盆腔器官脱垂患者阴道组织比较蛋白质组学研究

目的:建立盆腔器官脱垂(pelvic organ prolapse,POP)患者及非脱垂对照者阴道组织蛋白质双向凝胶电泳图谱,并结合质谱技术分析其差异表达蛋白质,寻找POP疾病相关蛋白,以进一步阐明POP的发病机制。方法:收集绝经后年龄相匹配的POP组和对照组尿道周围阴道壁组织标本各10份,提取组织总蛋白,进行双向凝胶电泳(two-dimensional gel electro-phoresis,2-DE)得到凝胶图谱,采用Image Master 5.0 2D图象分析软件进行匹配和差异分析,识别两组间差异表达蛋白。将部分差异蛋白点胶内原位酶解后进行MALDI-TOF-MS分析,获取肽质量指纹图谱并进行生物信息学检索鉴定蛋白质。结果:得到了分辨率高,重复性好的POP组和对照组阴道壁组织蛋白双向凝胶电泳图谱,POP组的平均蛋白质点数为(871±96)个,正常对照组为(856±89)个。通过比较分析,差异表达蛋白质点数为27个。选取其中8个具有明显差异的蛋白点进行质谱鉴定,初步鉴定出5种蛋白质,其中2个蛋白质在POP组表达上调,3个蛋白质表达下调。结论:两者间存在一些差异表达的蛋白质,为阐明POP发病机制打下了坚实的基础。

Investigation on comparative proteomics of vaginal tissues of patients with pelvic organ prolapse

Objective:To establish the two-dimensional gel electrophoresis(2-DE) patterns of vaginal tissue protein from women with pelvic organ prolapse(POP) and women without POP,analyze the differentially expressed proteins in vaginal tissues by mass-spectrometric technique and find out the POP-related proteins to expound the pathogenesis of POP.Methods:The samples of periurethral vaginal wall tissues of postmenopausal,age-matched POP group and control group were collected,total tissue proteins were extracted to conducted two-dimensional gel electrophoresis,then gel electrophoresis patterns were obtained,and matched-analysis and differential-analysis were used by Image Master 5.0 2D image analysis software to find out the differentially expressed protein spots between the two groups.Some of the differentially expressed protein spots were in-gel digested and identified by MALDI-TOF-MS to obtain peptide mass fingerprint(PMF).Proteins were identified by PMF and database searching.Results:The two-dimensional gel electrophoresis patterns of vaginal wall tissues from POP group and control group with high resolution and good repeatability were obtained.The average protein spots of POP group and control group were(871±96) and(856±89),respectively.There were 27 differential protein spots between the two groups,eight of which were performed for mass-spectrometry and 5 kinds of proteins were obtained.2 kinds of proteins were upregulated and 3 kinds of proteins were downregulated in POP group.Conclusion:The differentially expressed proteins can be observed between POP group and control group,which can be used to elucidate the pathogenesis of POP for further study.