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建立尿斑实验系统用于小鼠下尿路功能检测

罗闯, 杨佳丽, 刘娟, 邱湘, 谢翔, 陈环^*

西南医科大学基础医学院，泸州 646000

摘要

本文旨在提供一套简单且可靠的尿斑实验系统用于检测小鼠下尿路功能，促进尿斑实验标准化。本系统利用紫外光对尿斑进行可视化，筛选出没有自发荧光且对尿液具有良好扩散作用的滤纸，比较不同波长紫外光对尿斑成像效果。制作尿液体积-尿斑面积标准曲线，明确所选滤纸上形成的尿斑面积和尿液体积的关系，利用建立的尿斑实验系统对12~13周龄C57BL/6J雌性小鼠进行排尿模式分析，并与其他实验室采用相同实验参数设置的尿斑实验结果进行比较。最后利用该系统跟踪尿路感染小鼠和运输应激小鼠排尿模式变化。结果显示：#4滤纸无自发荧光且对尿液具有较好扩散作用，在365 nm紫外光照射下该滤纸上尿斑成像清晰且边界清楚，在尿斑实验中0.7 mm厚度#4号滤纸能保持相对较好的完整性；在该滤纸上，尿液形成的尿斑面积和尿液体积成线性关系；采用该尿斑实验系统检测野生C57BL/6J雌性小鼠4 h尿量约为695.8 μL，主尿斑平均体积约为126.4 μL，超过84%主尿斑体积在20~200 μL，主尿斑数约为5.5个，不同实验室小鼠在主尿斑体积上具有一致性；通过该系统检测小鼠在尿路感染或运输应激后排尿次数增多，主尿斑体积减少，主尿斑数量增加。综上，本研究提供了一套简单且可靠的尿斑实验系统，能对正常小鼠、尿路感染小鼠和运输应激小鼠的排尿模式进行跟踪，检测其下尿路功能变化。

关键词： 尿斑实验; 排尿模式; 尿路感染; 主尿斑; 运输应激

Evaluation of the lower urinary tract function using voiding spot assay in mice

LUO Chuang, YANG Jia-Li, LIU Juan, QIU Xiang, XIE Xiang, CHEN Huan^*

School of Basic Medical Sciences, Southwest Medical University, Luzhou 646000, China

Abstract

The objective of present study was to develop a simple and reliable voiding spot assay (VSA) system to evaluate the lower urinary tract function of mice, and to establish it as a standardized protocol. Ultraviolet (UV) light was used to screen out the filter paper without autofluorescence and with optimal urine diffusion properties. Next, the appropriate wavelength of UV was determined based on the quality of the photographic image of urine spots on the filter paper. To confirm that the urine stain area on the fliter paper was correlated with the amount of urine, a volume-area standard curve was constructed. The utility of this VSA system was validated using female wild-type C57BL/6J mice aged 12–13 weeks, and the data generated under identical procedural settings were compared among laboratories. Furthermore, this VSA system was employed to analyze the changes in voiding patterns in mice with urinary tract infections or transportation stress. No. 4 filter paper with a thickness of 0.7 mm was identified as the most suitable material for VSA, exhibiting no autofluorescence and facilitating optimal urine diffusion. The filter paper retained its integrity during the assay, and there was a linear correlation between urine volume and stained area under 365 nm UV light. Utilizing this VSA system, we determined that female wild-type C57BL/6J mice produced approximately 695.8 μL total urine and 5.5 primary voiding spots (PVS) with an average size of 126.4 μL/spot within 4-h period. Over 84% of PVS volumes ranged from 20 to 200 μL. Notably, PVS volumes of mice were similar across different laboratories. Mice with urinary tract infections or transportation stress exhibited significant changes in VSA parameters, including increased voiding frequency, PVS number, and decreased PVS volume. Therefore, this VSA system can be used to evaluate the urinary function of normal mice, as well as those with urinary tract infection or transportation stress.

Key words: voiding spot assay; voiding pattern; urinary tract infection; primary voiding spot; transportation stress

收稿日期：　　录用日期：

通讯作者：陈环　　E-mail: huanchen@swmu.edu.cn

DOI: 10.13294/j.aps.2023.0049

引用本文：

罗闯, 杨佳丽, 刘娟, 邱湘, 谢翔, 陈环. 建立尿斑实验系统用于小鼠下尿路功能检测[J]. 生理学报 2023; 75 (5): 636-646.

LUO Chuang, YANG Jia-Li, LIU Juan, QIU Xiang, XIE Xiang, CHEN Huan. Evaluation of the lower urinary tract function using voiding spot assay in mice. Acta Physiol Sin 2023; 75 (5): 636-646 (in Chinese with English abstract).