Measurement of plasma volume in rats with use of fluorescent-labeled albumin molecules.
Measurement of plasma volume in rats with use of fluorescent-labeled albumin molecules.
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DOI：
10.1152/jappl.1994.76.1.485
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发表时间：
1994
影响因子：
3.3
通讯作者：
C. Gillen;A. Takamata;G. Mack;E. Nadel
中科院分区：
文献类型：
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作者：
C. Gillen;A. Takamata;G. Mack;E. Nadel
我们描述了一种在小动物体内测量等离子体体积（PV）的方法，该方法允许小样本量，但不需要使用放射性同位素，因此是一种方便的重复测量方法。德克萨斯红共价结合白蛋白（TR-A）用于典型的指标稀释技术来测量PV。TR-A在激发λ为590 nm，发射λ为610 nm时的相对荧光强度与其浓度（最高为0.15 mg/ml）呈线性关系。将导管经麻醉大鼠右颈静脉插入腔静脉。取0.5 ml对照血，注射一定量的TR-A，并用生理盐水冲洗导管。注射TR-A后5分钟取0.5 ml注射后标本。通过比较对照和注射后血浆样品相对荧光强度与标准样品的差异来计算PV。22只大鼠的PV值[362 +/- 14 (SE) g]分别为：TR-A法测14.1 +/- 0.4 ml （39.6 +/- 0.9 ml/kg体wt）和标准放射性碘化白蛋白法测12.8 +/- 0.4 ml （35.9 +/- 1.0 ml/kg体wt）。两种方法测得的同一大鼠PV值有较强的相关性（r = 0.90, P < 0.01）。注射实验表明，TR-A方法可以检测急性PV变化，并且在长期注射的大鼠身上重复测量PV，表明该方法可以连续几天可靠地测量PV。
We describe a method for measuring plasma volume (PV) in small animals that allows small sample sizes but does not require the use of radioisotopes and thus is a convenient approach for making repeated measurements. Texas Red covalently bound to albumin (TR-A) was used in a typical indicator-dilution technique to measure PV. The relative fluorescent intensity of TR-A is linear to its concentration (up to 0.15 mg/ml) at an excitation lambda of 590 nm and an emission lambda of 610 nm. Catheters were inserted through the right jugular vein of anesthetized rats and threaded into the vena cava. A 0.5-ml control blood sample was taken, a measured quantity of TR-A was injected, and the catheter was flushed with saline. A 0.5-ml postinjection sample was taken 5 min after TR-A injection. PV was calculated by comparing the difference between the relative fluorescent intensity of control and postinjection plasma samples to a standard. The PV of 22 rats [362 +/- 14 (SE) g] was 14.1 +/- 0.4 ml (39.6 +/- 0.9 ml/kg body wt) measured by the TR-A method and 12.8 +/- 0.4 ml (35.9 +/- 1.0 ml/kg body wt) measured by a standard radioiodinated albumin method. There was a strong correlation between PV measured by both methods in the same rat (r = 0.90, P < 0.01). Infusion experiments indicated that the TR-A method can detect acute changes in PV, and repeated measurements of PV made on a chronically instrumented rat demonstrated that the method can reliably measure PV on consecutive days.