阴道前壁成纤维细胞参与女性压力性尿失禁形成的microRNA-靶基因-蛋白质级联调控机制探讨

Occurrence of stress urinary incontinence due to the weakened urinary tract support, which results from the defect in supporting structures of anterior vaginal wall. Epidemiological studies report that 20% adult women and up to 50% of women over the age of 60 have symptoms of stress urinary incontinence (SUI), which is a major health and quality-of-life issue for women. Nerve injury-degeneration mechanism and abnormal extracellular matrix metabolism mechanism is concerned currently. And, in which, the genetic factor plays an important role. But the specific molecular pathogenesis is not yet clear. To date, there are no published researches on the relationship between microRNA and SUI. Our previous study established microRNA differential expression profile at histological level, and their involvement in neural damage mechanism has also been preliminary investigated. The supporting structures of urinary tract are extracellular matrix of anterior vaginal wall, which are synthesized by fibroblast mainly. Therefore, fibroblast will act as the research subject in this study. Firstly, primary cell culture of anterior vaginal wall fibroblast will be done. Candidate target genes and microRNAs are obtained through integrating bioinformatic database with results from Gene Chips. Further, functional study of microRNAs will be conducted through its overexpression and inhibition test. Finally, luciferase reporter gene assay system will be used to verify the target genes. We look forward to establish holonomic differential microRNA expression profile (including cellular and histological level), establish the microRNA -mRNA-protein cascade regulating mechanism, and discover the candidate target of genetic intervention, as well as provide a molecular theoretical basis for fibroblast as seed cells in tissue engineering therapy for SUI.

约近50%的老年妇女患病率使得压力性尿失禁（SUI）成为突出的医疗问题和备受关注的社会问题。SUI发病机制尚不明了，神经变性与细胞外基质代谢异常学说备受关注，遗传因素在其中发挥着重要作用。迄今，尚无microRNA（miRNA）与SUI的相关研究报道。我们的前期研究建立了SUI组织水平的miRNA差异表达谱，并对神经损伤机制做了初步探讨。本项目拟以原代培养的阴道前壁成纤维细胞为研究对象，结合前期研究，采用“miRNA芯片-mRNA芯片”数据整合、构建过表达载体及抑制性反义核酸分子转染细胞进行miRNA功能研究、荧光素酶双报告基因检测系统验证靶基因的路线，从整体上（包括组织水平与细胞水平）建立miRNA差异表达谱，从细胞外基质代谢角度探寻SUI病理过程中miRNA-靶基因-蛋白质级联调控机制，为SUI基因干预提供候选靶点，为成纤维细胞作为SUI组织工程治疗的种子细胞提供分子水平上的理论依据。