Sasaki Yui, Iwama Ryosuke, Sato Tsubasa, Heishima Kazuki, Shimamura Shunsuke, Ichijo Tosihiro, Satoh Hiroshi, Furuhama Kazuhisa
Cooperative Department of Veterinary Medicine, Iwate University, Morioka, Iwate, Japan.
United Graduate School of Veterinary Science, Gifu University, Gifu, Japan.
Physiol Rep. 2014 Aug 28;2(8). doi: 10.14814/phy2.12135. Print 2014 Aug 1.
To develop an expedient procedure for estimating glomerular filtration rate (GFR) in conscious mice, we first established a simple technique for repeated blood collections from the tail vein using a microcapillary tube attached to a 24-gauge needle without the hub. Then, we devised a definition equation for estimation of the GFR using the contrast medium iodixanol as a test tracer. Iodixanol was administered as a bolus injection at 1500 mg I/kg to ddY mice, and the GFR was determined by the conventional multisample strategy. Based on cumulative data from the multisample method, an equation for the single-blood-sample method including the iodixanol dose, estimated distribution volume (Vd), and plasma iodixanol concentration at 60 min later was sought. The GFR values from the multisample method were in good agreement with those calculated using the equation. In clinically healthy mice, the GFR decreased gradually with ages from 11 weeks old in both sexes, suggesting the necessity of the corresponding control in each protocol. In nephropathy mice induced by cisplatin, the GFR values decreased with rises in serum BUN and creatinine concentrations, and serum creatinine became elevated when the GFR decreased to approximately 70% of the basal value. The results suggest that the simplified equation using iodixanol, allowing for the repeated use of the same mice, is a versatile procedure for research purposes.
为了开发一种在清醒小鼠中估算肾小球滤过率(GFR)的便捷方法,我们首先建立了一种简单的技术,使用连接到无针座的24号针头的微量毛细管从尾静脉重复采集血液。然后,我们设计了一个使用造影剂碘克沙醇作为测试示踪剂估算GFR的定义方程。将碘克沙醇以1500 mg I/kg的剂量推注给ddY小鼠,并通过传统的多样本策略测定GFR。基于多样本方法的累积数据,寻找一个单血样方法的方程,该方程包括碘克沙醇剂量、估算分布容积(Vd)和60分钟后的血浆碘克沙醇浓度。多样本方法得到的GFR值与使用该方程计算的值高度一致。在临床健康的小鼠中,两性的GFR从11周龄开始随年龄逐渐下降,这表明在每个实验方案中都需要进行相应的对照。在顺铂诱导的肾病小鼠中,GFR值随血清尿素氮和肌酐浓度的升高而降低,当GFR降至基础值的约70%时,血清肌酐升高。结果表明,使用碘克沙醇的简化方程允许对同一只小鼠进行重复使用,是一种适用于研究目的的通用方法。