CX43蛋白磷酸化在大鼠逼尿肌过度活动症中的作用.docVIP

DOI：10.16016/j.1000-5404.201411215 CX43蛋白磷酸化在大鼠逼尿肌过度活动症中的作用 朱景振1，刘 骞2 ，董兴有2，龙 州2，李龙坤2，宋 波1 （400038重庆，第三军医大学西南医院全军泌尿外科研究所1；400037 重庆，第三军医大学新桥医院泌尿外科2） [摘要] 目的 评价CX43蛋白磷酸化在大鼠逼尿肌过度活动症（detrusor overactivity, DO）中的作用，并初步探讨PP1和PP2A对CX43蛋白磷酸化修饰状态的影响。方法 建立大鼠DO模型作为实验组，假手术组作为对照组，Western blot检测CX43蛋白磷酸化水平、PP1及PP2A的表达变化，免疫荧光双染对CX43与PP1、PP2A的定位及表达变化进行检测，荧光漂白恢复实验（FRAP）比较正常组、DO组、DO+碱性磷酸酶（AP）组的细胞间通讯功能（GJIC）的差异。结果 与正常组相比，DO组膀胱中的CX43蛋白的不同磷酸化修饰状态（P0、P1、P2）的表达水平均明显升高（P＜0.05），PP2A的表达明显降低（P＜0.05），PP1的表达未发现明显改变（P＞0.05）。DO组的GJIC较正常组明显增加（P＜0.05），DO+AP组的GJIC较正常组和DO组明显降低（P＜0.05）。结论 CX43蛋白的磷酸化水平上调可能在DO的发病过程中发挥着重要的作用，这种磷酸化水平的上调可能与PP2A的表达减少相关。 [关键词] CX43蛋白；磷酸化；逼尿肌过度活动症 [中图法分类号] R322.62；R694.52；R334.3 [文献标志码] A Effect of Connexin 43 phosphorylation on detrusor overactivity in rats Zhu Jingzhen1, Liu Qian2, Dong Xingyou2, Long Zhou2, Li Longkun2, Song Bo1 (Research Institute of Urinary Surgery1, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China; Department of Urology2, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China) [Abstract] Objective To evaluate the effect of Connexin 43 (CX43) phosphorylation on detrusor overactivity (DO) in rats, and preliminarily investigate the influence of protein phosphatase 1 (PP1) and protein phosphatase 2A (PP2A) on CX43 phosphorylation. Methods DO rat model was established as the experimental group, and sham-operated rats were selected as the control group. The CX43 protein phosphorylation levels and the PP1 and PP2A expression changes were detected by Western blot. Double immunofluorescence staining was used for detecting the localization and expression changes of CX43, PP1 and PP2A. Fluorescence recovery after photobleaching (FRAP) was employed to compare the gap junction intercellular communication (GJIC) of the control group, the DO group and the DO+alkaline phosphatase (AP) group. Results Compared with the control group, the expression of CX43 protein with different phosphor