The James Buchanan Brady Urological Institute and Department of Urology, The Johns Hopkins, University School of Medicine, Baltimore, Maryland (S.K., B.M., A.L.B.); and Departments of Physiology and Biophysics (M.S.N., J.M.F., K.P.D.) and Urology (K.P.D.), Albert Einstein College of Medicine, New York, New York
The James Buchanan Brady Urological Institute and Department of Urology, The Johns Hopkins, University School of Medicine, Baltimore, Maryland (S.K., B.M., A.L.B.); and Departments of Physiology and Biophysics (M.S.N., J.M.F., K.P.D.) and Urology (K.P.D.), Albert Einstein College of Medicine, New York, New York.
J Pharmacol Exp Ther. 2020 May;373(2):214-219. doi: 10.1124/jpet.119.264697. Epub 2020 Mar 6.
Sickle cell disease (SCD) is associated with overactive bladder (OAB). Detrusor overactivity, a component of OAB, is present in an SCD mouse, but the molecular mechanisms for this condition are not well-defined. We hypothesize that nitric oxide (NO)/ ras homolog gene family (Rho) A/Rho-associated kinase (ROCK) dysregulation is a mechanism for detrusor overactivity and that NO-releasing nanoparticles (NO-nps), a novel NO delivery system, may serve to treat this condition. Male adult SCD transgenic, combined endothelial NO synthases (eNOSs) and neuronal NOS (nNOS) gene-deficient (dNOS), and wild-type (WT) mice were used. Empty nanoparticle or NO-np was injected into the bladder, followed by cystometric studies. The expression levels of phosphorylated eNOS (Ser-1177), protein kinase B (Akt) (Ser-473), nNOS (Ser-1412), and myosin phosphatase target subunit 1 (MYPT1) (Thr-696) were assessed in the bladder. SCD and dNOS mice had a greater ( < 0.05) number of voiding and nonvoiding contractions compared with WT mice, and they were normalized by NO-np treatment. eNOS (Ser-1177) and AKT (Ser-473) phosphorylation were decreased ( < 0.05) in the bladder of SCD compared with WT mice and reversed by NO-np. Phosphorylated MYPT1, a marker of the RhoA/ROCK pathway, was increased ( < 0.05) in the bladder of SCD mice compared with WT and reversed by NO-np. nNOS phosphorylation on positive (Ser-1412) regulatory site was decreased ( < 0.05) in the bladder of SCD mice compared with WT and was not affected by NO-np. NO-nps did not affect any of the measured parameters in WT mice. In conclusion, dysregulation of NO and RhoA/ROCK pathways is associated with detrusor overactivity in SCD mice; NO-np reverses these molecular derangements in the bladder and decreases detrusor overactivity. SIGNIFICANCE STATEMENT: Voiding abnormalities commonly affect patients with sickle cell disease (SCD) but are problematic to treat. Clarification of the science for this condition in an animal model of SCD may lead to improved interventions for it. Our findings suggest that novel topical delivery of a vasorelaxant agent nitric oxide into the bladder of these mice corrects overactive bladder by improving deranged bladder physiology regulatory signaling.
镰状细胞病 (SCD) 与膀胱过度活动症 (OAB) 有关。逼尿肌过度活动是 OAB 的一个组成部分，在 SCD 小鼠中存在，但这种情况的分子机制尚不清楚。我们假设一氧化氮 (NO)/ras 同源基因家族 (Rho) A/Rho 相关激酶 (ROCK) 失调是逼尿肌过度活动的一种机制，而释放一氧化氮的纳米粒子 (NO-nps) 作为一种新型的一氧化氮输送系统，可能有助于治疗这种疾病。使用雄性成年 SCD 转基因、联合内皮型一氧化氮合酶 (eNOS) 和神经元型一氧化氮合酶 (nNOS) 基因缺陷 (dNOS) 和野生型 (WT) 小鼠。将空纳米颗粒或 NO-np 注入膀胱，然后进行膀胱测压研究。评估膀胱中磷酸化 eNOS (Ser-1177)、蛋白激酶 B (Akt) (Ser-473)、nNOS (Ser-1412) 和肌球蛋白磷酸酶靶亚单位 1 (MYPT1) (Thr-696) 的表达水平。与 WT 小鼠相比，SCD 和 dNOS 小鼠的排尿和非排尿收缩次数更多（<0.05），并且通过 NO-np 治疗得到了正常化。与 WT 小鼠相比，SCD 小鼠的 eNOS (Ser-1177) 和 AKT (Ser-473) 磷酸化减少（<0.05），并通过 NO-np 逆转。磷酸化的 MYPT1，RhoA/ROCK 途径的标志物，在 SCD 小鼠的膀胱中增加（<0.05），并通过 NO-np 逆转。与 WT 小鼠相比，SCD 小鼠膀胱中 nNOS 磷酸化（正调节位点 Ser-1412）减少（<0.05），并且不受 NO-np 影响。NO-np 对 WT 小鼠的任何测量参数均无影响。总之，NO 和 RhoA/ROCK 途径的失调与 SCD 小鼠的逼尿肌过度活动有关；NO-np 逆转了膀胱中的这些分子异常，并减少了逼尿肌过度活动。意义声明：排尿异常常影响镰状细胞病 (SCD) 患者，但难以治疗。在 SCD 的动物模型中阐明这种疾病的科学可能会为其带来更好的干预措施。我们的研究结果表明，将血管舒张剂一氧化氮通过新型局部递送至这些小鼠的膀胱中，可以通过改善膀胱生理调节信号来纠正过度活跃的膀胱。
