本文選題：壓力性尿失禁 + 免疫組化��； 參考：《暨南大學(xué)》2012年碩士論文

【摘要】：研究目的：本研究通過建立SUI大鼠模型，檢測SD雌鼠尿流動力學(xué)變化，觀察尿道及其周圍組織學(xué)形態(tài)改變。應(yīng)用免疫組化和圖像分析方法，分析HUMSCs治療前后desmin、MHC及PGP9.5三種因子在尿道中的表達(dá)情況，分析間充質(zhì)干細(xì)胞治療壓力性尿失禁的效果和可能的機制，探討兩種不同方式干細(xì)胞移植后的效果，以指導(dǎo)臨床。 材料和方法：1.1選用SD（清潔級）雌鼠模擬人類妊娠、難產(chǎn)產(chǎn)傷、雙側(cè)卵巢去勢后低雌激素建立SUI模型。測定實驗動物的最大膀胱容量(MBC)、漏尿點壓(LPP),噴嚏實驗陽性率和組織學(xué)形態(tài)檢驗?zāi)Ｐ偷某晒εc否。通過HE染色、Masson特殊染色、Mallory特殊染色、免疫組化方式對大鼠尿道括約肌的顯微解剖進(jìn)行研究。 2.2將建模成功好的20只大鼠分為三組：靜脈注射組（A組，6只），尿道局部注射組(B組，8只)、對照組（不注射任何物質(zhì)C組，6只)，D組正常組（6只）將獲得MSCs分別以5*106個細(xì)胞/只通過尾靜脈和尿道局部注射方式移植于SUI大鼠模型體內(nèi)，移植4周及6周后分別用BL-410生物機能實驗系統(tǒng)檢測尿流動力學(xué)指標(biāo)和噴嚏實驗陽性率，以評估尿流動力學(xué)變化。 2.3注射細(xì)胞6周后處死所有大鼠，進(jìn)行尿道及其周圍組織取材，通過組織形態(tài)學(xué)和圖像分析法觀察尿道及其周圍組織變化，定量評價desmin、MHC、PGP9.5含量和計算橫紋肌占整個尿道陽性面積比及尿道括約肌占整個尿道的陽性面積比。 結(jié)果：1.正常SD雌鼠尿道及其周圍組織形態(tài)學(xué)觀察尿道括約肌在整個尿道分布不均，在中下段開始增多，其中橫紋肌呈環(huán)形分布。模擬人類妊娠、難產(chǎn)產(chǎn)傷、去勢后低雌激素方式能建立SUI模型SD雌鼠:尿道括約肌肌層減少，排列松散，混亂，部分肌肉有斷裂，形成空泡等現(xiàn)象，，肌相關(guān)蛋白desmin及MHC染色下降，甚至出現(xiàn)丟失現(xiàn)象。 2.LPP是反應(yīng)尿控功能最重要的指標(biāo)，其中LPP升高提示尿控功能得到部分恢復(fù)或完全恢復(fù)，本研究顯示HUMSCs移植A、B兩組LPP、MBC均增加，HUMSCs移植后4周LPP值A(chǔ)組、B組、C組、D組分別為：18.15±1.21mmHg，18.79±1.48mmHg,13.55±1.43mmHg，22.83±2.17mmHg，6周后LPP分別為18.86±1.14mmHg,19.40±0.85mmHg，11.82±0.91mmHg,24.36±3.26mmHg.組間兩兩比較有統(tǒng)計學(xué)差異(P0.01)，其中C組LPP最小，A組、B組及D組兩兩比較有統(tǒng)計學(xué)意義（P0.01），但是A與B組比較無統(tǒng)計學(xué)意義(P0.05)，HUMSCs治療后4周與6周各組間LPP無統(tǒng)計學(xué)意義(P0.05)。 3．HUMSCs移植治療后可以使尿道括約肌肌層得到明顯的增厚，形態(tài)及功能得到明顯的改善。圖像分析顯示治療4周、6周后尿道橫紋肌及括約肌陽性面積比較未治療都有明顯的增加，PGP9.5數(shù)目也較陽性對照組明顯增多,A組、B組、C組、D組橫紋肌陽性面積比(%)分別為(15.3±3.77)%,(15.2±4.79)%,(9.55±2.06)%,(16.49±2.4)%,；尿道括約肌陽性面積比(%)分別為(35.14±5.38)%,(36.88±3.93)%,(22.46±3.52)%,(37.9±1.80)%, PGP個數(shù)分別為7.75±1.26，8.14±1.68，5.2+2.05，10.25+1.89，上述三種因子的檢測組間兩兩比較均有統(tǒng)計學(xué)差異(P0.01)，其中C組橫紋肌陽性面積最小，與其他三組相比差異有統(tǒng)計學(xué)意義(P0.01)，但A組、B組及D組兩兩比較無統(tǒng)計學(xué)意義(P0.05)； 結(jié)論:1.本部分研究通過妊娠、分娩陰道擴張及雙側(cè)卵巢切除去勢的方法，成功建立了SUI的動物模型，為后續(xù)實驗奠定了基礎(chǔ)。 2.尿道括約肌損傷和神經(jīng)纖維的減少導(dǎo)致SUI出現(xiàn)，提示尿道結(jié)構(gòu)和功能的改變是引發(fā)SUI的直接原因。 3.HUMSCs移植到SUI模型SD雌鼠可以增加肌相關(guān)蛋白desmin、MHC的含量和神經(jīng)軸突標(biāo)記物PGP9.5數(shù)目。 4.局部注射治療和靜脈注射治療的兩種不同方式療效無明顯的區(qū)別。 5.HUMSCs移植到SUI模型SD雌鼠可修復(fù)薄弱的尿道括約肌，改善尿控功能。
[Abstract]:Objective : To investigate the changes of urinary flow dynamics of SD female rats and to observe the changes of histological morphology around the urethra and its surrounding tissues by establishing a rat model . The effects of mesenchymal stem cells in the treatment of stress urinary incontinence and possible mechanisms were analyzed . The effects of mesenchymal stem cells on stress urinary incontinence were analyzed , and the effect of two different methods of stem cell transplantation was discussed .

