Ferrero Maria Elena
Institute of General Pathology, University of Milan, Milan, Italy.
Methods Mol Med. 2004;98:191-8. doi: 10.1385/1-59259-771-8:191.
This chapter describes the methods for measuring the increase in vascular permeability induced by tumor necrosis factor alpha (TNFalpha, or other mediators, in vivo in animal models. Mouse liver or kidney are perfused through the portal vein or the renal artery, respectively, by intravascular injection of a blue dye (trypan blue or Evans blue) combined with albumin. When endothelial permeability is increased, the dye extravasates and reaches the subendothelial spaces. Thus, subsequent washing of dye-perfused organs with saline cannot remove the dye. The perfusates are drawn from the suprahepatic inferior vena cava (for the liver) and from the renal vein (for the kidney). After perfusion, the livers and kidneys are removed, homogenized, and centrifuged. Spectrophotometric analysis of supernatants, at 540 nm, is then performed. The increase in optical density values is indicative of the increase in dye retention, hence of vascular leakage.
本章介绍了在动物模型体内测量肿瘤坏死因子α（TNFα）或其他介质诱导的血管通透性增加的方法。分别通过门静脉或肾动脉向小鼠肝脏或肾脏灌注血管内注射蓝色染料（台盼蓝或伊文思蓝）与白蛋白的混合物。当内皮通透性增加时，染料渗出并到达内皮下间隙。因此，随后用生理盐水冲洗染料灌注的器官无法去除染料。灌注液分别从肝上腔静脉（用于肝脏）和肾静脉（用于肾脏）抽取。灌注后，取出肝脏和肾脏，匀浆并离心。然后对上清液在540nm处进行分光光度分析。光密度值的增加表明染料潴留增加，即血管渗漏增加。
