Beattie John H, Duthie Susan J, Kwun In-Sook, Ha Tae-Youl, Gordon Margaret-Jane
Division of Vascular Health, Rowett Institute of Nutrition and Health, University of Aberdeen, Aberdeen, UK.
J Vasc Res. 2009;46(4):347-52. doi: 10.1159/000189795. Epub 2009 Jan 10.
The quantification of aortic lesions is an important end-point analysis for evaluating atherogenesis in mouse models of atherosclerosis. Morphometric methods involving the staining of aorta with a Sudan lysochrome followed by image analysis of the stained lesion area are commonly used. We have developed a more rapid method involving solubilisation of the stain retained by aortic lesions. In 2 separate studies, 5-week-old male apoE(-/-) and C57BL/6 wild-type (apoE(+/+)) mice were given a high fat (21%), Western-type diet for 13, 15 or 25 weeks. At study termination, the descending thoracic aorta (DA) and/or aortic arch (AA) were stained with Oil Red O (ORO). The incorporated stain was extracted using chloroform/methanol (2:1) solvent and quantified by spectrophotometry at 520 nm. In study 1 (13 weeks), ORO stain in the AA and DA of apoE(-/-) mice was 1.9 and 1.4 times higher than background staining of apoE(+/+) aorta tissue, respectively. At 15 and 25 weeks (study 2), ORO stain in the AA of apoE(-/-) mice was 1.9 and 2.5 times higher than the background, respectively. We conclude that the ORO solubilisation technique applied to AA samples is a very useful and rapid method for atherosclerotic lesion quantification.
主动脉病变的量化是评估动脉粥样硬化小鼠模型中动脉粥样硬化形成的重要终点分析。常用的形态测量方法包括用苏丹溶色素对主动脉进行染色，然后对染色的病变区域进行图像分析。我们开发了一种更快的方法，涉及溶解主动脉病变保留的染料。在两项独立研究中，给5周龄雄性载脂蛋白E基因敲除（apoE(-/-)）和C57BL/6野生型（apoE(+/+)）小鼠喂食高脂肪（21%）的西式饮食13、15或25周。在研究结束时，用油红O（ORO）对胸降主动脉（DA）和/或主动脉弓（AA）进行染色。使用氯仿/甲醇（2:1）溶剂提取掺入的染料，并通过在520nm处的分光光度法进行定量。在研究1（13周）中，apoE(-/-)小鼠AA和DA中的ORO染色分别比apoE(+/+)主动脉组织的背景染色高1.9倍和1.4倍。在15周和25周（研究2）时，apoE(-/-)小鼠AA中的ORO染色分别比背景高1.9倍和2.5倍。我们得出结论，应用于AA样本的ORO溶解技术是一种非常有用且快速的动脉粥样硬化病变量化方法。
