Bladder Urothelial Cells From Patients With Interstitial Cystitis Have an Increased Sensitivity to Carbachol
Bladder Urothelial Cells From Patients With Interstitial Cystitis Have an Increased Sensitivity to Carbachol
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DOI：
10.1002/nau.20726
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发表时间：
2009-01-01
影响因子：
2
通讯作者：
Chail, Toby C.
中科院分区：
文献类型：
--
作者：
Gupta, Gopal N.;Lu, Shao-Gang;Chail, Toby C.
目的：膀胱尿路上皮细胞（BUC）上存在毒蕈碱受体，提示BUC可能是抗毒蕈碱的靶点。本研究确定了人BUC是否对毒蕈碱激动剂有反应，如果有反应，是否在间质性膀胱炎（IC） BUC中发生改变。方法：采用膀胱镜活检法培养原代尿路上皮细胞。采用钙显像技术研究了正常（NB）和IC BUC，以监测激动剂恰巴醇（CCh）对毒蕈碱受体激活的反应。细胞内Ca2+浓度的变化[（Ca2+](i)）测量fura-2比例微荧光法。在IC和NB BUC中测量剂量-反应曲线（CCh剂量vs [Ca2+](i)）。托特罗定被用来确认CCh引起的[Ca2+]增加的特异性（毒蕈碱与烟碱）(i)。结果：CCh诱导[Ca2+]的剂量依赖性增加(i)。CCh在IC BUC中的效价和疗效显著提高。在4例IC患者的78个细胞中，[Ca2+](i)比基线增加了136.3 +/- 5.1%，而在4例NB患者的67个细胞中，[Ca2+](i)比基线增加了92.4 +/- 4.8% （P < 0.01）。对照和NB组BUC的EC50分别为1.10 +/- 0.14 μ M和3.36 +/- 0.72 μ M （P < 0.01）。去除细胞外钙或应用托特罗定，消除CCh引起IC和NB BUC中[Ca2+](i)的增加。结论：IC BUC对CCh的更大敏感性表明IC的病理生物学也可能包括毒蕈碱信号的改变。毒蕈碱在BUC中活化的生理后遗症有待进一步研究。Neurourol。生物工程学报，28(2):1022-1027,2009。(C) 2009 Wiley-Liss, Inc。
Aims: The presence of muscarinic receptors on bladder urothelial cells (BUC), suggests BUC may be a target for antimuscarinics. This study determined whether human BUC are responsive to a muscarinic agonist and if so, whether responses are altered in interstitial cystitis (IC) BUC. Methods: Primary urothelial cell cultures were established from cystoscopic biopsies. Normal (NB) and IC BUC were studied using calcium imaging techniques as a means to monitor the response to muscarinic receptor activation with the agonist, carbachol (CCh). Changes in intracellular Ca2+ concentration [(Ca2+](i)) were measured with fura-2 ratiometric microfluorimetry. Dose-response curves (CCh dose vs. [Ca2+](i)) were measured in IC and NB BUC. Tolterodine was used to confirm the specificity (muscarinic versus nicotinic) of CCh evoked increases in [Ca2+](i). Results: CCh induced a dose-dependent increase in [Ca2+](i). Potency and efficacy of CCh was significantly greater in IC BUC. The maximal increase in [Ca2+](i) was 136.3 +/- 5.1% over baseline in 78 cells from 4 IC patients versus 92.4 +/- 4.8% over baseline in 67 cells from 4 NB subjects (P < 0.01). The EC50 of the evoked increase was 1.10 +/- 0.14 mu M versus 3.36 +/- 0.72 mu M (P < 0.01) in BUC from IC and NB controls, respectively. Removal of extracellular calcium or application of tolterodine, abolished CCh evoked increase in [Ca2+](i) in IC and NB BUC. Conclusions: The greater sensitivity of IC BUC to CCh suggests that IC patholobiology may also include alterations muscarinic signaling. The physiologic sequelae of muscarinic activation in BUC need to be further investigated. Neurourol. Urodynam. 28:1022-1027, 2009. (C) 2009 Wiley-Liss, Inc.