目的:研究高糖环境下豚鼠膀胱Cajal间质细胞(interstitial cells of Cajal,ICCs)形态及细胞内钙离子荧光的改变,探讨糖尿病膀胱(diabetic cystopathy,DCP)的发病机制。方法:采用酶消化法原代培养,激光共聚焦技术观察5、10、15mol/L葡萄糖浓度下培养24、72h的ICCs,每浓度/时间取30个细胞测量其长度,再从中随机选10个细胞,以200ms/张图片的速度进行扫描,获得每个时间点细胞内钙离子荧光强度值。结果:糖浓度增高培养时间延长,ICCs内钙离子荧光值升高(P=0.00),只有15mol/L/24h降低(P=0.00);ICCs细胞长度缩短(P=0.00),只有10mol/L/72h细胞长度增长(P=0.00)。结论:高糖环境对豚鼠膀胱ICCs形态学及电生理学造成显著影响,这种改变可能是造成DCP发病的重要原因之一。
Objective: To investigate the morphology and electrophysiology changes of the guinea pig bladder Cajal interstitial cells (interstitial cells of Cajal, ICCs) in the high glucose environment, and to investigate the mechanism of diabetic cystopathy(DCP). Method: ICCs bladder cells were cultured by primary culture with collagenase digestion. Confocal laser scanning technology was used to detected the length of ICCs cultured in medium with 5,10,15mol/L glucose and cultured for 24,72h. 10 cells were selected and the calcium fluorescence titer of which were get at the scanning speed of 200ms per slide. Result: The fluorescence titer of Ca2+ in ICCs increased (p=0.00) with the glucose concentration and culture time increased, except cultured 24h in 15 mol/L (p=0.00); The length of ICCs decreased (p=0.000), except cultured 72h in 10mol/1 (p=0.000). Conclusions: High glucose environment significantly affected the morphology and electrophysiology of ICC, and these changes may be one of the most important factor for DCP.