Schock-Kusch Daniel, Sadick Maliha, Henninger Nadja, Kraenzlin Bettina, Claus Guenter, Kloetzer Hans-Martin, Weiss Christel, Pill Johannes, Gretz Norbert
InnovationLab GmbH, Speyerer Str. 4, D-69115 Heidelberg, Germany.
Nephrol Dial Transplant. 2009 Oct;24(10):2997-3001. doi: 10.1093/ndt/gfp225. Epub 2009 May 21.
Inulin/sinistrin (I/S) clearance is a gold standard for an accurate assessment of glomerular filtration rate (GFR). Here we describe and validate an approach for a transcutaneous determination of GFR by using fluorescein-isothiocyanate-labelled sinistrin (FITC-S) in rats.
Using a small animal imager, fluorescence is measured over the depilated ear of a rat after the injection of FITC-S. The decay curve of fluorescence is used for the calculation of half-life and GFR. The thus obtained transcutaneous data were validated by simultaneously performed enzymatic and fluorometric measurements in plasma of both FITC-S and sinistrin.
The results of enzymatic sinistrin determination versus transcutaneous half-life of FITC-S or plasma fluorescence correlated well with each other (R(2) > 0.90). Furthermore, Bland-Altman analyses proved a good degree of agreement of the three methods used. The measurements performed in healthy animals as well as different models of renal failure demonstrate its appropriateness in a wide range of renal function.
The transcutaneous method described offers a precise assessment of GFR in small animals. As neither blood and/or urine sampling nor time-consuming lab work is required, GFR can be determined immediately after the clearance procedure is finished. This method, therefore, simplifies and fastens GFR determinations in small lab animals compared to conventional bolus clearance techniques based on blood sampling. A low-cost device for the measurement of transcutaneous fluorescence intensity over time is under construction.
菊粉/左聚糖(I/S)清除率是准确评估肾小球滤过率(GFR)的金标准。在此,我们描述并验证了一种通过使用异硫氰酸荧光素标记的左聚糖(FITC-S)经皮测定大鼠GFR的方法。
使用小动物成像仪,在注射FITC-S后测量大鼠脱毛耳部的荧光。荧光衰减曲线用于计算半衰期和GFR。通过同时对FITC-S和左聚糖血浆进行酶法和荧光法测量,对由此获得的经皮数据进行验证。
酶法测定左聚糖与FITC-S的经皮半衰期或血浆荧光结果之间相关性良好(R²>0.90)。此外,Bland-Altman分析证明所使用的三种方法具有良好的一致性。在健康动物以及不同肾衰竭模型中进行的测量表明其在广泛的肾功能范围内具有适用性。
所描述的经皮方法可对小动物的GFR进行精确评估。由于既不需要采集血液和/或尿液样本,也不需要耗时的实验室工作,因此在清除程序完成后即可立即测定GFR。因此,与基于采血的传统推注清除技术相比,该方法简化并加快了小型实验动物GFR的测定。一种用于随时间测量经皮荧光强度的低成本设备正在研制中。