大鼠不稳定膀胱逼尿肌间隙连接蛋白43的含量和分布模式的实验研究

目的探讨对实验大鼠不稳定膀胱逼尿肌连接蛋白43的含量和分布模式进行检测的研究方法。方法实验建立大鼠膀胱出口梗阻模型,并采用充盈性膀胱测压方法确定不稳定膀胱组10只,另设正常对照组10只,应用罗丹明麦芽凝集素标记逼尿肌细胞膜,异硫氰酸荧光(FITC)标记间隙连接蛋白43(Cx43),激光扫描共聚焦显微镜检测Cx43和细胞膜,并用激光共聚焦显微镜系统的Im age Analysis程序对结果进行图像分析。结果在激光扫描共聚焦显微镜下,可以清晰地观察Cx43和细胞膜。不稳定膀胱组Cx43象素密度值明显高于正常对照组(P<0.01);Cx43荧光斑面积也较正常对照组明显增大(P<0.01)。结论双重免疫荧光标记和激光扫描共聚焦显微镜技术能对连接蛋白43进行精确的定量和定位研究,为探讨不稳定膀胱的发生机理提供了一种先进科学的研究方法。

Experimental study of connexin 43 in detrusor of bladder in rats with detrusor instability

Objective To explore the method for detecting the connexin 43(Cx43) in detrusor of bladder in rats with detrusor instability.Methods The model of rats with detrusor instability was established.Ten rats in unstable bladder group were determined by filling cystonetry method controlled by 10 normal rats.The fluorescein isothiocyanate(FITC) was used to label the Cx43 and the tritin conjugated rhodamine was used to label the cell membrane of detrusor muscle.The Cx43 and the cell membrane of detrusor muscle were detected by laser scanning confocal microscopy(LSCM),and the images were analyzed by the image analysis program.Results The distinct images of Cx43 and cell membrane of detrusor muscle could be obtained by the LSCM.The expression of Cx43 and pixel density were increased significantly in rats of unstable bladder group than those of the control group(all P<0.01).Conclusion The quantitation and localization of Cx43 can be precisely detected with the SLCM and double-label immunofluorescence technique,which may serve as a advanced method for determining the mechanism of unstable bladder.