梗阻性不稳定膀胱逼尿肌细胞内Ca2+及N型Ca2+通道的变化及意义

目的对Ca2 和N型Ca2 通道在兔不全梗阻性不稳定膀胱逼尿肌细胞中的变化进行研究。方法成年同龄雄性新西兰白兔30只,随机分两组,15只中膀胱出口不全梗阻8w尿流动力学证实为不稳定膀胱者为实验组,15只为对照组(成年雄性新西兰白兔仅仅手术游离兔膀胱颈而不做结扎梗阻为对照组)。采用急性酶法分离及传代培养的方法获得单个膀胱逼尿肌细胞,采用激光共聚焦显微镜观察模型膀胱逼尿肌细胞静态Ca2 浓度;运用共聚焦显微镜及免疫组织化学方法观察N型Ca2 通道在梗阻性不稳定膀胱逼尿肌细胞中的变化。结果静息状态下不全梗阻性不稳定膀胱组逼尿肌细胞内游离钙离子浓度明显增高(Ca2 超负荷);共聚焦显微镜及免疫组化均证实不稳定膀胱逼尿肌细胞膜之N钙离子通道数量明显增多。结论膀胱逼尿肌细胞Ca2 及其N型Ca2 通道病理性改变是出现不稳定膀胱的重要因素。

Changes in Ca2+ and N-type Ca2+ channel in detrusor muscle cell of obstructively unstable bladder and their significances

Objective To study the changes in Ca2 and N-type Ca2 channel in detrusor muscle cells of the unstable bladder of rabbit's partial bladder outlet obstruction. Methods 30 male New Zealand White rabbits were randomly divided into 2 groups: the experiment group (15 rabbits with partial bladder outlet obstruction lasting for 8 weeks, those with unstable bladder were confirmed by urodynamics) and the control group (15 age-matched New Zealand White rabbits only accepted bladder neck liberation without ligation obstruction as the control group). After established model, bladder detrusor muscle cells of the rabbits were isolated by acute collagenase digestion and serial subcultivation. The static Ca2 concentration in bladder detrusor muscle cells were observed by confocal laser scanning microscope (CLSM). N-type Ca2 channel changes in detrusor muscle cells between the two group was observed by CLSM and immunohistochemistry.Results Ca2 concentration significantly increased in the experimental group (Ca2 overload). The increased N-type Ca2 channels in membrane of bladder detrusor muscle cells were confirmed with CLSM and immunohistochemistry.Conclusions It was an important pathogeny for the unstable bladder of partial bladder outlet obstruction that changes of pathology and pathophysiology of Ca2 and its N-type channels in the unstable bladder.