本文選題：壓力性 + 尿失禁　； 參考：《鄭州大學(xué)》2014年碩士論文

【摘要】：壓力性尿失禁(Stress Urinary Incontinence，SUI)被認(rèn)為是尿道對(duì)控尿起“開關(guān)-閥門”失控引起的功能障礙性疾病，多數(shù)患者由于分娩、創(chuàng)傷等引起肛提肌損傷、盆底筋膜和韌帶松弛、泌尿道解剖位置改變所致。SUI是一種多因素參與的復(fù)雜性疾病，往往伴隨女性盆腔器官脫垂（pelvic organ prolapse，POP）。其特點(diǎn)是平時(shí)無(wú)漏尿，如咳嗽、大笑、重體力勞動(dòng)等腹壓增加時(shí)，尿液不自主地自尿道流出，且癥狀嚴(yán)重程度隨年齡的增長(zhǎng)而加重，在美國(guó)60歲及以上38%婦女及17%男性均患有尿失禁。因此，國(guó)際控尿協(xié)會(huì)將SUI定義為腹壓突然增加時(shí)導(dǎo)致尿液失控不自主流出，其病因非逼尿肌收縮壓引起而是盆底解剖結(jié)構(gòu)出現(xiàn)異常或缺損致壓力傳導(dǎo)不均。有人將該病形象的稱為“社交癌”。雖然壓力性尿失禁不像心血管、腫瘤等疾病嚴(yán)重危及患者的生命健康，卻也引起了社會(huì)、經(jīng)濟(jì)和衛(wèi)生等不同程度的影響；其嚴(yán)重程度越來(lái)越受到廣大婦女的關(guān)注。隨著“吊床假說(shuō)”和“盆底整體理論”的提出，人們逐漸意識(shí)到僅單純檢測(cè)尿道壓、尿道長(zhǎng)度及膀胱尿道角等解剖改變的傳統(tǒng)方法已經(jīng)無(wú)法全面合理解釋SUI的發(fā)病機(jī)制，這就促使研究人員從分子生物學(xué)水平深入研究盆底支持組織的解剖結(jié)構(gòu)及功能。眾多研究表明，女性SUI患者盆底支持結(jié)構(gòu)中膠原及彈性蛋白含量顯著減少，引起支持組織彈性減弱，出現(xiàn)盆底松弛。而轉(zhuǎn)化生長(zhǎng)因子-β1（TGF-β1）作為最重要的細(xì)胞因子，既與膠原和彈性蛋白的合成及降解直接相關(guān)，又可改變基質(zhì)降解酶及其抑制因子的量及活性，在分娩、手術(shù)等造成盆底支持組織損傷修復(fù)過(guò)程中發(fā)揮關(guān)鍵性作用。結(jié)締組織生長(zhǎng)因子（CTGF）可刺激成纖維細(xì)胞增殖和分泌膠原，在纖維化性疾病、創(chuàng)傷后瘢痕形成等方面有較多的研究，然而在盆底障礙性疾病的研究國(guó)內(nèi)外少有報(bào)道。而Smad3蛋白是細(xì)胞內(nèi)TGF-β信號(hào)轉(zhuǎn)導(dǎo)分子，對(duì)細(xì)胞功能起著雙向調(diào)控作用，參與控制創(chuàng)面組織的愈合。 目的 本研究通過(guò)采用免疫組化和聚合酶-逆轉(zhuǎn)錄（RT-PCR）方法檢測(cè)TGF-β1、CTGF及Smad3三者在女性SUI患者陰道壁組織中的表達(dá)，分析三者之間的相關(guān)性，探討他們?cè)赟UI發(fā)生機(jī)制中的作用，期望為該病的預(yù)防、治療提供理論依據(jù)。 材料和方法 1.材料 選取2012年6月-2013年4月在本院婦產(chǎn)科收治的的60例中、重度壓力性尿失禁患者為實(shí)驗(yàn)組，包括25例單純性SUI，35例SUI合并POP，手術(shù)方式為經(jīng)陰道無(wú)張力尿道中段懸吊術(shù)（trans-obturator vaginal tape，TVT-O）、陰道前后壁修補(bǔ)術(shù)，陰道前壁懸吊術(shù)。同期選取20例宮頸上皮內(nèi)瘤變患者均不伴SUI為對(duì)照組，手術(shù)方式為腹式或陰式子宮切除術(shù)。3組在年齡、產(chǎn)次及體重指數(shù)等方面差異無(wú)統(tǒng)計(jì)學(xué)意義（P0.05），具有較好的可比性。所有病例均無(wú)類風(fēng)濕性關(guān)節(jié)炎、甲狀旁腺功能亢進(jìn)、肺氣腫和肝纖維化等影響膠原代謝的疾病，術(shù)前3個(gè)月內(nèi)未曾使用過(guò)性激素類藥物，未患功能性卵巢腫瘤，無(wú)泌尿生殖道感染，無(wú)既往盆底手術(shù)史。SUI診斷依據(jù)為患者的病史、婦科檢查、壓力試驗(yàn)、指壓試驗(yàn)、尿墊實(shí)驗(yàn)及尿動(dòng)力學(xué)檢查結(jié)果。SUI合并POP診斷依據(jù)是除具有壓力性尿失禁的診斷標(biāo)準(zhǔn)外，婦科檢查均發(fā)現(xiàn)合并有POP-Q分類法Ⅱ度及Ⅱ度以上的陰道前壁膨出。實(shí)驗(yàn)組于陰道前壁12點(diǎn)或膀胱宮頸韌帶周圍取兩塊陰道前壁組織各約0.8cm×0.5cm×0.2cm大小。對(duì)照組取自子宮切除后陰道殘端陰道壁組織。 2.實(shí)驗(yàn)方法 采用免疫組化（SP）法檢測(cè)SUI組、SUI+POP組和對(duì)照組患者陰道前壁組織中TGF-β1、CTGF和Smad3蛋白的表達(dá)情況。采用逆轉(zhuǎn)錄-聚合酶鏈?zhǔn)椒磻?yīng) （RT-PCR)技術(shù)檢測(cè)三組TGF-β1、CTGF和Smad3mRNA水平。 3.統(tǒng)計(jì)學(xué)分析 采用SPSS17.0統(tǒng)計(jì)軟件對(duì)數(shù)據(jù)處理。多組定性資料的比較采用獨(dú)立的多組二分類，兩兩比較采用四格表卡方檢驗(yàn)，檢驗(yàn)水準(zhǔn)取α/3=0.0167；多組定量資料的比較采用單因素方差分析，組間比較采用LSD-t法。兩連續(xù)變量相關(guān)分析采用Pearson積矩相關(guān)。檢驗(yàn)水準(zhǔn)α=0.05。 結(jié)果 1．免疫組化染色結(jié)果 SUI組、SUI+POP組和對(duì)照組3組患者陰道壁組織中均可見(jiàn)TGF-β1蛋白、CTGF蛋白和Smad3蛋白的表達(dá)。TGF-β1和CTGF蛋白主要表達(dá)于胞漿，呈現(xiàn)淡黃色至棕黃褐色的細(xì)胞為陽(yáng)性細(xì)胞；Smad3蛋白主要表達(dá)于細(xì)胞核，呈現(xiàn)淡黃色至棕黃褐色顆粒的細(xì)胞為陽(yáng)性細(xì)胞。 1) TGF-β1蛋白在3組陰道壁中陽(yáng)性表達(dá)率分別為28%、17.14%、95%，3組之間差異有顯著性（χ2=34.327，P0.01）；SUI組和SUI+POP組比較，差異無(wú)顯著性（P0.05）。 2) CTGF蛋白在3組陰道壁中陽(yáng)性表達(dá)率分別為16%、20%、95%，3組之間差異有顯著性（χ2=37.717，P0.01）；SUI組和SUI+POP組比較，差異無(wú)顯著性（P0.05）。 