摘要目的 应用基因表达谱芯片检测绝经后盆腔器官脱垂(POP)患者与绝经后非POP患者子宫主韧带组织中差异表达的基因,探讨POP发生的分子机制.方法 选择2007年1-5月在北京协和医院妇产科手术的绝经后POP患者3例为POP组,同期因妇科良性疾病行子宫全切除术的绝经后非POP患者3例为对照组.获取两组患者的子宫主韧带组织,进行组织来源确定及形态学观察;应用基因表达谱芯片筛选两组子宫主韧带组织中的差异表达基因,并进行功能分析;采用实时荧光定量PCR技术验证所筛选的差异表达基因的表达水平.结果 POP组患者子宫主韧带组织中胶原纤维与平滑肌束失去正常的排列,形态紊乱,可见胶原纤维断裂,平滑肌束细碎.采用基因表达谱芯片共筛选出179个差异表达的基因,其中包括加个功能未知的基因.107个基因在POP组中表达上调,72个基因在POP组中表达下调.差异基因涉及多种功能蛋白和代谢通路,其中Wnt信号传导通路的变化最显著(P=0.000).实时荧光定量PCR技术检测证实,COL1A1、DKK1、SFRP1、FZD5、WNT16b基因在POP组中的表达水平分别为2.98±1.40、3.03±0.48、8.13±4.42、5.19.4±3.50和12.40±3.88,均明显高于对照组(分别为1.09±0.08、1.20±0.18、0.41±0.51、0.87±0.24和1.40±0.47,P<0.05),与基因表达谱芯片的检测结果一致.结论 POP的病理牛理过程复杂,涉及多种功能蛋白和代谢通路,其中Wnt信号传导通路拮抗剂DKK1、SFRP1介导的神经退行性病变可能与POP的发病密切相关.

abstractsObjective To identify the differentially expressed genes in cardinal ligament between patients with pelvic organ prolapse ( POP) and postmenopausal women without POP by Human Genome Expression Chip and explore the potential molecular mechanism involved in POP.Methods From January to May,2007,cardinal ligament samples were obtained from 3 postmenopausal patients with POP-Q stage Ⅲ and 3 postmenopausal patients underwent hysterectomy due to other benign gynecologic diseases without POP in Peking Union Medical College Hospital.HE and Masson's trichrome staining was used to verify tissue origin and inspect histological changes.Those differentially expressed genes in cardinal ligaments were identified by Human Genome Chip and further interrogated with Gene Ontology (GO) and Pathway Analysis.Those remarkable expressed genes were confirmed by qRT-PCR.Results Alterations of ligament architecture in POP patients included disarrangement and collapse of smooth muscle bundles and collagen fibers.A total of 179 differentially expressed genes were screened between POP and non-POP cardinal ligament tissue,including 20 functional unknown genes.A total of 107 genes were upregulated in POP group,while 72 genes downregulated.Those differentially genes were revealed associated with multiple functional proteins and metabolic pathways by biological analysis.Among these,Wnt signaling pathway exhibited the most remarkable changes.Real-time quantitative PCR showed the genes of COL1Al,DKK1,SFRP1,FZD5,WNT16b in POP group (2.98 ±1.40,3.03 ±0.48,8.13 ±4.42,5.19 ±3.50,12.40± 3.88) were upregulated significantly compared with non-POP group (1.09 ±0.08,1/20 ±0.18,0.41 ± 0.51,0.87 ±0.24,1.40 ±0.47; P < 0.05 ).Conclusions The pathophysiology of POP is complex and associated with multiple functional proteins and metabolic pathways.Among these,the antagonist DKK1,SFRP1 in Wnt signaling pathway may contribute to a neurodegenerative role in POP development.