妊娠和分娩对大鼠肛提肌细胞骨架的影响

发布时间：2018-06-21 21:59

本文选题：盆底功能障碍性疾病 + 妊娠　；参考：《河北医科大学》2014年硕士论文

【摘要】：目的： 1研究妊娠和分娩对大鼠肛提肌细胞骨架蛋白α-肌动蛋白（α-actin）、微管蛋白（tubulin）、联丝蛋白（synemin）含量变化及mRNA表达的影响。 2探究妊娠和分娩与细胞骨架的关系。 3探讨妊娠和分娩引起的肛提肌细胞骨架变化在女性盆底功能障碍性疾病（pelvic floor dysfunction，PFD）发病机制中的作用。方法： 1研究对象：选用健康的Wistar育龄大鼠，，取20只未交配的雌鼠作为非妊娠组；再取60只孕鼠随机分为妊娠晚期、产后6小时、产后6天3组，每组20只。适应性喂养后各组均适时处死取肛提肌。(由河北医科大学实验动物部供给) 2取材：由河北医科大学实验动物部供给的大鼠的种属、窝别、年龄、体重和营养状况与进食量等方面无差异。所取标本为肛提肌，大小约0.5cm×0.5cm×0.5cm，所取标本经10%中性福尔马林溶液固定及液氮保存。 3应用免疫组织化学过氧化物酶标记链霉卵白素法（IHC SP法）和实时荧光定量PCR法(RT-PCR)，分别检测非妊娠组、妊娠晚期组、产后6小时组、产后6天组大鼠肛提肌细胞中α-肌动蛋白、微管蛋白、联丝蛋白的表达。 4用SPSS13.0统计软件包，对所得数据进行统计分析，以P0.05为有统计学意义。结果： 1免疫组织化学结果：在非妊娠组、妊娠晚期组、产后6小时组、产后6天组大鼠肛提肌细胞中均检测到α-肌动蛋白（α-actin）、微管蛋白（tubulin）、联丝蛋白（synemin）的表达，α-肌动蛋白在非妊娠组、妊娠晚期组、产后6小时组、产后6天组肛提肌细胞中表达水平的差异具有显著统计学意义（P＜0.05）。微管蛋白在非妊娠组、妊娠晚期组、产后6小时组、产后6天组肛提肌细胞中表达水平的差异具有显著统计学意义（P＜0.05）。联丝蛋白在非妊娠组、妊娠晚期组、产后6小时组、产后6天组肛提肌细胞中表达水平的差异具有显著统计学意义（P＜0.05）。随着妊娠分娩及产后恢复的过程，α-肌动蛋白和联丝蛋白表达量先减少后增加，微管蛋白的表达量先增加后减少，但是组间比较发现，α-肌动蛋白在非妊娠组和妊娠晚期组（P=0.084）中、微管蛋白在妊娠晚期组和产后6天组（P=0.836）中的表达无显著差异性。其余组间比较均有显著差异性。2荧光定量PCR结果：α-肌动蛋白mRNA产后6小时组、产后6天组表达量(0.726±0.021,0.906±0.113)较非妊娠组（1.000±0.000）下降后上升，差异有统计学意义，P＜0.05，妊娠晚期组mRNA表达量（0.975±0.010）下降，与非妊娠组差异无统计学意义，P＞0.05；微管蛋白mRNA妊娠晚期组、产后6小时组、产后6天组表达量(1.652±0.085,5.905±0.13,3.059±0.177)较非妊娠组（1.000±0.000）先升高后下降，各组间差异均有统计学意义，P＜0.05；联丝蛋白mRNA妊娠晚期组、产后6小时组、产后6天组表达量(0.865±0.026,0.532±0.042,0.735±0.014)较非妊娠组（1.000±0.000）先下降后升高，各组间差异均有统计学意义，P＜0.05。结论： 1妊娠与分娩引起大鼠α-肌动蛋白、联丝蛋白mRNA表达及蛋白含量先下降，但是在产后恢复期转为上升，微管蛋白mRNA表达及蛋白含量则先升高，但在产后恢复期转为下降。 2盆底肌细胞骨架的变化是对妊娠与分娩引起的肌细胞牵拉伸展的适应性反应。 3妊娠与分娩引起的盆底肌细胞骨架变化在PFD的发病机制中有一定的作用。
[Abstract]:Purpose :

1 To investigate the effects of pregnancy and labor on the content of 伪 - 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 伪 - 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 伪 - actin ( 伪 - actin ) , microtubular protein ( 伪 - actin ) , microtubular protein ( 伪 - actin ) , fibronectin ( synemin ) , 伪 - 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 卤 0.085 , 5.905 卤 0.13 , 3.059 卤 0.177 ) and the non - pregnant group ( 1.000 卤 0.000 ) in the late pregnancy group and the postpartum 6 - hour group ( P < 0.05 ) .
Compared with non - pregnant group ( 0 . 865 卤 0 . 026,0 . 532 卤 0.042 , 0.735 卤 0.014 ) , there was a significant difference between the two groups ( P < 0 . 05 ) .

Conclusion :

The mRNA expression and protein content of 伪 - 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 .
【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R711.5

【参考文献】