3) Smad3蛋白在3組陰道壁中陽(yáng)性表達(dá)率分別為24%、14.29%、90%，3組之間差異有顯著性（χ2=33.933，P0.01）；SUI組和SUI+POP組比較，差異無(wú)顯著性（P0.05）。 2. RT-PCR結(jié)果半定量測(cè)定結(jié)果顯示：TGF-β1mRNA、CTGFmRNA及Smad3mRNA在3組中均為陽(yáng)性表達(dá)。 1) TGF-β1mRNA在SUI組、SUI+POP組和對(duì)照組陰道壁組織中表達(dá)量分別為（0.294±0.013）、（0.293±0.019）和（0.610±0.052），SUI組、SUI+POP組與對(duì)照組比較差異均有顯著性（P0.01）；而SUI組與SUI+POP組比較差異無(wú)顯著性（P0.05）。 2) CTGF mRNA在SUI組、SUI+POP組和對(duì)照組陰道壁組織中表達(dá)量分別為（0.085±0.012）、（0.092±0.011）和（0.272±0.029），SUI組、SUI+POP組與對(duì)照組比較差異均有顯著性（P0.01）；而SUI組與SUI+POP組比較差異無(wú)顯著性（P0.05）。 3) Smad3mRNA在SUI組、SUI+POP組和對(duì)照組陰道壁組織中表達(dá)量分別為（0.091±0.011）、（0.088±0.012）和（0.272±0.029），SUI組、SUI+POP組與對(duì)照組比較差異均有顯著性（P0.01）；而SUI組與SUI+POP組比較差異無(wú)顯著性（P0.05）。 3. SUI患者組織中TGF-β1和CTGF的關(guān)聯(lián)性分析 相關(guān)分析顯示：SUI組陰道壁組織中TGF-β1與CTGF呈正相關(guān)（r=0.959，P0.05），TGF-β1與Smad3的表達(dá)呈正相關(guān)（r=0.944，P0.05），CTGF與Smad3的表達(dá)呈正相關(guān)（r=0.965，P0.05）。 結(jié)論 TGF-β1、CTGF及Smad3三者表達(dá)均降低，提示其可能參與了壓力性尿失禁的發(fā)生。
[Abstract]:Stress urinary incontinence (Stress Urinary Incontinence, SUI) is considered to be a dysfunctional disease caused by the urethral "switch valve" out of control of the urinary tract. Most patients are caused by labor, trauma, and other injuries of the muscle of the anus, the pelvic floor fascia and ligamentum laxity, and the change of the urinary anatomic position caused by.SUI is a complex disease involved in multiple factors. The disease is often accompanied by pelvic organ prolapse (POP). It is characterized by urinary incontinence when no leakage of urine, such as coughing, laughter, and heavy manual labor, increases with age. Therefore, 38% women and 17% men in the United States have urinary incontinence. The International Association for control of the urine (SUI) defines an involuntary outflow of runaway urine resulting from a sudden increase in abdominal pressure. The cause of the cause is not detrusor contraction pressure, but an abnormal or defective stress conduction in the pelvic anatomy. Some people call the image "social cancer". Although stress urinary incontinence is not as serious as cardiovascular and tumor diseases Endanger the patient's life and health, but it also causes social, economic and health effects, and its severity is becoming more and more popular with women. With the "hammock hypothesis" and "pelvic floor holistic theory", people are gradually aware of the anatomic changes of urethral pressure, urethral length and bladder urethra angle. The traditional methods have been unable to explain the pathogenesis of SUI in a comprehensive and rational way. This has prompted researchers to study the anatomical structure and function of pelvic floor support from the molecular biology level. Many studies have shown that the content of collagen and elastin in the pelvic floor support structure of female SUI patients is significantly reduced, and the elasticity of support tissue is weakened and the basin is