| Urine incontinence is a common symptom of female. There are many kinds of urine incontinence,such as true urine incontinence,functional urine incontinence, stress urine incontinence,urgent urine incontinence.Among the total,stress urine incontinence,which take an account of 50-70%,is the most common one.Stress urinary incontinence(SUI) is defined as the phenomenon when abdominal pressure increasing even when resting,bladder neck and urethral cann't maintain certain pressure and urine overflow. It seriously affect women's quality of life,with the aging population increase,SUI is more and more received the general scholars' attention.Scholars have taken an extensive research fundamentally on its mechanisms to prevent SUI occurs,but haven't obtained unanimous conclusion currently.SUI occurred closely related to the weakness of the pelvic support organization. Elastic protein(Elsatin), collagen, layer adhesion protein is main ingredient of extracellular matrix (ECM)'s. A study found that collagen content, structure, function, proportional anomalies ofⅠ/Ⅲcollagen type are closely related to SUI. Other studies have found that active fascia metabolic of pelvic support organizations and the increased decomposition of collagen might be involved in SUI formation. Elastin is also a main ingredient of pelvic support groups, it can make the organization has loft and reversible deformation ability. And it also has significant meaning to maintain strengthen the function of pelvic organizations. At present the Elastin has been reaserched in POP cause aspect somewhat, but in the SUI aspect has not been involved in at home and abroad.Matrix metalloproteinases(MMPs) is a group of important enzymes to adjust the metabolism of ECM and it plays a direct or indirect role in decomposed and anabolic aspects of pelvic support groups. MMPs is a group of protease family which contin Zn2+. It can degrade the extracellular matrix (ECM) and the basement membrane composition,its family includes the following five kinds of protease:(1) collagenase (MMP-1 and MMP-8), (2) membrane type protease (MT1-MMP, MT2-MMP etc.), (3) gelatinases (MMP-2 and MMP-9), (4) matrix dissolve hormone (MMP-3, MMP-7 and MMP-10), (5) elastase (MMP-12). Among them, the main substrate of MMP-12 is elastic fiber, layer adhesion protein and fibronectin etc. Now, a large studies had indicated that it is tending to high expression in patients with disease such as esophageal, epithelial oarian, lung and colon cancer. Because of vaginal wall tissue contains elastic fiber, presumably MMP-12 and female SUI pathogenesis also are linked.Matrix metalloproteinases inhibitors (inhibitors of tissue metalloproteinase, TIMPs) is a multigene family of coding protein.It have negative adjustment effect to MMPs. TIMP-1 is specific inhibitor to MMP-12.In physiological condition, they are both in a certain extent to keep balance which can make destruction and rebuilding of body elastin maintain normal state. When this balance is broken, a series of body pathological conditions can be caused. Therefore, imbalance between TIMPs and MMPs is also one of the reasons for the stress urine incontinence.ObjectiveThis topic take vaginal front wall organization of female as experimental subjects, using immunohistochemistry detecting content of MMP-12, TIMP-1, Elastin in each vaginal wall tissue, and analy the content of the correlation between the three indexes,And RT-PCR method to detect Elastin mRNA in each group.Understand relationship between MMP-12,TIMP-1, Elastin expressions and female stress urinary incontinence, in order to provides the basis to disease prevention and treatment in aspect of pathogeny in the future.Materials and Methods1 MaterialsAll patients are from The Third Affiliated Hospital of Zhengzhou University gynecological from 2009 August to 2010 February. With SUI patients 30 cases, Age between 37-60. POP patients 30 cases, Age between 40-59,and choose 30 patinents with cervical intraepithelial neoplasia or non-functional ovarian tumor as control group Age between 38-65.Selection standards:SUI groups:with SUI symptoms, be diagnosed as SUI according to gynaecological check-up, various auxiliary examination such as urinary dynamics inspection and urine mat test, and eliminate diagnosed unstable bladder, filling urinary incontinence, and urinary infection situation; etc. POP group:patients without stress incontinence symptoms, auxiliary examination both confirmed without incontinence, gynaecological check-up are moderate and above vaginal front wall surfaces.The control group:no incontinence symptoms, gynaecological check-up without pelvic organ prolapse. Postoperative histopathologic examination is proven to be an endometriosis etc estrogen related diseases.Based site:all take 12 points tissue of the anterior vaginal wall, 