Research On Expression Of TGF-β1, Smad3and CTGF In The Incidence Of Women With Stress Urinary Incontinence

Stress Urinary Incontinence (SUI) is known as dysfunctional disease caused bythe dysfunction of urethra to control micturition. The dysfunction is mostly inducedby female pelvic floor muscle damage, flabby fascia, and anatomical urethra positionchange, due to childbirth, trauma and other factors. SUI is a complicated diseasetriggered by multiple factors, and is often accompanied by female pelvic organprolapse (POP). Its characteristic is that people with the disease have no leakage ofurine in normal times. However, when coughing, laughing, or taking heavy manuallabor, they have higher abdominal pressure, and the urine leakage arisesuncontrolledly. The severity of symptom increases as the age increases. Therefore,The International Continence Society defined SUI as: involuntary urine outflow withsudden increase of abdominal pressure, which is not caused by the tension of thedetrusor, but by the anatomical urethra abnormity of pelvic floor and imbalance ofpressure conduction. Someone refer the disease as "social cancer", that is becausealthough stress urinary incontinence does not play as cardiovascular diseases, tumorand so on, which seriously endanger people’s life and health, SUI brings in a series ofsocial, economic, and health problems, etc. With the advent of era of aging, the onsetof stress urinary incontinence morbidity rates increase significantly, so the influencehas been taken for granted to get more and more attentions. As "a hammockhypothesis" and "pelvic floor integral theory" put forward, people gradually realized that only by testing urethral pressure, urethral length and angle of bladder urethraanatomical changes is insufficient for explaining the pathogenesis of SUI completelyand reasonably. This forces the researchers to study the pelvic anatomical structureand function in the level of molecular biology. Numerous studies have shown thatwomen with SUI pelvic floor support structure of collagen and elastic protein contentis significantly reduced, resulting in less resilient support tissue and the presence ofpelvic floor relaxation. And transforming growth factor beta1(TGF-β1), as the mostimportant cytokines, is directly related to the synthesis and degradation of collagenand elastin, and it can also change the quantity and activity of inhibiting factor ofmatrix-degrading enzymes. This play a key role in the process repairmen for damagecaused in childbirth or surgery to pelvic support tissue. Connective tissue growthfacto（rCTGF）can stimulate fibroblast proliferation, secretion of collagen, especiallythe fibrosis after sexual disease, trauma, scarring. However, seldom studies have beenreported concerning pelvic floor disorder. TGF-β is the major growth factorimplicated in vascular matrix accumulation via the SMAD signal pathway; The Smad3protein, owning two-way regulation of cell function, involve in controlling thehealing of the wound tissue.To study the pathogenesis of female SUI, we use immunehistochemical andreverse transcription and polymerase method to detect TGF-β1, CTGF and Smad3expression in women vaginal wall tissue. By analyzing the correlation of the threefactors, we may demonstrate their roles in SUI mechanism, and provide theoreticalreferences for prevention and treatment of the disease.Materials and methods1. MaterialsWe randomly selected60cases of severe patients with stress urinaryincontinence from June2012to April2013in our hospital obstetrics and gynecologyhospital as the experimental group, including25cases of simple SUI,35cases of SUImerger with POP, with methods of operation method of transvaginal no middletension urethra suspension (trans-obturator vaginal tape, TVT-O), vaginal wall before and after repair, vaginal wall before the suspension. Meanwhile, in the sameperiod of20patients selected with cervical intraepithelial neoplasia is not with SUI ascontrol group, operation method for abdominal or vaginal hysterectomy. Three groupsin terms of age, production time, and body mass index (bmi) there was no statisticallysignificant difference (P>0.05), and have good comparability. All cases withoutrheumatoid arthritis, hyperparathyroidism, influence of collagen metabolismdisorders such as preoperative never use a sex hormone drugs within three months,who did not have functional ovarian tumors, no urinary tract infection, there is nohistory of previous pelvic surgery. SUI is diagnosed by the evaluation of the patient’smedical history, examination of department of gynaecology, pressure test, pressuretesting, the urine pad experiment and dynamic test results.2. MethodsBy immune histochemical method we detect the expression level of vaginal walltissue TGF-beta1, the expression of CTGF and Smad3protein in SUI group, SUI+POP group and the control group patients. Meanwhile, the technology of ReverseTranscription-Polymerase Chain Reaction (RT-PCR) is used to detect three groupsof TGF-beta1, CTGF and Smad3mRNA level.3. Statistical analysisUsing SPSS17.0statistical software for data processing. Rate and constituentratio are comparied by chi-square test, any two groups are comparied by using thechi-square segmentation method, Alpha=α/3=0.0167; Multiple sets of quantitativedata comparison are assessed by using single factor analysis of variance. Correlationanalysis is used to analyzed contingency related classification data. The inspectionlevel. Alpha=0.05. Results1. The immunohistochemical staining resultsThree groups of patients with vaginal wall in the organization are visible TGF-beta1protein, the expression of CTGF protein and Smad3protein, a pale yellow tobrownish yellow brown.TGF-beta1protein and CTGF protein is mainly expressedin the cell cytoplasm, a pale yellow to brownish yellow brown diffusedistribution;Smad3proteins expression mainly in the nucleus, and present a yellowishto brownish yellow brown granules of cells is positive.1) TGF-beta1protein expression within the three groups28%、17.14%、95%. Thedifference was statistically significant (chi-square=34.327, P <0.01), while the TGF-beta1protein expression in the SUI group and SUI+POP group has no statisticallysignificant difference (P>0.05).2) CTGF protein positive expression within the three groups16%、20%、95%. Thedifference was statistically significant (chi-square=37.717, P <0.01), while CTGFprotein in SUI group and SUI+POP group, the expression differences have nostatistical significance (P>0.05).3) Smad3protein positive expression within the three groups24%、14.29%、90%.The difference was statistically significant (chi-square=33.933, P <0.01), whileCTGF protein in SUI group and SUI+POP group, the expression differences have nostatistical significance (P>0.05).2. RT-PCR resultsSemi-quantitative determination results showed that TGF-beta1mRNA,CTGFmRNA and Smad3mRNA expression are all positive in the three groups.1) TGF-beta1mRNA expression in the SUI group,SUI+POP group and thecontrol group (0.294±0.013),(0.293±0.019),(0.610±0.052), the former expressiondecreased obviously, the difference was statistically significant (P <0.01), while SUIgroup compared with SUI+POP group, there was no statistical significance (P>0.05) difference expression.2) CTGFmRNA expression in the SUI group,SUI+POP group and the control group (0.085±0.012),(0.092±0.011),(0.272±0.029). The former expression decreasedobviously, the difference was statistically significant (P <0.01), while SUI groupcompared with SUI+POP group, there was no statistical significance (P>0.05)difference expression.3) Smad3mRNA expression in the SUI group and SUI+POP group and the controlgroup(0.125±0.014),(0.132±0.014),(0.327±0.047), the former expression decreasedobviously, the difference was statistically significant (P <0.01), while SUI groupcompared with SUI+POP group, there was no statistical significance (P>0.05)difference expression.3. The correlation analysisAccording to the contingency correlation analysis showed that SUI group ofvaginal wall tissue positive correlation between CTGF protein and TGF-beta1(r=0.959, P <0.05); SUI group of vaginal wall tissue TGF-positive correlation betweenCTGF expression and beta1(r=0.944, P <0.05); SUI group of vaginal wall tissueCTGF positive correlation between the expression of Smad3and (r=0.965, P <0.05).ConclusionsThe low expression of transforming growth factor beta1, connective tissuegrowth factor and Smad3may participate in the occurrence of SUI.