CAJ | 학술논문

目的：评估膝关节骨性关节炎(knee osteoarthritis，KOA)患者关节滑液双向凝胶电泳几种预处理方案的优劣。方法收集2012年1月-2012年3月在解放军总医院行关节镜诊治KOA患者关节滑液样本8例，混合均匀并等分为36份。9份不进行任何处理，9份用透明质酸酶处理，9份采用亲和层析法除高丰度蛋白，9份用透明质酸酶联合亲和层析法进行处理，对36份滑液进行双向凝胶电泳，比较未处理组与处理组以及处理组之间电泳图谱的差异并确定最佳预处理方案，采用最佳方案对12例不同严重程度KOA患者关节滑液进行预处理，分析寻找差异蛋白点并用质谱定性。结果采用透明质酸酶联合亲和层析法对样本进行预处理后获得的电泳图谱质量最佳。用此法进行预处理的12例不同严重程度KOA患者滑液，鉴定出了载脂蛋白、纤维蛋白原D片段等与关节疾病相关的蛋白因子。结论透明质酸酶联合亲和层析预处理方案能获得高质量的电泳图谱，可作为蛋白质组学研究中优先选取的样本预处理方案。
목적：평고슬관절골성관절염(knee osteoarthritis，KOA)환자관절활액쌍향응효전영궤충예처리방안적우렬。방법수집2012년1월-2012년3월재해방군총의원행관절경진치KOA환자관절활액양본8례，혼합균균병등분위36빈。9빈불진행임하처리，9빈용투명질산매처리，9빈채용친화층석법제고봉도단백，9빈용투명질산매연합친화층석법진행처리，대36빈활액진행쌍향응효전영，비교미처리조여처리조이급처리조지간전영도보적차이병학정최가예처리방안，채용최가방안대12례불동엄중정도KOA환자관절활액진행예처리，분석심조차이단백점병용질보정성。결과채용투명질산매연합친화층석법대양본진행예처리후획득적전영도보질량최가。용차법진행예처리적12례불동엄중정도KOA환자활액，감정출료재지단백、섬유단백원D편단등여관절질병상관적단백인자。결론투명질산매연합친화층석예처리방안능획득고질량적전영도보，가작위단백질조학연구중우선선취적양본예처리방안。
Objective To assess the advantages and disadvantages of several synovial fluid preparation methods for two-dimensional gel electrophoresis in patients with knee osteoarthritis (KOA). Methods Eight synovial fluid samples from KOA patients admitted to our hospital from January 2012 to March 2012 for arthroscopic diagnosis and treatment were mixed up and divided into 36 equal shares. Of which, 9 were not pretreated, 9 were treated with hyaluronidase, 9 with affinity chromatography to deplete high-abundance proteins, and 9 with combined hyaluronidase and affinity chromatography. The 36 equal sample shares were treated by two-dimensional gel electrophoresis. Different gel electrophoresis images of the treated and untreated samples were compared to determine the optimal pretreatment plan which was used in pretreatment of synovial fluid from 12 KOA patients. Differentially expressed proteins were detected and identified by mass spectrometry. Results The samples showed the best gel electrophoresis images, and the joint disease-related protein factors apolipoprotein and fibrinogen fragment D were identified in the synovial fluid from 12 KOA patients after the samples were treated with combined hyaluronidase and affinity chromatography. Conclusion Samples pretreated with combined hyaluronidase and affinity chromatography display high-quality gel electrophoresis images, and can thus be used as a first choice of pretreatment in proteomic study.