Li Mingkai, Sun Yan, Simard J Marc, Wang Jian-Ying, Chai Toby C
Department of Surgery, University of Maryland School of Medicine, Baltimore, 21201, USA.
Am J Physiol Cell Physiol. 2009 Dec;297(6):C1445-51. doi: 10.1152/ajpcell.00259.2009. Epub 2009 Oct 7.
Overactive bladder syndrome (OAB) is an idiopathic condition characterized by urinary urgency and urge incontinence. Detrusor overactivity has been traditionally described as the physiologic mechanism for OAB. However, the bladder urothelium (BU) may also be involved in the pathophysiology. This study measured polyamine signaling and its downstream effects on membrane conductivity in bladder urothelial cells (BUC) obtained from asymptomatic and OAB subjects. Immunohistofluorescence was used to measure ornithine decarboxylase (ODC) expression in BU. BUC, cultured from BU biopsies, were used for electrophysiologic studies. dl-alpha-Difluoromethylornithine (DFMO), spermine, or spermidine was used to modulate polyamine signaling in BUC. Results showed ODC overexpression in OAB BU. In OAB BUC, whole cell and cell-attached configuration showed significantly decreased currents. Using inside-out patches, outward currents increased significantly, suggesting a cytoplasmic source of the outward current block in OAB BUC. In control BUC, outward currents were mediated by the large-conductance calcium-activated potassium (BK) channel due to calcium dose-dependence and block by iberiotoxin. Spermidine and spermine blocked the outward current in normal BUC in dose-dependent fashion. Conversely, DFMO significantly increased (P < 0.01) outward currents in OAB BUC both in cell-attached and in whole cell configuration. The outward currents in DFMO-treated-OAB BUC could be significantly reduced (P < 0.05) by adding back spermidine and spermine. These data suggest that polyamine signaling is upregulated in OAB urothelium and OAB BUC. Furthermore, polyamines in BUC block the BK channel. Targeting of bladder urothelial polyamine signaling may represent a novel approach for OAB treatment based on pathophysiologic mechanisms.
膀胱过度活动症(OAB)是一种以尿急和急迫性尿失禁为特征的特发性疾病。传统上认为逼尿肌过度活动是OAB的生理机制。然而,膀胱尿路上皮(BU)也可能参与其病理生理过程。本研究测量了从无症状和OAB受试者获取的膀胱尿路上皮细胞(BUC)中的多胺信号及其对膜电导率的下游影响。采用免疫组织荧光法测量BU中鸟氨酸脱羧酶(ODC)的表达。从BU活检组织培养的BUC用于电生理研究。使用dl-α-二氟甲基鸟氨酸(DFMO)、精胺或亚精胺调节BUC中的多胺信号。结果显示OAB的BU中ODC过表达。在OAB的BUC中,全细胞和细胞贴附模式显示电流显著降低。使用内面向外的膜片,外向电流显著增加,表明OAB的BUC中外向电流阻滞的细胞质来源。在对照BUC中,外向电流由大电导钙激活钾(BK)通道介导,这是由于钙剂量依赖性和iberiotoxin的阻断作用。亚精胺和精胺以剂量依赖性方式阻断正常BUC中的外向电流。相反,DFMO在细胞贴附和全细胞模式下均显著增加(P < 0.01)OAB的BUC中的外向电流。通过添加亚精胺和精胺,DFMO处理的OAB的BUC中的外向电流可显著降低(P < 0.05)。这些数据表明,OAB尿路上皮和OAB的BUC中的多胺信号上调。此外,BUC中的多胺阻断BK通道。基于病理生理机制,靶向膀胱尿路上皮多胺信号可能代表一种治疗OAB的新方法。