Effect Of 17-β Estradiol In Cultured Vaginal Wall Fibroblasts From Womem With Pelvic Floor Dysfunction Before And After Stretch

Objective:To study the differences expression of CollagenⅠ, CollagenⅢ, lysyl oxidase like 1(LOXL1 , fibrinectin(FN)before and after stretch in the pelvic function dysfunction(PFD)patients vaginal wall fibroblasts cultured in vitro,and the effect in PFD pathogenesis.Methods :1 Study object: From October 2009 to September 2010,we collected vaginal wall organization of the patients who were suffering from pelvic function dysfunction disease in the second Hospital, Hebei Medical University. All patients scored to quantify according to International Continence Society published the pelvic organ prolapse quantitative examination(POP-Q) in 1996,and at least one organ on the stages were III or IV.The average ages were 58.3±9.6. All patients were not used hormone drugs before surgery 3 months,and were not merged connective tissue diseases. Informed consent was obtained from all recruited subjects.2 Fibroblast cell culture: Use of tissue and collagenase digestion method for primary culture in the vaginal wall fibroblasts,take 3 to 5 generations fibroblasts on experiment. Rates of cell proliferation of fibroblasts were examined by MTT assay.3 Fibroblast stretch: Application Flexcercell 4000 stretch systems to cell, after stretch,with different concentrations of 17-βestradiol cultured fibroblasts for 48 hours,without drug group as control group.4 Reverse transcription polymerase chain reaction (RT-PCR):extraction mRNA of the cells,RT-PCR,electrophoresis,photograph and observed the expression and record results in each group.Results:1 Situation of cell culture: After the success of primary culture conditions, passaged with 1:2 ratio of sub-bottle.Cells were identified by immuno- histochemisty,vimentin positive,keratin and actin negative were identified as fibroblasts.2 Situation of cell proliferation: Before stretch,the proliferation rates of vaginal wall fibroblasts from low to high concentrations of 17-βestradiol were0.23,0.38,0.38,0.54.After stretch were 0.06,0.07,0.10,0.17.Between the two groups was statically significant(P<0.05). The proliferation rates increased with the concentration increasing.3 The expression of each factor before and after stretch:①The expression of CollagenⅠwere 1.1872±0.0733,1.5035±0.0733,(P<0.05).②CollagenⅢwere 0.7229±0.0966, 0.2024±0.0465,(P<0.05).③LOXL-1 were 0.7724±0.1873,1.0855±0.0805,(P<0.05).④FN were 0.4290±0.1168, 0.4215±0.0830,(P>0.05).4 The expression of each factor in high drug concentrations of 17-βestradiol: CollagenⅠwere 3.0809±0.1862;CollagenⅢwere 1.3032±0.2994; LOXL-1 were 1.5863±0.3241; FN were 1.1418±1.0030.Compared with other groups was statistically significant(P<0.05).Conclusion:1 Stretch inhibited the proliferation of cultured PFD vaginal wall fibroblasts in vitro. 17-βestradiol promoted the proliferation,and cell proliferation rates increased with the concentration increasing. To a certain extent ,17-βestradiol could reversed the stretch on the inhibition of cell proliferation .2 The vaginal wall fibroblasts of the pelvic function dysfunction (PFD)patients in vitro,the expression of CollagenⅠand LOXL-1 increased, CollagenⅢdecreased ,and FN did not change. 17-βestradiol increased synthesis of extracellular matrix components from fibroblasts after stretch.3 Stretch and estrogen may play an important role in PFD pathogenesis.