大鼠不稳定膀胱中ICCs细胞中HCN蛋白表达的变化  

Variation of HCN protein expression in interstitial cells of Cajal from rat with detrusor instability

作　　者：丁砺蠡[1] 王永权[1] 方强[1] 封建立[1] 李龙坤[1] 宋波[1] 

机构地区：[1]第三军医大学西南医院全军泌尿外科研究所,重庆400038

出　　处：《第三军医大学学报》2008年第7期578-581,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30700270;30500505)~~

摘　　要：目的研究HCN离子通道(hyperpolarization-activated cyclic nucleotide-gated cation channel,HCN)蛋白在膀胱Cajal间质细胞(interstitial cells of Cajal,ICCs)中的表达及在不稳定膀胱ICCs细胞中表达的变化,探讨ICCs细胞兴奋性变化与逼尿肌不稳定(detrusor instability,DI)的关系。方法①将30只大鼠随机分为正常对照(10例),逼尿肌稳定组(DS,n=12),逼尿肌不稳定组(DI,n=8):大鼠尿道近端结扎,建立膀胱出口梗阻(bladder outlet obstruction,BOO)模型。②对各组进行HCN和c-kit免疫荧光双标,确定HCN蛋白是否在ICCs细胞上存在及HCN表达的变化。③以Western blot法检测HCN蛋白表达量的变化。结果①免疫荧光双标显示,在c-kit阳性的ICCs细胞膜上,有HCN抗原的表达,c-kit阴性细胞未见HCN表达;与正常组相比,DI组、DS组HCN蛋白表达增加。②Western blot检测显示,与正常组相比,DI组、DS组的HCN表达增高,DI组增高显著(P<0.01)。结论大鼠膀胱ICCs细胞可表达HCN通道蛋白,且DI膀胱ICCs细胞HCN通道明显增多,由此导致的ICCs细胞兴奋性增高可能在DI发生中起重要作用。Objective To investigate whether there is expression of hyperpolarization-activated cyclic nucleotide-gated cation channel （HCN） in normal bladder interstitial cells of Cajal （ICCs） and in ICCs of unstable bladder, so as to assess the relationship between the changes of ICCs excitability and the detrusor instability. Methods （1） Thirty SD rats were randomized into 3 groups： normal （n = 10）, detrusor stability （n = 12） and detrusor instability （ n = 8, 8 weeks after bladder outlet obstruction）. （2） The HCN protein expression in ICCs was detected with double-labeled immunofluorescence. The relationship of HCN protein and ICCs was accessed by double-labeled immunofluorescence with HCN and c-kit. （3） HCN expression of the 3 groups was also detected with Western blotting assay. Results （1） HCN expression was well accorded to c-kit expression by using double-labeled immunofluorescence. （2） With double-labeled immunofluorescence and Western blotting, HCN expression of DI and stable group was increased compared to the normal group, with statistical significance between the DI and normal groups （P 〈 0.01 ）. Conclusion HCN protein is identified in ICCs of rat bladder. The expression of HCN protein is enhanced obviously in DI group, which may be responsible for the increased excitability of DI bladder.

关 键 词：ICCS细胞 HCN通道 逼尿肌不稳定 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]