發(fā)布時間：2018-05-27 10:45

  本文選題：壓力性尿失禁 + AQP2��； 參考：《浙江大學(xué)》2014年碩士論文

【摘要】：背景和目的： 壓力性尿失禁(stress urinary incontinence, SUI)是中老年婦女常見慢性病,是指在腹壓增加時,出現(xiàn)不自主的尿液自尿道外口溢出。SUI的病因很復(fù)雜,年齡是其發(fā)病的主要誘因之一,但是發(fā)病機制迄今未明,最經(jīng)典的“整體理論”和“吊床假說”均強調(diào)尿道支撐結(jié)構(gòu)薄弱導(dǎo)致SUI的發(fā)生。尿道支撐結(jié)構(gòu)由大量細胞外基質(zhì)(extracellular matrix, ECM)和少量成纖維細胞等組成。有研究表明ECM重構(gòu)障礙是SUI發(fā)病的關(guān)鍵因素,而雌激素直接參與了ECM的重構(gòu)過程。水通道蛋白(Aquaporins AQPs)是選擇性高效轉(zhuǎn)運水分子的通道,同時還廣泛參與人體細胞的遷移運動,與多種組織的損傷修復(fù)有關(guān),而且有研究發(fā)現(xiàn)在水通道蛋白2(Aquaporin2,AQP2)的基因上存在雌激素的反應(yīng)元件,證實了雌激素能直接調(diào)控AQP2的基因轉(zhuǎn)錄而調(diào)節(jié)其在子宮內(nèi)膜細胞上的表達,從而促進內(nèi)膜細胞的遷移、入侵、粘附及增殖。本研究擬通過原代培養(yǎng)獲得女性尿道支撐組織成纖維細胞,采用免疫熒光技術(shù)證實AQP2在尿道支撐組織成纖維細胞上的表達,采用Western blot法觀測絕經(jīng)前后SUI患者與非SUI患者尿道支撐組織成纖維細胞上AQP2的表達差異,探討AQP2在SUI發(fā)生發(fā)展中的作用；同時,通過測定血清雌二醇(estradiol,E2)水平,探討尿道支撐組織成纖維細胞上AQP2的表達水平與血清E2水平的相關(guān)性。 方法： 收集浙江大學(xué)醫(yī)學(xué)院附屬婦產(chǎn)科醫(yī)院2013年1月至2014年3月收治的18例SUI患者(SUI組)和18例非壓力性尿失禁婦女(對照組)的陰道前壁組織(即尿道支撐組織),每組再分為絕經(jīng)前后兩組各9例,所有組織標本來源于SUI患者行抗尿失禁手術(shù)時和非SUI患者因尿道旁腺囊腫、尿道憩室、尿道附近陰道壁囊腫等疾病手術(shù)時,取材于尿道附近約0.5×1cm2的陰道前壁全層組織。取組織塊進行成纖維細胞原代培養(yǎng),并予免疫組化鑒定。采用免疫熒光法測定成纖維細胞上AQPs各亞型的表達,采用Western blot法檢測成纖維細胞上AQP2蛋白表達水平,比較絕經(jīng)前后SUI組和非SUI組之間尿道支撐組織成纖維細胞上AQP2蛋白表達水平的差異。 所有36例入選對象均在外科手術(shù)前一天早晨7時空腹采靜脈血2ml,采用電化學(xué)發(fā)光免疫檢測(ECLIA)法檢測血清E2水平,采用Pearson相關(guān)性分析法探討尿道支撐組織成纖維細胞上AQP2蛋白表達水平與血清E2水平之間的相關(guān)性。 結(jié)果： 1.免疫熒光技術(shù)和Western blot法均發(fā)現(xiàn)女性尿道支撐組織成纖維細胞上有AQP2的表達； 2.經(jīng)t檢驗,絕經(jīng)前后SUI組和對照組的年齡、分娩次數(shù)、體重指數(shù)以及絕經(jīng)后SUI組和對照組的絕經(jīng)年限差異均無統(tǒng)計學(xué)意義；絕經(jīng)前血清E2水平SUI組(311.43±93.63nmol/L)顯著低于對照組(530.30±232.21nmol/L)(P0.05).絕經(jīng)后血清E2水平SUI組為39.91±16.76nmol/L,對照組為31.30±9.95nmol/L兩組之間差異無統(tǒng)計學(xué)意義(P0.05)；經(jīng)相關(guān)性分析,血清E2水平與所有標本尿道支撐組織成纖維細胞(r=—0.537,P0.05)及總的SUI組標本尿道支撐組織成纖維細胞(r=—0.860,P0.05)上AQP2的表達水平均存在顯著的負相關(guān),而與總的對照組標本尿道支撐組織成纖維細胞上AQP2的表達水平無顯著相關(guān)(P0.05)。 3.經(jīng)Western blot法檢測,絕經(jīng)前后SUI組及絕經(jīng)前后對照組尿道支撐組織成纖維細胞上均有AQP2的表達,總的SUI組和總的對照組尿道支撐組織成纖維細胞上AQP2的蛋白表達水平分別為26.29±2.53和25.32±3.80,t檢驗顯示兩組間差異無統(tǒng)計學(xué)意義(P0.05)；總的絕經(jīng)前組和總的絕經(jīng)后組尿道支撐組織成纖維細胞上AQP2的蛋白表達水平分別為23.25±2.40和28.36±1.31,經(jīng)t檢驗,總的絕經(jīng)后組尿道支撐組織成纖維細胞上AQP2的表達明顯升高(P0.05)絕經(jīng)前SUI組和絕經(jīng)后SUI組尿道支撐組織成纖維細胞上AQP2的蛋白表達水平分別為24.03±0.98和28.54±1.11,絕經(jīng)前對照組和絕經(jīng)后對照組尿道支撐組織成纖維細胞上AQP2的蛋白表達水平分別為22.46±3.15和28.19+1.54,經(jīng)t檢驗,絕經(jīng)前SUI組與絕經(jīng)前對照組、絕經(jīng)后SUI組與絕經(jīng)后對照組上的表達差異均無統(tǒng)計學(xué)意義(P均0.05)；絕經(jīng)后SUI組尿道支撐組織成纖維細胞上AQP2的表達較絕經(jīng)前SUI組上的表達明顯升高,兩組間的比較差異有統(tǒng)計學(xué)意義(P0.05)；絕經(jīng)后對照組尿道支撐組織成纖維細胞上AQP2的表達較絕經(jīng)前對照組上的表達亦明顯升高,兩組間的比較差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論： 1.女性尿道支撐組織成纖維細胞上存在AQP2的表達； 2.AQP2的表達與雌激素水平密切相關(guān),絕經(jīng)后SUI組尿道支撐組織成纖維細胞上AQP2的表達較絕經(jīng)前SUI組上的表達明顯升高,可能是絕經(jīng)后尿道支撐組織成纖維細胞上低雌激素水平上調(diào)AQP2的表達所致。
[Abstract]:Background and Purpose :

