Nunnari J J, Zand T, Joris I, Majno G
Department of Pathology, University of Massachusetts Medical School, Worcester 01655.
Exp Mol Pathol. 1989 Aug;51(1):1-8. doi: 10.1016/0014-4800(89)90002-6.
A technique is described which provides morphologic and quantitative data on the amount of oil red O (ORO) staining in thoracic aortas of rats fed a high cholesterol diet. Samples are stained with ORO, the dye is extracted, and the concentration of ORO in the extract is measured colorimetrically. Wistar rats fed ad libitum either standard chow (control group: n = 15) or chow supplemented with 4% cholesterol, 1% cholic acid, and 0.5% thiouracil (CCT group: n = 23) were maintained on these diets for 1, 3, 6, 9, or 12 months. Plasma cholesterol levels averaged overall 87 and 737 mg/dl for the control and CCT groups, respectively. Animals were killed under anesthesia by perfusion fixation with formalin or glutaraldehyde, and samples of thoracic aorta were stained with ORO. After microscopic study en face and measurement of surface area, the ORO was extracted in chloroform-methanol (2:1). Concentrations of ORO (microM) were determined from a standard curve and expressed as microM/mm2 of aorta. Aortas of CCT animals showed progressive diet- and time-dependent increases in the amount of ORO staining compared to controls. We conclude that this method yields reliable quantitative data applicable to studying atherosclerosis in small animals.
本文描述了一种技术，该技术可提供喂食高胆固醇饮食大鼠胸主动脉中油红O（ORO）染色量的形态学和定量数据。样本用ORO染色，提取染料，并通过比色法测量提取物中ORO的浓度。随意喂食标准饲料（对照组：n = 15）或添加4%胆固醇、1%胆酸和0.5%硫脲的饲料（CCT组：n = 23）的Wistar大鼠维持这些饮食1、3、6、9或12个月。对照组和CCT组的血浆胆固醇水平总体平均分别为87和737 mg/dl。动物在麻醉下通过用福尔马林或戊二醛灌注固定处死，胸主动脉样本用ORO染色。在进行表面观察和表面积测量的显微镜研究后，用氯仿 - 甲醇（2:1）提取ORO。根据标准曲线确定ORO的浓度（微摩尔），并表示为每平方毫米主动脉的微摩尔数。与对照组相比，CCT组动物的主动脉中ORO染色量呈现出饮食和时间依赖性的逐渐增加。我们得出结论，该方法可产生适用于研究小动物动脉粥样硬化的可靠定量数据。
