合胰液标本的2-DE图谱.结果应用优化的2-DE方法,在胰液蛋白质加样量为200 μg时,可见凝胶上约有280个蛋白质点,有较好分辨率,较原条件下电泳图谱有较大改进.单份胰液和同病种混合胰液标本的2-DE图谱有较高程度的相似性(>75%).结论优化后的胰液2-DE方法切实可行,通过各不同胰腺疾病混合胰液的2-DE图谱差异比较可为胰腺疾病的胰液差异蛋白质组学研究建立良好的基础.
목적 재이유실험기출상진일보건립우화적용우인이액차이단백질조연구적쌍향응효전영(two-dimensional electrophoresis,2-DE)방법;탐토이용혼합이액진행이선질병적이액차이단백질조학적가행성.방법 통과ERCP술중방치비이관인류수집5례이선암(기중2례수술병리분별위이선중분화도관선암급도관내유두상류암변,령외3례경영상학검사결합림상진단)화6례만성이선염이액표본,취기중일례만성이선염이액표본,통과개진이액표본적처리방법、포창액적조성화일향등전취초조건후진행쌍향응효전영,병화선전실험조건하적이액2-DE도보진행비교분석;동시이용우화후적쌍향응효전영방법비교단빈이액화동병충혼합이액표본적2-DE도보.결과 응용우화적2-DE방법,재이액단백질가양량위200 μg시,가견응효상약유280개단백질점,유교호분변솔,교원조건하전영도보유교대개진.단빈이액화동병충혼합이액표본적2-DE도보유교고정도적상사성(>75%).결론 우화후적이액2-DE방법절실가행,통과각불동이선질병혼합이액적2-DE도보차이비교가위이선질병적이액차이단백질조학연구건립량호적기출.
Objective To establish an optimal method of two-dimensional gel electrophoresis (2-DE) for proteomic analysis of human pancreatic juice (pj) on the basis of former work and explore the feasibility of differential proteomic analysis of human pancreatic juice by the use of mixed pancreatic juice samples. Methods Pj samples from 5 pancreatic cancer patients (two cases were pathologically diagnosed moderately differentiated adenocarcinoma and intraduetal papilloma canceration respectively,three cases were diagnosed by inageology and clinical manifestations) and 6 chronic pancreatitis pa-tients were included in this study. Pj samples were collected by naso-pancreatic drainage during endo-scopic retrograde cholangiopancreatography. 2-DE was conducted on one pj sample of chronic panerea-titis by improving sample preparation techniques. Compositions of rehydration solutions and methods of isoelectric focusing and 2-DE maps were compared with those got from former 2-DE method. 2-DE method and 2-DE maps of single pj sample and mixed pj samples of the same pancreatic disease were also compared by the optimized method of 2-DE. Results With the improved 2-DE method and when 200 μg protein of pancreatic juice was loaded, about 280 protein spots could be successfully separated with better differentiation rate as compared with the former method. There was higher similarity (>75%)between the 2-DE maps of single pj sample and mixed pj samples of the same pancreatic disease. Conclusion Optimized method of 2-DE has reliable effect and the comparison of 2-DE maps of mixed pj samples of different pancreatic diseases would become the basis of differential proteomic analysis of human pancreatic juice.