present. The growth factor - beta 1 (TGF- beta 1), as the most important cytokine, is not only directly related to the synthesis and degradation of collagen and elastin, but also changes the amount and activity of the matrix degrading enzyme and its inhibitory factors. It plays a key role in the repair of pelvic floor support injury in childbirth and surgery. Factor (CTGF) stimulates the proliferation and secretion of collagen in fibroblasts. There are many studies in the aspects of fibrotic diseases and posttraumatic scar formation. However, few reports have been made in the study of pelvic floor disorders at home and abroad. The Smad3 protein is a intracellular TGF- beta signal transduction molecule, which plays a bidirectional regulatory role in cell function and participates in the control of the wound. The healing of the tissue.
objective
In this study, the expression of TGF- beta 1, CTGF and Smad3 in the vaginal wall tissue of female SUI patients was detected by immunohistochemistry and polymerase chain reverse transcription (RT-PCR) method, and the correlation between the three groups was analyzed and their role in the pathogenesis of SUI was discussed. The theoretical basis was provided for the prevention and treatment of the disease.
Materials and methods
1. material
Among the 60 cases of severe stress urinary incontinence admitted in June 2012 -2013 and April in our department of Obstetrics and Gynecology, severe stress incontinence was included in the experimental group, including 25 cases of simple SUI, 35 cases of SUI combined with POP, and the operation method was transvaginal tension-free urethral suspension (trans-obturator vaginal tape, TVT-O), vaginal wall repair and anterior vaginal wall suspension. 20 cases of cervical intraepithelial neoplasia were selected without SUI as the control group. The surgical methods were abdominal or vaginal hysterectomy in group.3 with no statistically significant difference in age, birth order and body mass index (P0.05), and had better comparability. All cases had no rheumatoid arthrosis, hyperparathyroidism, emphysema and liver fibrosis. No sex hormone drugs have been used in 3 months before operation, no functional ovarian tumors, no urogenital tract infection, no history of previous pelvic floor surgery.SUI diagnosis is based on patients' history, gynecologic examination, stress test, pressure test, urinary cushion test and urodynamic examination results of.SUI combined with POP diagnosis basis Except for the diagnostic criteria for stress urinary incontinence, the gynecologic examination found the anterior vaginal wall bulge with POP-Q classification II and more than 2 degrees. The experimental group took about 0.8cm x 0.5cm x 0.2cm in the anterior wall of the vagina at 12 points of the anterior vaginal wall or around the cervix ligament of the bladder. The vaginal wall of the vaginal stump after hysterectomy was taken from the group. Organization.
