用于尿功能测试的成人男性尿道超声检查
Summary
我们描述了使用高频超声与对比成像作为测量膀胱体积、膀胱壁厚度、尿速、空隙体积、空隙持续时间和尿道直径的方法。此策略可用于评估下尿路功能障碍 (LUTD) 的各种小鼠模型中的无效功能障碍和治疗效果。
Abstract
由于人口老龄化,临床良性前列腺增生(BPH)和下尿路症状(LUTS)的发病率呈上升趋势,造成了严重的经济和质量负担。转基因和其他小鼠模型已经开发,以重现这种多因素疾病的各个方面;然而,缺乏准确量化尿功能障碍的方法和新的治疗方案的有效性。在这里,我们描述了一种可用于测量膀胱体积和分泌物壁厚、尿速、空隙体积和空隙持续时间以及尿道直径的方法。这将允许评估疾病进展和治疗效果随着时间的推移。小鼠用胶质麻醉,用超声波对膀胱进行可视化。对于非对比度成像,对膀胱进行三维图像,以计算体积和评估形状;从这张图像测量了膀胱壁厚度。为了进行对比度增强成像,使用 PE50 管连接到注射器的 27 号针在膀胱圆顶上放置导管。在排尿事件发生之前,将0.5 mL的对比波勒注入膀胱。在第一次空隙事件中,在多普勒速度样品窗口的点处确定了尿道直径。对产生流速的每个后续事件测量速度。最后,高频超声证明是评估小鼠泌尿功能期间膀胱和尿道测量的有效方法。此技术在实验环境中评估 BPH/LUTS 的新疗法可能很有用。
Introduction
良性前列腺增生(BPH)是一种随着男性年龄增长而发展的疾病,影响近90%的80岁以上男性1、2岁。虽然BPH的发展一般与衰老有关,其他因素,包括肥胖和代谢综合征,可导致BPH在相对年轻的男性3,4。许多患有BPH的男性出现下尿道症状(LUTS),这大大降低了他们的生活质量,有些症状并发症可能包括出血、感染、膀胱出口阻塞(BOO)、膀胱结石和肾衰竭。BPH的治疗费用每年超过40亿美元。BPH引起的LUTS诊断通常依赖于使用AUA症状指数(AUASI)评分,尿流测量,以及前列腺大小8的评估。BPH/LUTS 的病因是复杂和多因素的,疾病的发展和进展与前列腺增生(前列腺增殖)、平滑肌肉收缩和纤维化相关。目前的治疗包括使用β-肾上腺素阻滞剂调节膀胱和前列腺内的平滑肌肉张力,以减轻LUTS和/或5+还原酶抑制剂,以减少雄激素代谢和减少前列腺大小。更好的疾病模型,鼠等,使准确的研究各种因果关系和治疗因素的影响,在这种疾病过程中是极可取的9。
啮齿动物模型已被广泛用于研究泌尿动力学;然而,大多数研究都集中在女性云母和疾病10。为了全面检查雄性LUTS的各个方面,已经开发并用于研究BPH的不同方面,包括细胞增殖的变化,平滑的肌肉功能,胶原蛋白沉积和炎症11。12,13,然而,啮齿动物和人类前列腺解剖学不同.虽然人类前列腺是紧凑的,包裹在冷凝纤维肌肉层,啮齿动物前列腺是球状;这些差异使疾病进展和治疗效果的直接比较复杂化。此外,LUTS在小鼠中很难评估,因为无法直接测量麻烦。相反,目前研究疾病的方法将组织特征与比较泌尿水平的生理特征(即膀胱体积和壁厚与尿流测定、空隙点测定和膀胱学终点数据)相关联BPH模型和控制动物之间的功能障碍12,15,16,17,18。生理特征经常被评估为死后的尸检终点,并且在同一动物内无法跨时间观察BOO。最近,我们确定了骨盆尿道(前列腺尿道)的细分,其中外源性激素植入物导致缩小基于验尸后尸检评估12。目前的方法不允许在空隙期间对尿道缩小进行直接的体内评估。
超声波是一种非侵入性诊断和评估技术,已成功应用于其他疾病模型。它用于量化器官体积和评估血管流量19,20,21。超声波还用于可视化和指导显微注射,允许有针对性地注射干细胞或其他药物,并评估收缩和舒张心脏功能。
该协议描述了使用高频超声波来评估下尿道解剖学和评估麻醉小鼠的泌尿生理学。我们描述了超声波用于测量膀胱体积和壁厚。我们还描述了使用对比度增强的超声波来测量尿速、尿量、空隙持续时间和尿道直径。超声波的使用提供了对体内下尿道的更全面的了解,确定疾病如何改变正常的空隙功能,并为我们提供了更好的评估新治疗方案有效性的工具。目前,非对比度成像协议是非终端,而当前对比度增强成像协议是一个终端程序。
Protocol
Representative Results
Discussion
目前评估啮齿动物下尿路的技术受限于它们将空隙生理学的变化与导致疾病进展的前列腺动物学变化直接关联的能力。虚斑测定和尿流测定可用于评估啮齿动物的自发排尿事件,这些技术可用于评估15、16、17期间的变化。然而,对于这两种技术,膀胱丰满不能在测试开始前评估。此外,排尿的变化可能由于行为,而不是疾病进展的直?…
Divulgations
The authors have nothing to disclose.
Acknowledgements
我们要感谢艾米莉·里克、克里斯汀·乌赫特曼和里克实验室在畜牧业方面的协助和对这份手稿的反馈。我们要感谢NIDDK和NIEHS对这些研究的财政支持:U54 DK104310(WAR、JAM、PCM、CMV、DEB)、R01 ES001332(WAR、CMV)、K12 DK100022(TTL、AR-A、DH)。内容由作者全权负责,并不代表国家卫生研究院的官方观点。
Materials
| 21mm Clear Tubing | Supera Anesthesia Innov | 301-150 | |
| 27 gauge needle | BD | Z192376 | |
| 4 port Manifold | Supera Anesthesia Innov | RES536 | |
| DEFINITY | Lantheus Medical Imaging | DE4 | |
| F/AIR Canister | Supera Anesthesia Innov | 80120 | |
| Graefe forceps (Serrated, Straight) | F.S.T. | 11050-10 | |
| Inlet/Outlet Fittings | Supera Anesthesia Innov | VAP203/4 | |
| Isoflurane | Midwest Vet Supply | 193.33161.3 | |
| Isoflurane Vaporizer | Supera Anesthesia Innov | VAP3000 | |
| MV707 probe | Fujifilm VisualSonics Inc | ||
| Oxygen Flowmeter | Supera Anesthesia Innov | OXY660 | |
| Polyethylene 50 tubing | BD | 427516 | |
| Pressure Reg/Gauge | Supera Anesthesia Innov | OXY508 | |
| Rebreathing Circuits | Supera Anesthesia Innov | CIR529 | |
| Small Mice Nose Cone | Supera Anesthesia Inov | ACC526 | |
| Sterile saline | Midwest Vet Supply | 193.74504.3 | NaCl 0.9%, Injectable |
| Straight Sharp/Blunt Scissors | Fine Scientific Tools (F.S.T) | 14054-13 | |
| Syringe | BD | 309646 | 5mL |
| Vevo 770 | Fujifilm VisualSonics Inc | ||
| VIALMIX | Lantheus Medical Imaging | VMIX |
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