Materials and Methods : 1.1 Female rats with SD ( clean grade ) were selected to simulate human pregnancy , which was difficult to produce and produce low estrogen after castration . The results showed that the maximal bladder capacity ( MBC ) , urine leakage point pressure ( lpp ) , the positive rate and the histological morphology of the experimental animals were successful or not . The microscopic anatomy of the urethral sphincter in rats was studied by HE staining , special staining of the Mallory , and immunohistochemical method .

2.2 All 20 rats with successful modeling were divided into three groups : IV group ( group A , 6 rats ) , local injection group ( group B , 8 rats ) , control group ( no substance C group , 6 rats ) , normal group D group ( 6 rats ) .

2 . After 6 weeks of injection , all the rats were sacrificed and the urethra and its surrounding tissues were taken , and the changes of the urethra and its surrounding tissues were observed by histomorphology and image analysis . The contents of Desin , MHC , PGP9.5 were quantitatively evaluated .

Results : 1 . The morphology of the urethra and its surrounding tissues in normal SD female rats showed that the urethral sphincter was not evenly distributed throughout the urethra , and the middle and lower segments began to grow .

There was significant difference between group A , group B , group C and D in group A , group B , group C , and D group ( P 0.01 ) , but there was no statistical difference between group A and group B ( P0.05 ) , but there was no statistical difference between group A and group B ( P0.05 ) .

3 . After the transplantation of HUMSCs , the thickness , shape and function of the urethral sphincter muscle were obviously improved . The results showed that the positive area of the striated muscle and sphincter was increased significantly after 4 weeks and 6 weeks . The positive area ratio of the striated muscle in group A , group B , group C and D was ( 15.3 鹵 3.77 ) % , ( 15.2 鹵 4.79 ) % , ( 9.55 鹵 2.06 ) % , ( 16.49 鹵 2.4 ) % , respectively .
The positive area ratio ( % ) of the urethral sphincter was ( 35.14 鹵 5.38 ) % , ( 36.88 鹵 3.93 ) % , ( 22.46 鹵 3.52 ) % , ( 37.9 鹵 1.80 ) % , respectively ( P < 0.01 ) .


