小鼠主动脉斑块大体油红O染色技术的改进

Improvement of gross oil red O staining technique for aortic plaque in mice

目的对小鼠主动脉斑块大体油红O染色技术进行改进,以准确测定动脉粥样硬化小鼠主动脉壁斑块面积。方法选取普通饲料喂养10月龄雄性ApoE-/-小鼠行整根主动脉剥离,显微弹簧剪去除主动脉外膜脂肪并纵向剖开整根主动脉,不同温度梯度下(25℃、45℃、65℃)油红O染料孵育10 min,吸水纸上铺展主动脉,用图像分析软件测定主动脉各部位的斑块面积。结果不同温度油红O染色发现,45℃较25℃孵育动脉斑块着色明显,呈血红色,易于测量,而65℃孵育条件下,斑块出现脱落现象,故45℃为最佳温度。油红O染色显示ApoE-/-小鼠的主动脉弓、胸主动脉、腹主动脉以及髂总动脉均可见明显的粥样硬化斑块,大体主动脉斑块面积为(29.8±3.2)%;主动脉弓部的斑块面积为(64.4±5.4)%,与其他部位的斑块面积相比较,弓部更易发生病变,是其他部位的2~4倍。结论全面改进小鼠主动脉斑块大体油红O染色技术并准确测量小鼠大体及各部位主动脉斑块面积,为评估动脉粥样硬化病变程度提供技术指导。

Objective The gross oil red O staining technique for aortic plaque in mice was improved to accurately evaluate the aortic plaque area in atherosclerotic mice.Methods Aortic dissection was performed on male ApoE-/-mice at the age of 10 months with normal-diet.Microspring scissors were used to remove adventitial fat and to cut longitudinally the entire aorta.The aorta was incubated with oil red O dye in different temperature conditions(25℃,45℃and 65℃)for 10 minutes,and then the gross aortawas spread out on the absorbent paper.Photograph was capturedand the plaque area of each part was measured by Image J software.Results Compared with the aortic oil red O staining under the incubation condition at 25℃,the aortic plaques were much more stained than that at 45℃,which showed deep red color and easy for measure.While under the condition of 65℃,plaque shed.Thus,45℃was defined as the optimal tem-perature.Oil red O staining showed significant atherosclerotic plaques in the aortic arch,thoracic aorta,abdominal aorta,and biforked common iliac artery of ApoE-/-miceand the gross aortic plaque size was(29.8±3.2)%.While,the plaque area of the aortic arch was(64.4±5.4)%,whichwas 2-4 times than that of the other parts,indicating that the aortic arch was more prone to lesions than other parts.Conclusion The gross oil red O staining technique for aortic plaque in mice was comprehensively improved and the area of gross and local aortic plaque could be accurately and easily measured,providing a certain technical guidance to evaluate the degree of atherosclerotic lesions.