P2Y6受体的激活促进膀胱出口梗阻男性膀胱黏膜下层尿路上皮释放非神经元型三磷酸腺苷和乙酰胆碱。

Activation of P2Y6 Receptors Facilitates Nonneuronal Adenosine Triphosphate and Acetylcholine Release from Urothelium with the Lamina Propria of Men with Bladder Outlet Obstruction.

作者信息

Silva Isabel, Ferreirinha Fátima, Magalhães-Cardoso Maria Teresa, Silva-Ramos Miguel, Correia-de-Sá Paulo

机构信息

Laboratório de Farmacologia e Neurobiologia, Porto, Portugal; Center for Drug Discovery and Innovative Medicines, Porto, Portugal.

Laboratório de Farmacologia e Neurobiologia, Porto, Portugal; Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto and Serviço de Urologia, Centro Hospitalar do Porto, Porto, Portugal.

出版信息

J Urol. 2015 Oct;194(4):1146-54. doi: 10.1016/j.juro.2015.05.080. Epub 2015 May 21.

DOI:10.1016/j.juro.2015.05.080

PMID:26004864

Abstract

PURPOSE

Deregulation of purinergic bladder signaling may contribute to persistent detrusor overactivity in patients with bladder outlet obstruction. Activation of uridine diphosphate sensitive P2Y6 receptors increases voiding frequency in rats indirectly by releasing adenosine triphosphate from the urothelium. To our knowledge this mechanism has never been tested in the human bladder.

MATERIALS AND METHODS

We examined the role of the uridine diphosphate sensitive P2Y6 receptor on tetrodotoxin insensitive nonneuronal adenosine triphosphate and [(3)H]acetylcholine release from the human urothelium with the lamina propria of control organ donors and patients with benign prostatic hyperplasia.

RESULTS

The adenosine triphosphate-to-[(3)H]acetylcholine ratio was fivefold higher in mucosal urothelium/lamina propria strips from benign prostatic hyperplasia patients than control men. The selective P2Y6 receptor agonist PSB0474 (100 nM) augmented by a similar amount adenosine triphosphate and [(3)H]acetylcholine release from mucosal urothelium/lamina propria strips from both groups of individuals. The facilitatory effect of PSB0474 was prevented by MRS2578 (50 nM) and by carbenoxolone (10 μM), which block P2Y6 receptor and pannexin-1 hemichannels, respectively. Blockade of P2X3 (and/or P2X2/3) receptors with A317491 (100 nM) also attenuated release facilitation by PSB0474 in control men but not in patients with benign prostatic hyperplasia. Immunolocalization studies showed that P2Y6, P2X2 and P2X3 receptors were present in choline acetyltransferase positive urothelial cells. In contrast to P2Y6 staining, choline acetyltransferase, P2X2 and P2X3 immunoreactivity decreased in the urothelium of benign prostatic hyperplasia patients.

CONCLUSIONS

Activation of P2Y6 receptor amplifies mucosal adenosine triphosphate release underlying bladder overactivity in patients with benign prostatic hyperplasia. Therefore, we propose selective P2Y6 receptor blockade as a novel therapeutic strategy to control persistent storage symptoms in obstructed patients.

摘要

目的

嘌呤能膀胱信号传导失调可能导致膀胱出口梗阻患者持续性逼尿肌过度活动。尿苷二磷酸敏感的P2Y6受体激活通过从尿路上皮释放三磷酸腺苷间接增加大鼠排尿频率。据我们所知，这种机制从未在人膀胱中得到验证。

材料与方法

我们研究了尿苷二磷酸敏感的P2Y6受体对来自对照器官供体和良性前列腺增生患者的人尿路上皮与固有层中河豚毒素不敏感的非神经元性三磷酸腺苷和[³H]乙酰胆碱释放的作用。

结果

良性前列腺增生患者黏膜尿路上皮/固有层条带中三磷酸腺苷与[³H]乙酰胆碱的比率比对照男性高五倍。选择性P2Y6受体激动剂PSB0474（100 nM）使两组个体黏膜尿路上皮/固有层条带中的三磷酸腺苷和[³H]乙酰胆碱释放量增加相似的幅度。MRS2578（50 nM）和羧苄青霉素（10 μM）分别阻断P2Y6受体和泛连接蛋白-1半通道，从而阻止了PSB0474的促进作用。用A317491（100 nM）阻断P2X3（和/或P2X2/3）受体也减弱了PSB0474对对照男性释放的促进作用，但对良性前列腺增生患者没有作用。免疫定位研究表明，P2Y6、P2X2和P2X3受体存在于胆碱乙酰转移酶阳性的尿路上皮细胞中。与P2Y6染色不同，良性前列腺增生患者尿路上皮中的胆碱乙酰转移酶、P2X2和P2X3免疫反应性降低。