妊娠和分娩對大鼠肛提肌細(xì)胞骨架的影響
本文選題:盆底功能障礙性疾病 + 妊娠; 參考:《河北醫(yī)科大學(xué)》2014年碩士論文
【摘要】:目的: 1研究妊娠和分娩對大鼠肛提肌細(xì)胞骨架蛋白α-肌動(dòng)蛋白(α-actin)、微管蛋白(tubulin)、聯(lián)絲蛋白(synemin)含量變化及mRNA表達(dá)的影響。 2探究妊娠和分娩與細(xì)胞骨架的關(guān)系。 3探討妊娠和分娩引起的肛提肌細(xì)胞骨架變化在女性盆底功能障礙性疾。╬elvic floor dysfunction,PFD)發(fā)病機(jī)制中的作用。 方法: 1研究對象:選用健康的Wistar育齡大鼠,,取20只未交配的雌鼠作為非妊娠組;再取60只孕鼠隨機(jī)分為妊娠晚期、產(chǎn)后6小時(shí)、產(chǎn)后6天3組,每組20只。適應(yīng)性喂養(yǎng)后各組均適時(shí)處死取肛提肌。(由河北醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物部供給) 2取材:由河北醫(yī)科大學(xué)實(shí)驗(yàn)動(dòng)物部供給的大鼠的種屬、窩別、年齡、體重和營養(yǎng)狀況與進(jìn)食量等方面無差異。所取標(biāo)本為肛提肌,大小約0.5cm×0.5cm×0.5cm,所取標(biāo)本經(jīng)10%中性福爾馬林溶液固定及液氮保存。 3應(yīng)用免疫組織化學(xué)過氧化物酶標(biāo)記鏈霉卵白素法(IHC SP法)和實(shí)時(shí)熒光定量PCR法(RT-PCR),分別檢測非妊娠組、妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組大鼠肛提肌細(xì)胞中α-肌動(dòng)蛋白、微管蛋白、聯(lián)絲蛋白的表達(dá)。 4用SPSS13.0統(tǒng)計(jì)軟件包,對所得數(shù)據(jù)進(jìn)行統(tǒng)計(jì)分析,以P0.05為有統(tǒng)計(jì)學(xué)意義。 結(jié)果: 1免疫組織化學(xué)結(jié)果:在非妊娠組、妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組大鼠肛提肌細(xì)胞中均檢測到α-肌動(dòng)蛋白(α-actin)、微管蛋白(tubulin)、聯(lián)絲蛋白(synemin)的表達(dá),α-肌動(dòng)蛋白在非妊娠組、妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組肛提肌細(xì)胞中表達(dá)水平的差異具有顯著統(tǒng)計(jì)學(xué)意義(P<0.05)。微管蛋白在非妊娠組、妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組肛提肌細(xì)胞中表達(dá)水平的差異具有顯著統(tǒng)計(jì)學(xué)意義(P<0.05)。聯(lián)絲蛋白在非妊娠組、妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組肛提肌細(xì)胞中表達(dá)水平的差異具有顯著統(tǒng)計(jì)學(xué)意義(P<0.05)。隨著妊娠分娩及產(chǎn)后恢復(fù)的過程,α-肌動(dòng)蛋白和聯(lián)絲蛋白表達(dá)量先減少后增加,微管蛋白的表達(dá)量先增加后減少,但是組間比較發(fā)現(xiàn),α-肌動(dòng)蛋白在非妊娠組和妊娠晚期組(P=0.084)中、微管蛋白在妊娠晚期組和產(chǎn)后6天組(P=0.836)中的表達(dá)無顯著差異性。其余組間比較均有顯著差異性。2熒光定量PCR結(jié)果:α-肌動(dòng)蛋白mRNA產(chǎn)后6小時(shí)組、產(chǎn)后6天組表達(dá)量(0.726±0.021,0.906±0.113)較非妊娠組(1.000±0.000)下降后上升,差異有統(tǒng)計(jì)學(xué)意義,P<0.05,妊娠晚期組mRNA表達(dá)量(0.975±0.010)下降,與非妊娠組差異無統(tǒng)計(jì)學(xué)意義,P>0.05;微管蛋白mRNA妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組表達(dá)量(1.652±0.085,5.905±0.13,3.059±0.177)較非妊娠組(1.000±0.000)先升高后下降,各組間差異均有統(tǒng)計(jì)學(xué)意義,P<0.05;聯(lián)絲蛋白mRNA妊娠晚期組、產(chǎn)后6小時(shí)組、產(chǎn)后6天組表達(dá)量(0.865±0.026,0.532±0.042,0.735±0.014)較非妊娠組(1.000±0.000)先下降后升高,各組間差異均有統(tǒng)計(jì)學(xué)意義,P<0.05。 結(jié)論: 1妊娠與分娩引起大鼠α-肌動(dòng)蛋白、聯(lián)絲蛋白mRNA表達(dá)及蛋白含量先下降,但是在產(chǎn)后恢復(fù)期轉(zhuǎn)為上升,微管蛋白mRNA表達(dá)及蛋白含量則先升高,但在產(chǎn)后恢復(fù)期轉(zhuǎn)為下降。 2盆底肌細(xì)胞骨架的變化是對妊娠與分娩引起的肌細(xì)胞牽拉伸展的適應(yīng)性反應(yīng)。 3妊娠與分娩引起的盆底肌細(xì)胞骨架變化在PFD的發(fā)病機(jī)制中有一定的作用。
[Abstract]:Purpose :
1 To investigate the effects of pregnancy and labor on the content of 偽 - actin , actin , synemin and mRNA expression in the levated muscle cells of the rat .
To investigate the relationship between pregnancy and delivery and cytoskeletal .