Conclusion : 1 . In this part , this part studies the method of pregnancy , vaginal dilation and bilateral oophorectomy and lays a foundation for the follow - up experiment .

2 . The urethral sphincter injury and the decrease in nerve fibers lead to the occurrence of the occurrence of the disease , suggesting a change in the structure and function of the urethra is a direct cause of the cause of the disease .

3 . The transplantation of HUMSCs into the model SD female mice could increase the content of myoassociated proteins , and the content of MHC and the number of nerve axon markers PGP9.5 .

4 . There was no significant difference between the two different methods of local injection therapy and IV therapy .

5 . The transplantation of HUMSCs into the model SD female mice could repair the weak urethral sphincter and improve the urinary control function .
【學(xué)位授予單位】：暨南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉小璇;樊東升;張俊;鄭菊陽;馬鐵民;;比較不同方式移植的人間充質(zhì)干細(xì)胞在大鼠失神經(jīng)支配的骨骼肌的分布及對坐骨神經(jīng)損傷的影響[J];北京大學(xué)學(xué)報(醫(yī)學(xué)版);2008年02期

2 胡麗娟;杜廣輝;胡衛(wèi)峰;陳彪;章慧平;葉章群;;雌性大鼠尿道括約肌組織結(jié)構(gòu)的增齡變化[J];第三軍醫(yī)大學(xué)學(xué)報;2009年19期

3 呂璐璐;劉擁軍;許貞書;王彤;張浪輝;朱雄鵬;梁琳慧;王晗;陳志哲;韓忠朝;;臍帶源間充質(zhì)干細(xì)胞的分離和生物學(xué)性狀[J];福建醫(yī)科大學(xué)學(xué)報;2006年02期

4 羅新;高綠芬;陳翠平;謝靜燕;蔣學(xué)風(fēng);沈媛;;17β-雌二醇對骨髓間充質(zhì)干細(xì)胞增殖及成肌分化作用的研究[J];中國婦產(chǎn)科臨床雜志;2010年05期

5 鄭捷;方祖軍;丁強;;壓力性尿失禁與尿道周圍肌細(xì)胞凋亡的關(guān)系[J];上海交通大學(xué)學(xué)報(醫(yī)學(xué)版);2008年07期

6 楊艷;吳克明;;更年歸腎丸對雌性去勢大鼠性激素與下丘腦神經(jīng)遞質(zhì)調(diào)節(jié)作用的實驗研究[J];世界中西醫(yī)結(jié)合雜志;2007年08期

7 姜洋;金曉明;屠康;;平均陽性染色面積百分比法分析免疫組化結(jié)果初探[J];生物醫(yī)學(xué)工程學(xué)雜志;2007年03期

8 蔣芳;朱蘭;郎景和;姜雪鶯;;盆底功能障礙性疾病患者陰道前后壁中蛋白基因產(chǎn)物9.5和血管活性腸肽的表達(dá)研究[J];生殖醫(yī)學(xué)雜志;2008年06期

9 吳桂珠;鄭秀;江忠清;王金華;宋巖峰;;Induced Differentiation of Adipose-derived Stromal Cells into Myoblasts[J];Journal of Huazhong University of Science and Technology(Medical Sciences);2010年03期

10 王忠瓊;李昌平;杜光紅;鐘曉琳;陳霞;;多種細(xì)胞因子誘導(dǎo)骨髓間充質(zhì)干細(xì)胞向肝細(xì)胞的分化[J];中國組織工程研究與臨床康復(fù);2008年21期

相關(guān)博士學(xué)位論文 前2條

1 陳園;胰島素樣生長因子-Ⅰ對雌性大鼠尿道括約肌損傷修復(fù)的影響及其機制研究[D];華中科技大學(xué);2008年

2 吳桂珠;脂肪間充質(zhì)干細(xì)胞治療壓力性尿失禁的實驗研究[D];福建醫(yī)科大學(xué);2010年

相關(guān)碩士學(xué)位論文 前1條

1 姚潤斯;間充質(zhì)干細(xì)胞移植治療壓力性尿失禁大鼠的實驗研究[D];暨南大學(xué);2009年

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