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对L-精氨酸甲酯(L-NAME)的渗漏反应因用于标记白蛋白的荧光染料而异。

Leakage responses to L-NAME differ with the fluorescent dye used to label albumin.

作者信息

Rumbaut R E, Harris N R, Sial A J, Huxley V H, Granger D N

机构信息

Department of Internal Medicine, Pulmonary and Critical Care Division, and Department of Physiology, University of Missouri-Columbia, Columbia, Missouri 65212, USA.

出版信息

Am J Physiol. 1999 Jan;276(1):H333-9. doi: 10.1152/ajpheart.1999.276.1.H333.

Abstract

Nitric oxide synthase (NOS) inhibitors have been reported to increase as well as to decrease microvascular transport of macromolecules in a variety of models. This study was performed to determine whether the influence of NOS inhibition on albumin leakage was dependent on the fluorescent dyes used to label albumin. Albumin leakage was assessed in rat mesenteric venules during control conditions and after exposure to the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME). Albumin was labeled with any one of four dyes: FITC, sulforhodamine 101 [Texas Red (TR)], dichlorotriazinyl aminofluorescein (DTAF), or Oregon Green 514 (OG). Superfusion with L-NAME (10(-4) M) was accompanied by an increase in leakage of FITC-labeled albumin (n = 12) but not of albumin labeled with DTAF (n = 10), TR (n = 10), or OG (n = 4). In vessels perfused with both FITC- and TR-labeled albumin (n = 12), superfusion with L-NAME increased leakage of FITC- but not TR-labeled albumin. In conclusion, albumin leakage responses to L-NAME differ among various fluorescent dyes. Therefore, caution is advised in comparison of albumin leakage results that utilize different fluorescent dyes.

摘要

据报道,在多种模型中,一氧化氮合酶(NOS)抑制剂既能增加也能减少大分子的微血管转运。本研究旨在确定NOS抑制对白蛋白渗漏的影响是否取决于用于标记白蛋白的荧光染料。在对照条件下以及暴露于NOS抑制剂NG-硝基-L-精氨酸甲酯(L-NAME)后,评估大鼠肠系膜小静脉中的白蛋白渗漏情况。白蛋白用四种染料中的任何一种进行标记:异硫氰酸荧光素(FITC)、磺基罗丹明101[德克萨斯红(TR)]、二氯三嗪基氨基荧光素(DTAF)或俄勒冈绿514(OG)。用L-NAME(10^(-4) M)进行灌流时,FITC标记的白蛋白渗漏增加(n = 12),但DTAF标记的白蛋白(n = 10)、TR标记的白蛋白(n = 10)或OG标记的白蛋白(n = 4)渗漏未增加。在用FITC和TR标记的白蛋白进行灌流的血管中(n = 12),用L-NAME进行灌流会增加FITC标记的白蛋白渗漏,但不会增加TR标记的白蛋白渗漏。总之,不同荧光染料对L-NAME的白蛋白渗漏反应不同。因此,在比较使用不同荧光染料的白蛋白渗漏结果时应谨慎。

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