Objective To investigate the role of cytoskeletal changes in the pathogenesis of pelvic floor dysfunction ( PFD ) in female pelvic floor dysfunction caused by pregnancy and labor .
Method :
1 study : 20 unmated female rats were selected as non - pregnant rats by selecting healthy Wistar rats .
60 pregnant rats were randomly divided into three groups : late pregnancy , postpartum 6 hours , and postpartum 6 days . Each group was 20 . After adaptive feeding , the levator ani muscle was sacrificed at right time . ( supplied by experimental animal part of Hebei Medical University )
2 Materials : The species , litter , age , weight and nutritional status of the rats fed from the experimental animal part of Hebei Medical University were not different from those of food intake . The samples were levator muscle , size about 0.5cm 脳 0.5cm 脳 0.5cm , and the samples were fixed with 10 % neutral formalin and preserved by liquid nitrogen .
3 . The expression of 偽 - actin , micro - tube protein and fibronectin in the levated muscle cells was detected by immunohistochemistry ( IHC SP method ) and real - time fluorescence quantitative polymerase chain reaction ( RT - PCR ) .
4 SPSS 13.0 software was used to analyze the obtained data , which was statistically significant in P0.05 .
Results :
The expression of 偽 - actin ( 偽 - actin ) , microtubular protein ( 偽 - actin ) , microtubular protein ( 偽 - actin ) , fibronectin ( synemin ) , 偽 - actin in non - pregnant group , late pregnancy group , postpartum 6 - hour group and postpartum 6 - day group was significantly higher than that in non - pregnant group ( P < 0.05 ) .
There was significant difference between the three groups ( 1.652 鹵 0.085 , 5.905 鹵 0.13 , 3.059 鹵 0.177 ) and the non - pregnant group ( 1.000 鹵 0.000 ) in the late pregnancy group and the postpartum 6 - hour group ( P < 0.05 ) .
Compared with non - pregnant group ( 0 . 865 鹵 0 . 026,0 . 532 鹵 0.042 , 0.735 鹵 0.014 ) , there was a significant difference between the two groups ( P < 0 . 05 ) .
Conclusion :
The mRNA expression and protein content of 偽 - actin , integrin mRNA and protein in rats induced by pregnancy and labor decreased first , but after the postpartum recovery period , the mRNA expression and protein content of microtubules increased first , but declined during the postpartum recovery period .
The changes of the cytoskeletal structure of the pelvic floor muscle are the adaptive responses to the stretching of the muscle cells caused by pregnancy and labor .
The changes of the cytoskeletal structure of pelvic floor muscle caused by pregnancy and labor play a role in the pathogenesis of PFD .
【學(xué)位授予單位】:河北醫(yī)科大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2014
【分類號(hào)】:R711.5
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