0.5cm×0.5cm×0.5cm. All selected patients didn't take estrogen drugs for six months before operation. SUI group using TVT-O, TVT-S operation or vaginal front wall suture, POP group were treated by former strengthening POP or vaginal front wall suture group, uterine full cut method (Yin type or abdominal). The sample inner-layer determination, the three groups of patients in the age, production times, body mass index, etc non-significance difference (P>0.05), all have comparability. Preoperative patients were informed agreement signed.2 MethodBy immunohistochemical S-P method, in strict accordance with the kit manual method dyeing, detection three groups of vaginal wall organization MMP-12, TIMP-1, Elastin expression, and at the same time use RT-PCR method detection three groups of patients vaginal wall tissue of Elastin expression at the molecular level.3 Image and results analysis3.1 Immunohistochemical methodIn a microscope, in vaginal wall submucosal connective tissue, the place where the indexes expressed occurred pale to tan yellow particles as positive cells. MMP-12 and TIMP-1 are mainly engaged in cytoplasm, Elastin is mainly located in the stroma. Comprehensive observe each expression, randomly selected five representative meaning of magnification vision (10 by 40) from which slice, adopts electronic computer image analyzer analy and calculate the each positive immunohistochemical expression of average gray values, between dyeing strength and average gray values become direct ratio.3.2 RT-PCR methodAfter electrophoresis, photographic record, analy the average gary values with geneβ-actin prescriptions as a benchmark.4 Statistic analysisentry all data with Excel 2003, and establish database, each group of grey value are with mean±standard(x±s)deviation said. SPSS 16.0 packages were being applied to statistically analyzed. Using single factor analysis of variance comparison between three groups of data, with LSD-t-test for binary comparison between groups, Pearson correlation analysis comparing correlation between each index. Various inspection methods are toα=0.05 for inspection standard.Results1 Immunohistochemical method 1.1 MMP-12 expression in each vaginal wall organizationMMP-12 were expressed in three groups of vaginal wall submucosal connective tissue plasminogen. In SUI group and POP group expressions are stronger than the control group, and the intensity between the SUI group and POP group of expression has not seen the obvious difference. SUI group, POP ergometry respectively comparative differences are statistically significant (P<0.05). Be compared between SUI group and POP group, differences was not statistically significant (P>0.05).1.2 TIMP-1 expression in each vaginal wall organizationTIMP-1 were expressed in three groups of vaginal wall submucosal connective tissue plasminogen. In SUI group and POP group expressions are weaker than the control group, and the intensity between the SUI group and POP group of expression has not seen the obvious difference. SUI group, POP ergometry respectively comparative differences are statistically significant (P<0.05). Be compared between SUI group and POP group, differences was not statistically significant (P>0.05).1.3 Elastin expression in each vaginal wall organizationElastin were expressed in three groups of vaginal wall submucosal connective tissue plasminogen. In SUI group and POP group expressions are weaker than the control group, and the intensity between the SUI group and POP group of expression has not seen the obvious difference. SUI group, POP ergometry respectively comparative differences are statistically significant (P<0.05). Be compared between SUI group and POP group, differences was not statistically significant (P> 0.05).1.4 the correlation analysis of MMP-12,TIMP-1, Elastin in SUI groupSUI group of vaginal wall organization with MMP-12 TIMP-1 express mean grey value of the relevant analysis, the result shows:a significant negative correlation between (r=-0.999, P<0.05). SUI group of vaginal wall organization MMP-12 and Elastin express mean grey value of the relevant analysis, the result shows:a significant negative correlation between (r=-0.614, P<0.05). TIMP-1 and Elastin expression is a significant positive correlation (r=0.628, P<0.05). 2 RT-PCR methodCalculation Elastin mRNA expression of vaginal wall 3 groups, in SUI group and POP group expression is weaker than the control group. And between the SUI group and POP group has not seen the obvious difference. SUI group, the POP group compared with controls are statistically significant difference (P<0.05), SUI group and POP group between comparative differences was not statistically significant (P>0.05).ConclusionMMP-12,TIMP-1 may participate the occurence of SUI by regulate content of Elastin. |
PDF Full Text Request