Akino Hironobu, Chapple Christopher R, McKay Neil, Cross Rebecca L, Murakami Shigetaka, Yokoyama Osamu, Chess-Williams Russell, Sellers Donna J
Department of Biomedical Science, University of Sheffield, Sheffield, UK.
BJU Int. 2008 Nov;102(9):1168-74. doi: 10.1111/j.1464-410X.2008.07782.x. Epub 2008 Jul 21.
To investigate the influence of the mucosa on the inhibitory effects of the ATP-sensitive potassium channel (K(ATP) channel) opener, cromakalim, on the spontaneous contractions of pig bladder strips from the bladder dome and trigone. Little is known about the influence of the mucosa on spontaneous contractions and whether the nature of these contractions differs between the bladder dome and trigone.
Paired longitudinal strips of female pig bladders were isolated from the dome and trigone. The mucosa was removed from one strip per pair and tissues were set up in organ baths. Spontaneous activity was allowed to develop and recorded, and then cumulative concentration-response curves to cromakalim were obtained. The time needed for spontaneous contractions to develop, the frequency and amplitude of spontaneous contractions, and the effect of cromakalim were analysed. The strips of mucosa removed from the dome to produce denuded strips were also analysed by immunofluorescence using antibodies specific for vimentin and alpha-smooth muscle actin (alpha-SMA).
In the dome removal of the mucosa delayed the development of spontaneous contractions compared with mucosa-intact strips, whilst the trigone strips developed spontaneous contractions soon after set up in the organ baths irrespective of the presence or absence of mucosa. In the dome, cromakalim was more potent in suppressing spontaneous contractions when the mucosa was absent; whilst in the trigone the effects of cromakalim were similar in mucosa-intact and denuded strips. Upon examination of the strips of mucosa by immunofluorescence these strips were shown to contain cells positive for alpha-SMA or vimentin and cells positive for both, suggesting the presence of not only urothelium but also suburothelium and some detrusor smooth muscle bundles.
In the dome, the urothelium and suburothelium reduce the inhibitory effect of cromakalim on spontaneous contractions, whilst in the trigone these structures appear to have little influence. The mechanism for generating spontaneous contractions in the intact strips seems to be linked to the urothelium and suburothelium in the dome but not in the trigone.
研究黏膜对三磷酸腺苷敏感性钾通道(K(ATP)通道)开放剂克罗卡林抑制猪膀胱顶部和三角区条带自发收缩作用的影响。关于黏膜对自发收缩的影响以及膀胱顶部和三角区这些收缩的性质是否不同,目前所知甚少。
从雌性猪膀胱分离出成对的纵向条带,分别取自膀胱顶部和三角区。每对条带中一条去除黏膜,将组织置于器官浴槽中。记录自发活动,然后获得克罗卡林的累积浓度-反应曲线。分析自发收缩产生所需时间、自发收缩的频率和幅度以及克罗卡林的作用。对从膀胱顶部去除黏膜后得到的去上皮条带,也使用波形蛋白和α-平滑肌肌动蛋白(α-SMA)特异性抗体通过免疫荧光进行分析。
在膀胱顶部,与保留黏膜的条带相比,去除黏膜会延迟自发收缩的产生,而三角区条带在置于器官浴槽后很快就会产生自发收缩,无论有无黏膜。在膀胱顶部,去除黏膜时克罗卡林抑制自发收缩的作用更强;而在三角区,克罗卡林在保留黏膜和去上皮条带中的作用相似。通过免疫荧光检查黏膜条带发现,这些条带含有α-SMA或波形蛋白阳性细胞以及两者均阳性的细胞,这表明不仅存在尿路上皮,还存在尿路上皮下层和一些逼尿肌平滑肌束。
在膀胱顶部,尿路上皮和尿路上皮下层会降低克罗卡林对自发收缩的抑制作用,而在三角区这些结构似乎影响不大。完整条带中产生自发收缩的机制似乎与膀胱顶部的尿路上皮和尿路上皮下层有关,而与三角区无关。