Practical assessment of the quantification of atherosclerotic lesions in apoE-/- mice
Practical assessment of the quantification of atherosclerotic lesions in apoE-/- mice
复制标题
DOI：
10.3892/mmr.2015.4084
复制
发表时间：
2015-10-01
影响因子：
3.4
通讯作者：
Liu, Enqi
中科院分区：
文献类型：
--
作者：
Lin, Yan;Bai, Liang;Liu, Enqi
基因操作使小鼠能够被广泛用作研究人类动脉粥样硬化疾病机制的动物模型。然而，在不同的实验室中没有标准的方法来量化动脉粥样硬化病变。本研究介绍了一个全面和精确的定量评估小鼠动脉粥样硬化病变。在本研究中，6周龄apoE（-/-）小鼠喂食普通饲料或高脂饲料（HFD）12周。每四周测量一次血脂水平。使用图像分析系统对主动脉粥样硬化病变进行定量和分析。分离主动脉树并用油红O染色以测量大体病变面积。取出心脏并分成连续的横截面，然后如下评估主动脉根部的显微内膜病变：（1）进行弹性货车Gieson染色以确定动脉粥样硬化病变的面积;（2）横截面用苏木精和伊红染色以进行组织学分析;（3）切片行油红0染色和免疫组化染色，对病变内细胞成分进行定量分析。喂食普通饲料或HFD的ApoE小鼠在主动脉根部发生严重的动脉粥样硬化，然而，在主动脉树的其余部分几乎没有病变。与对照组相比，HFD apoE-/-小鼠的血浆脂质水平升高，主动脉树的大体病变面积、主动脉根部的显微病变面积以及病变内存在的巨噬细胞、血管平滑肌细胞和中性脂质的数量增加。ApoE（-/-）小鼠喂食HFD可加速动脉粥样硬化的发展。在本研究中描述的定量方法可用于协助在小鼠动脉粥样硬化的未来调查。
Genetic manipulations have enabled the mouse to be widely used as an animal model for investigating the mechanisms of human atherosclerotic disease. However, there is no standard method for quantifying atherosclerotic lesions among different laboratories. The present study introduces a thorough and precise quantitative assessment of atherosclerotic lesions in mice. In the present study, 6-week-old apoE(-/-) mice were fed either a chow diet or a high-fat diet (HFD) for 12 weeks. Plasma lipid levels were measured every four weeks. Aortic atherosclerotic lesions were quantified and analyzed using an image analysis system. The aortic tree was isolated and stained with Oil Red O to measure the gross lesion area. The heart was removed and divided into sequential cross sections, which were then assessed for microscopic intimal lesions in the aortic root as follows: (1) Elastic van Gieson staining was performed to determine the area of the atherosclerotic lesion; (2) cross sections were stained with hematoxylin and eosin for histological analysis; and (3) cross sections were stained with Oil Red 0 and immunohistochemical staining for quantitative analysis of the cellular components within the lesions. ApoE mice fed with either the chow diet or HFD developed severe atherosclerosis in the aortic root, however, there were few lesions in the remainder of the aortic tree. Compared with the control group, the HFD apoE-/- mice had increased plasma lipid levels and increases in the gross lesion area in the aortic tree, the microscopic lesion area in the aortic root and the number of macrophages, vascular smooth muscle cells and neutral lipids present within the lesions. HFD feeding in the apoE(-/-) mice accelerated the development of atherosclerosis. The quantitative method described in the present study may be used to assist in future investigations of atherosclerosis in mice.