盆腔器官脱垂发病机制的比较蛋白组学研究  

Proteomic analysis of cardinal-sacral ligament in women with or without pelvic organ prolapse

作　　者：王静怡[1] 孙智晶[1] 朱兰[1] 郎景和[1] 梁硕[1] 

机构地区：[1]中国医学科学院北京协和医学院北京协和医院妇产科,北京100730

出　　处：《生殖医学杂志》2015年第6期481-486,共6页Journal of Reproductive Medicine

基　　金：国家青年自然科学基金:盆腔器官脱垂的比较蛋白组学研究(3090157)

摘　　要：目的应用双向荧光差异凝胶电泳和质谱技术,筛选并鉴定盆腔器官脱垂(POP)患者与正常对照之间的差异表达蛋白,探讨POP发生的分子机制。方法在绝经状态、年龄、体重指数、产次等因素匹配的前提下,选取5例行子宫全切和盆底重建的POP患者与5例无POP行子宫全切患者的主骶韧带复合体标本。提取组织总蛋白,普通双向电泳分离,银染显色鉴定蛋白质抽提质量和在胶图上的分布情况。荧光染料分别标记POP组、对照组以及内标,进行双向电泳得到双向荧光差异凝胶图谱。对胶图进行匹配和差异分析,筛选出两组之间丰度差异≥1.5,P<0.05的差异表达蛋白质点。将差异蛋白质点胶内酶解后进行质谱分析和数据库搜索,鉴定差异表达的蛋白质点。结果提取的组织蛋白经定量和普通双向电泳检测,平均可获得(1 224±22)个蛋白质点。获得5张双向荧光差异凝胶电泳胶图,每张胶图上共约(1 111±54)个蛋白质点,经过比较分析筛选出33个差异表达蛋白质点,均在POP组下调,最大下调比例为8.5倍。质谱成功鉴定出半凝乳素-1、亲环素A、丝切蛋白等蛋白,均在POP组下调。结论在POP患者中半凝乳素-1和亲环素A发生降调节提示,POP的发生可能与神经损伤相关。Objective：To identify the differentially expressed proteins in cardinal-sacral ligament between women with or without pelvic organ prolapse（POP）by 2-D fluorescence difference gel electrophoresis（2-D DIGE）and mass spectrometry for further exploring the molecular mechanism associated with POP.Methods：The cardinal-sacral ligament samples were obtained from five postmenopausal patients with POP-Q stage Ⅲ and five postmenopausal patients without POP,who were matched in age/BMI/parity.Total proteins were extracted and quantified,and then separated by two-dimensional gel and stained with silver.The samples were labeled with Cydye DIGE fluor and then separated by 2D-DIGE.The gels were scanned and analyzed.The spots with average differential rate（｜AV.Ratio｜）≥1.5and P〈0.05 were identified as differentially expressed proteins.The spots were picked and digested,and the peptides were identified by MALDI-TOF-MS.The peptide mass fingerprint（PMF）was searched in the database.Results：The high-quality proteins were extracted from the samples and well separated by 2Dgel.A total of 33 differentially expressed proteins were detected,all of which were down-regulated in patients with POP,and the biggest down regulation ratio is 8.5times.Galectin-1,cyclophilin A and cofilin-1were identified by MALDI-TOF-MS.They were down-regulated in POP group.Conclusions：Down-regulation of galectin-1and cyclophilin in the patients with POP suggests that development of POP is associated with neural injury.

关 键 词：盆腔器官脱垂 主骶韧带复合体 双向荧光差异凝胶电泳 神经变性 

分 类 号：R[医药卫生]