2. experimental method
The expression of TGF- beta 1, CTGF and Smad3 in the anterior vaginal wall of group SUI, SUI+POP group and control group was detected by immunohistochemistry (SP). The reverse transcription polymerase chain reaction was used.
(RT-PCR) techniques were used to detect the levels of TGF- beta 1, CTGF and Smad3mRNA in the three groups.
3. statistical analysis
SPSS17.0 statistical software was used for data processing. Multiple groups of qualitative data were compared by independent multiple groups of two classifications, 22 compared with four lattice chi square tests and test level for alpha /3=0.0167; multiple quantitative data were compared by single factor analysis of variance and LSD-t method was used for comparison between groups. Two continuous variable correlation analysis adopted Pearson moment analysis. Correlation. Test level alpha =0.05.
Result
1. immunohistochemical staining results
TGF- beta 1 protein was found in the vaginal wall of group SUI, group SUI+POP and control group. The expression of.TGF- beta 1 and CTGF protein expressed mainly in the cytoplasm, and the positive cells were yellowish to brown brown cells, and Smad3 protein was mainly expressed in the nucleus, and the cells were yellowish to brown brown granules. Positive cells.
1) the positive expression rate of TGF- beta 1 protein in the 3 groups of vaginal walls were 28%, 17.14%, 95%, respectively. The difference between the 3 groups was significant (x 2=34.327, P0.01), and there was no significant difference between the SUI and SUI+POP groups (P0.05).
2) the positive expression rate of CTGF protein in the 3 groups of vaginal walls was 16%, 20%, 95%, and the difference between the 3 groups was significant (x 2=37.717, P0.01), and there was no significant difference between the SUI group and the SUI+POP group (P0.05).
3) the positive expression rate of Smad3 protein in the 3 groups of vaginal walls was 24%, 14.29%, 90%, and the difference between the 3 groups was significant (x 2=33.933, P0.01), and there was no significant difference between the SUI group and the SUI+POP group (P0.05).
2. RT-PCR results semi quantitative analysis showed that TGF- beta 1mRNA, CTGFmRNA and Smad3mRNA were all positive in the 3 groups.
1) the expression of TGF- beta 1mRNA in SUI group, SUI+POP group and control group was (0.294 + 0.013), (0.293 + 0.019) and (0.610 + 0.052), SUI group and SUI+POP group were significantly different from those of control group (P0.01), but there was no significant difference between group SUI and SUI+POP group (P0.05).
2) the expression of CTGF mRNA in SUI group, SUI+POP group and control group was (0.085 + 0.012), (0.092 + 0.011) and (0.272 + 0.029), SUI group and SUI+POP group were significantly different from those of control group (P0.01), but there was no significant difference between group SUI and SUI+POP group (P0.05).
3) the expression of Smad3mRNA in SUI group, SUI+POP group and control group was (0.091 + 0.011), (0.088 + 0.012) and (0.272 + 0.029), SUI group, SUI+POP group was significantly different from that of control group (P0.01), but there was no significant difference between group SUI and SUI+POP group (P0.05).
Correlation analysis between TGF- beta 1 and CTGF in 3. SUI patients
The correlation analysis showed that the TGF- beta 1 in the vaginal wall of SUI group was positively correlated with CTGF (r=0.959, P0.05), and TGF- beta 1 was positively correlated with the expression of Smad3 (r=0.944, P0.05), and there was a positive correlation between the expression of CTGF and Smad3 (r=0.965,).
conclusion
The expression of TGF- beta 1, CTGF and Smad3 decreased, suggesting that it may be involved in the occurrence of stress urinary incontinence (three).

【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R711.59

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 孫智晶,郎景和,朱蘭,陳杰;膠原狀態(tài)與壓力性尿失禁的病因?qū)W研究[J];中國(guó)婦產(chǎn)科臨床雜志;2004年01期

2 郭鳳琴;洪莉;;盆底器官脫垂患者恥骨宮頸筋膜中TGF-β_1和CTGF的表達(dá)[J];武漢大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年01期

3 蘇sセ，

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