Biological Function Alternations With Stretch In Vaginal Wall Fibroblasts From Patients With Pelvic Floor Dysfunction

Objective:1 To find out the expression of typeⅠcollagen(colⅠ) , typeⅢcollagen(colⅢ),transforming growth factor beta1(TGF-β1)and Matrix metalloproteinase -1(MMP-1) in vaginal wall fibroblasts obtained from patients with pelvic floor dysfunction(PFD) under stretch.thus to explore the possible mechanism of PFD.2 Changes of the extracellular matrix to explore the pathogenesis of PFD.3 Explore pelvic floor dysfunction disease pathogenesis from the level of cell mechanics.Method:1 Study object: Twelve patients with PFD who underwent pelvic reconstructi- ve surgery in the second hospital of Hebei Medical University from October 2009 to June 2010 were recruited in the study.All the patients were examined and their prolapse were scored according to the International Continence Society Pelvic Organ Prolapse Quantitation (POP-Q) Classifi- cation, the patients were found in at least one part of POP-Q classification of gradeⅢ-Ⅳ(patients with uterine and vaginal anterior prolapse were three, patients with uterine and vaginal posterior prolapse were five ,patients with vaginal anterior and posterior prolapse were seven.), all patients with stress urinary incontinence were three, prolapse without stress incontinence were nine .The average age is 58.3±9.6.,The average parity is2.9±1.2. All the patients had been excluded connective tissue diseases such as respiratory system,cardiovascular system and the cutis diseases. Specimens were taken by the patient's consent, and signed informed consent.2 The specimens were taken and cultured in vitro, cells were identified as the vaginal wall fibroblasts,then sent to the Biomechanics Laboratory. the vaginal wall fibroblasts were cultured in monolayer on flexible substrates and subjectd to stretch 12% with sinusoidal waveforms at 0.3HZ by using FX-4000TM unit , The vaginal wall fibroblasts After stretch in 4 hours,15 hours ,24 hours as experimental group,with vaginal wall fibroblasts in static conditions as a control.3 Total RNA was extracted from fibroblasts and gene expression of colⅠ, colⅢ,TGF-beta1 and MMP-1 were measured before stretch applied and after4 hours,15 hours and 24 hours with stretch.the data were analysised with SPSS 13.0.Two-tailed test with P﹤0.05 were considered significant. Result:1 The cultured cells were freed from the tissue edge in 9-10 days after culture inoculation, cells were spindle-shaped, clear boundary, About 30 days Fibroblasts fusion after eliminate tissue, the fibroblasts were cord-like and whirlpool. Then immunocytochemical staining against vimentin positive expression rate is 91.6%, with drawn parts, confirmed cells were the vaginal wall fibroblasts.2 Primary cultured vaginal wall fibroblasts were cultured in monolayer on flexible substrates and subjectd to stretch 12% with sinusoidal waveforms at 0.3HZ by using FX-4000TM unit,with the time increasing,fibroblasts morphology and arrangement were changed,their arrangement becoming orderly manner.3 The expression of colⅠ,colⅢ,TGF-beta1 and MMP-1 were 1.178±0.071,0.673±0.029,0.852±0.039 and 1.167±0.036 in control, The expression of colⅠwere 1.092±0.038,1.198±0.031 and 1.120±0.098 after 4 hours,15 hours and 24 hours with stretch. The expression of colⅢwere 0.313±0.019, 0.504±0.033 and 0.853±0.210 after 4 hours,15 hours and 24 hours with stretch. The expression of TGF-beta1 were 0.673±0.052,0.897±0.051 and 0.895±0.015 after 4 hours,15 hours and 24 hours with stretch. The expression of MMP-1 were 0.669±0.109,1.085±0.176 and 1.325±0.048 after 4 hours,15 hours and 24 hours with stretch. Significant difference was observed between them.Conclution:1 the vaginal wall fibroblasts with PFD was successed created in vitro.2 the vaginal wall fibroblasts with PFD after mechanical stretch, the expression of colⅠ,colⅢ,TGF-beta1 and MMP-1were changing.which may lead to the extracellular matrix remodeling in pelvic connective tissue and play an important role in the occurrence of PFD.