本文關(guān)鍵詞：大鼠妊娠與分娩期盆底肌中Desmin及Dystrophin的表達及意義　出處：《河北醫(yī)科大學》2014年碩士論文　論文類型：學位論文

  更多相關(guān)文章： 盆底功能障礙性疾病 細胞骨架 結(jié)蛋白（Desmin） 抗肌營養(yǎng)不良蛋白（Dystrophin） 妊娠與分娩

【摘要】：目的: 1研究大鼠妊娠與分娩對肛提肌中結(jié)蛋白（Desmin）與抗肌營養(yǎng)不良蛋白（Dystrophin）的分布及其基因表達量的影響。 2研究大鼠妊娠與分娩對盆底肌細胞骨架的影響。 3探討妊娠與分娩在盆底功能障礙性疾�。╬elvic floor dysfunction, PFD）發(fā)病機制中的作用。 方法: 1研究對象由河北醫(yī)科大學實驗動物中心提供雌性成年未生產(chǎn)過的Wistar大鼠45只，體重180～220g。雄性成年Wistar大鼠10只，體重220～250g。其中隨機選取15只雌性成年未生產(chǎn)過的Wistar大鼠為對照組，其余30只雌性成年未生產(chǎn)過的Wistar大鼠使其受孕并分娩，分為陰道分娩組，產(chǎn)后6天組。 2取材將各組大鼠在麻醉狀態(tài)下，，暴露肛提肌，迅速取肛提肌組織約100mg。分為兩塊，生理鹽水洗凈，用濾紙吸干，一塊置于凍存管，立即置-80℃低溫凍存，用于實時熒光定量PCR測定；另1塊置入體積分數(shù)為4%多聚甲醛溶液中固定24h，常規(guī)脫水、石蠟包埋備用。 3采用HE染色及免疫組織化學方法觀察各組肛提肌形態(tài)的變化及Desmin和Dystrophin分布變化。 4采用實時熒光定量PCR檢測各組肛提肌中Desmin和Dystrophin基因表達量的變化。結(jié)果資料采用均數(shù)±標準差（±S）表示，采用SPSS16.0統(tǒng)計軟件進行統(tǒng)計學處理，以P＜0.05認為差異有統(tǒng)計學意義。 結(jié)果： 1HE染色結(jié)果：HE染色鏡下觀察各組肛提肌切片，其中對照組肛提肌組織均由彼此緊密相連的肌纖維構(gòu)成，為均染的粉紅色，每個肌纖維周邊有多個細胞核，緊靠肌膜內(nèi)表面，肌束間含有膠原纖維、成纖維細胞，血管及神經(jīng)等結(jié)構(gòu)，肌纖維無變性壞死表現(xiàn)；而陰道分娩組肛提肌肌纖維較對照組肥大，排列相對稀疏，略微紊亂，可見少部分肌細胞橫截面積大小不一、有萎縮和肥大細胞，局部可見到肌纖維核中心移位、存在散在壞死肌纖維等；產(chǎn)后6天組肛提肌肌纖維排列較緊密，肌纖維無變性壞死表現(xiàn)。 2免疫組化染色結(jié)果：免疫組化Desmin陽性反應(yīng)位于肌纖維胞漿內(nèi)，而Dystrophin陽性反應(yīng)則位于肌纖維胞膜。陰道分娩組Desmin的表達與非妊娠組和產(chǎn)后6天組相比明顯減少，差異具有統(tǒng)計學意義（P＜0.05），而Desmin的表達在非妊娠組和產(chǎn)后6天組之間無顯著差異；陰道分娩組Dystrophin的表達較非妊娠組和產(chǎn)后6天組減少，差異具有統(tǒng)計學意義（P＜0.05），非妊娠組和產(chǎn)后6天組之間無顯著差異。3熒光實時定量PCR結(jié)果：陰道分娩組Desmin mRNA表達量較非妊娠組、產(chǎn)后6天組明顯降低，差異均有統(tǒng)計學意義，P＜0.05，產(chǎn)后6天組Desmin mRNA表達量與非妊娠組差異無統(tǒng)計學意義；陰道分娩組Dystrophin mRNA表達量較非妊娠組、產(chǎn)后6天組下降，差異均有統(tǒng)計學意義，P＜0.05，而非妊娠組與產(chǎn)后6天組差異無統(tǒng)計學意義。 結(jié)論： 1在大鼠妊娠分娩期盆底肌纖維形態(tài)發(fā)生改變，并引起Desmin、Dystrophin表達減少，在產(chǎn)后盆底肌纖維發(fā)生再生，Desmin、Dystrophin合成增加。 2盆底肌細胞骨架改變是對妊娠與分娩的適應(yīng)性反應(yīng)。 3妊娠與分娩在盆底功能障礙性疾病（pelvic floor dysfunction, PFD）的發(fā)病機制中有一定的作用。妊娠與分娩期大鼠盆底肌中Desmin和dystrophin減少可能是盆底功能障礙性疾病的發(fā)病因素。
[Abstract]:Objective:
1 the effects of pregnancy and childbirth on the distribution of Desmin and Dystrophin in the muscle of the anal levator and the expression of gene expression in the muscle of the anal levator were studied.
2 the effects of pregnancy and childbirth on the cytoskeleton of pelvic floor muscle in rats were studied.
3 to investigate the role of pregnancy and childbirth in the pathogenesis of pelvic floor dysfunction (PFD).
Method:
1 research objects provided by the experimental animal center of Hebei Medical University female adult nulliparous 45 Wistar rats, weighing 180 ~ 220g. adult male Wistar 10 rats, Wistar rats weighing 220 ~ 250g. were randomly selected 15 adult nulliparous females as control group, Wistar rats had not produced the remaining 30 the adult female pregnancy and childbirth, divided into the vaginal delivery group, postpartum 6 days group.
2 were the rats under anesthesia, exposed levator ani muscle, quickly take levator muscle tissue about 100mg. is divided into two pieces, saline wash, dry with filter paper, a block on the freezing tube, at -80 deg.c for cryopreservation, real-time fluorescence quantitative PCR assay; the other 1 implants the integral number of 4% poly fixed 24h, Formaldehyde Solution in the conventional dehydration, paraffin embedded sections.
3 HE staining and immunohistochemical method were used to observe the changes of the morphology of the levator anus and the changes of the distribution of Desmin and Dystrophin.
4, real-time fluorescence quantitative PCR was used to detect the changes of Desmin and Dystrophin gene expression in each levator ani muscle. Results the data were expressed by mean + standard deviation (+ S), and SPSS16.0 statistics software was used for statistical analysis. The difference was statistically significant in P < 0.05.
Result錛

本文編號：1440374