中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2010年
45期
3193-3197
,共5页
肖军%史占军%吴志宏%梁锦前%赵赞栋%周亚鹏%兰天%邱贵兴
肖軍%史佔軍%吳誌宏%樑錦前%趙讚棟%週亞鵬%蘭天%邱貴興
초군%사점군%오지굉%량금전%조찬동%주아붕%란천%구귀흥
骨关节炎%软骨细胞%免疫电泳,双向%差异表达蛋白
骨關節炎%軟骨細胞%免疫電泳,雙嚮%差異錶達蛋白
골관절염%연골세포%면역전영,쌍향%차이표체단백
Osteoarthritis%Chondrocytes%Immunoelectrophoresis,two-dimensional%Differential expressed proteins
目的 探讨骨关节炎(OA)差异表达蛋白的病理作用.方法 分别从正常膝关节提取正常软骨(NAC,n=22)和从接受膝关节表面置换的OA患者提取OA软骨(OAC,n=17).分离软骨细胞,提取总蛋白直接进行双向电泳分析.胶图导入Imagemaster 2-D platinum3.0软件筛选差异表达蛋白点,切胶消化并提交质谱分析.质谱生成的肽段指纹序列输入到MASCOT,依托NCBI蛋白数据库确立差异表达蛋白的身份.结果 NAC和OAC组双向电泳胶图分别显示约1000个蛋白点,其中35个NAC蛋白点和31个OAC蛋白点被确立为差异表达蛋白.通过质谱分析识别19个差异表达蛋白,包括6型胶原、与软骨细胞胶原代谢相关的酶、应激相关蛋白,以及功能未知蛋白.结论 差异表达蛋白的分离和识别提示了多方面的OA病理信息,为深入认识OA的分子病因学拓宽了视野.
目的 探討骨關節炎(OA)差異錶達蛋白的病理作用.方法 分彆從正常膝關節提取正常軟骨(NAC,n=22)和從接受膝關節錶麵置換的OA患者提取OA軟骨(OAC,n=17).分離軟骨細胞,提取總蛋白直接進行雙嚮電泳分析.膠圖導入Imagemaster 2-D platinum3.0軟件篩選差異錶達蛋白點,切膠消化併提交質譜分析.質譜生成的肽段指紋序列輸入到MASCOT,依託NCBI蛋白數據庫確立差異錶達蛋白的身份.結果 NAC和OAC組雙嚮電泳膠圖分彆顯示約1000箇蛋白點,其中35箇NAC蛋白點和31箇OAC蛋白點被確立為差異錶達蛋白.通過質譜分析識彆19箇差異錶達蛋白,包括6型膠原、與軟骨細胞膠原代謝相關的酶、應激相關蛋白,以及功能未知蛋白.結論 差異錶達蛋白的分離和識彆提示瞭多方麵的OA病理信息,為深入認識OA的分子病因學拓寬瞭視野.
목적 탐토골관절염(OA)차이표체단백적병리작용.방법 분별종정상슬관절제취정상연골(NAC,n=22)화종접수슬관절표면치환적OA환자제취OA연골(OAC,n=17).분리연골세포,제취총단백직접진행쌍향전영분석.효도도입Imagemaster 2-D platinum3.0연건사선차이표체단백점,절효소화병제교질보분석.질보생성적태단지문서렬수입도MASCOT,의탁NCBI단백수거고학립차이표체단백적신빈.결과 NAC화OAC조쌍향전영효도분별현시약1000개단백점,기중35개NAC단백점화31개OAC단백점피학립위차이표체단백.통과질보분석식별19개차이표체단백,포괄6형효원、여연골세포효원대사상관적매、응격상관단백,이급공능미지단백.결론 차이표체단백적분리화식별제시료다방면적OA병리신식,위심입인식OA적분자병인학탁관료시야.
Objective To investigate the molecular mechanism of osteoarthritis (OA) through the differential expressed proteins of advanced osteoarthritic chondrocytes.Methods Normal articular cartilage (NAC,n = 22) were obtained from knee cartilage of donors without knee diseases and osteoarthritic cartilage (OAC,n = 17) from OA patients undergoing knee arthroplasty.Total protein was extracted from chondrocytes and loaded directly for two-dimensional gel electrophoresis (2-DE).Gel images were analyzed with the software Imagemaster 2-D platinum3.0 to screen the differential expression protein spots.Target spots were excised and digested and the resulting peptides submitted for mass spectrometry analysis.Identities of differential expressed proteins were recognized through matching the peptide mass finger printings of each spot with NCBI protein database by MASCOT.Results About 1000 protein spots were presented in the 2-DE gels for each group.Thirty-five NAC spots and 31 OAC spots were confirmed to be differential expression protein spots.Among these spots,19 proteins were identified by MASCOT,including type Ⅵ collagen,enzymes involved in collagen synthesis,shock related proteins and some novel proteins with unknown functions.Conclusion The differential expressed proteins isolated and identified by the present study provide valuable information on various aspects of OA pathology.It may help us to gain new insights into the molecular mechanism of OA.
