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首页> 外文期刊>American Journal of Veterinary Research >Measurement of plasma renin concentration in cats by use of a fluorescence resonance energy transfer peptide substrate of renin
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Measurement of plasma renin concentration in cats by use of a fluorescence resonance energy transfer peptide substrate of renin

机译：利用肾素的荧光共振能量转移肽底物测量猫血浆中肾素的浓度

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Objective-To evaluate the use of a commercially available 5-carboxyfluorescein-based, intramolecularly quenched, fluorescence resonance energy transfer (FRET) peptide substrate of renin for measurement of plasma renin concentration in cats.Sample Population-Plasma samples obtained during a previous study of renal autograft ischemia-reperfusion injury in 10 cats and samples of fetal bovine serum containing recombinant human renin (rh-renin).Procedures-Experiments involving samples of fetal bovine serum containing rh-renin were conducted to identify a suitable control vehicle, optimal substrate concentration, and appropriate duration of incubation. With the use of the identified assay conditions, a Standard curve was constructed to allow conversion of relative fluorescent units into values of renin concentration (ng/mL). Subsequently, plasma samples obtained from cats before and after renal autograft ischemia-reperfusion injury were assayed to determine endogenous renin concentration.Results-Under conditions of a 1:50 substrate dilution and 4-hour incubation period, the assay detected small amounts of rh-renin in fetal bovine serum. A linear relationship (R-2 = 0.996) between the relative fluorescent units generated and exogenous rh-renin concentration was evident. The assay detected renin in plasma samples obtained from cats after renal autograft ischemia-reperfusion, and renin concentrations on days 1 and 2 after transplant differed significantly.Conclusions and Clinical Relevance-The study data indicated that the assay involving the FRET peptide substrate of renin is potentially a rapid and specific method for measurement of plasma renin concentration in cats. (Am J Vet Res 2009;70:1315-1322)
机译：目的-评估市售的基于5羧基荧光素的分子内淬灭的肾素荧光共振能量转移（FRET）肽底物在猫血浆血浆中肾素浓度的测量中的用途。 10只猫的肾脏自体移植缺血再灌注损伤和包含重组人肾素（rh-renin）的胎牛血清样品。进行了涉及含rh-renin胎牛血清样品的程序-实验，以确定合适的对照载体，最佳底物浓度，以及适当的孵育时间。利用确定的测定条件，构建标准曲线以允许将相对荧光单位转换为肾素浓度（ng / mL）的值。随后，对从猫自体肾缺血再灌注损伤前后的血浆样品进行测定，以确定内源性肾素浓度。结果-在1:50底物稀释度和4小时孵育时间的条件下，该检测方法检测到少量的rh-胎牛血清中的肾素。产生的相对荧光单位与外源性rh-肾素浓度之间存在线性关系（R-2 = 0.996）。该方法检测的猫自体肾脏缺血再灌注后血浆样品中的肾素含量，以及移植后第1天和第2天的肾素浓度存在显着差异。结论和临床意义-研究数据表明，涉及肾素FRET肽底物的检测方法为可能是一种快速而具体的方法来测量猫血浆中肾素浓度。 （Am J Vet Res 2009; 70：1315-1322）

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