Aquaporins ( AQPs ) is a key factor for selective high - efficiency transport of water molecules , and it has been found that estrogen can regulate the expression of AQP2 directly and regulate the expression of AQP2 on endometrial cells .
At the same time , the level of serum estradiol ( E2 ) was measured , and the correlation between AQP2 expression level and serum E2 level was investigated .

Method :

The expression of AQP2 protein in fibroblasts from January 2013 to March 2014 was collected from January 2013 to March 2014 in the Affiliated Hospital of Zhejiang University Medical College from January 2013 to March 2014 .

The serum levels of E2 were measured by electrochemical luminescence immunoassay ( ECLIA ) and Pearson correlation analysis was used to investigate the correlation between AQP2 protein expression level and serum E2 level .

Results :

1 . The expression of AQP2 was found in the fibroblasts of female urethra by immunofluorescence technique and Western blot .


2 . After t test , there was no significant difference between the age , the number of deliveries , the body mass index and the menopausal period in the control group before and after menopause .
The serum level of E2 in the postmenopausal serum was significantly lower than that of the control group ( 530.30 鹵 232.21nmol / L ) ( P0.05 ) . The serum level of E2 in postmenopausal serum was 39.91 鹵 16.76nmol / L and 31.30 鹵 9.95nmol / L in the control group ( P0.05 ) .
The level of serum E2 was negatively correlated with the expression level of AQP2 in all specimens of urethral support tissue fibroblasts ( r = - 0.537 , P0.05 ) , and the expression level of AQP2 was not significantly correlated with the total control group ( P0.05 ) .

3 . The expression level of AQP2 was detected by Western blot , and the expression level of AQP2 was 26.29 鹵 2.53 and 25.32 鹵 3.80 respectively , and there was no significant difference between the two groups ( P0.05 ) .
The expression levels of AQP2 were 23.25 鹵 2.40 and 28.36 鹵 1.31 , respectively , and the protein expression levels of AQP2 were 24.03 鹵 0.98 and 28.54 鹵 1.11 respectively .
The expression of AQP2 was significantly higher than that in premenopausal women , but there was significant difference between the two groups ( P0.05 ) .
Compared with the control group , the expression of AQP2 in the control group was significantly higher than that in the control group ( P0.05 ) .

Conclusion :

1 . The expression of AQP2 was found in the fibroblasts of female urethra .


2 . The expression of AQP2 is closely related to the level of estrogen , and the expression of AQP2 is obviously higher than that in premenopausal women , which may be the result of the up - regulation of AQP2 by low estrogen level in the post - menopausal urethral support tissue fibroblasts .
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R711.59

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