Practical assessment of the quantification of atherosclerotic lesions in apoE‑/‑ mice

Lin,Yan; Bai,Liang; Chen,Yulong; Zhu,Ninghong; Bai,Yanping; Li,Qianwei; Zhao,Sihai; Fan,Jianglin; Liu,Enqi

doi:10.3892/mmr.2015.4084

Practical assessment of the quantification of atherosclerotic lesions in apoE‑/‑ mice

Authors:
- Yan Lin
- Liang Bai
- Yulong Chen
- Ninghong Zhu
- Yanping Bai
- Qianwei Li
- Sihai Zhao
- Jianglin Fan
- Enqi Liu
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Affiliations: Laboratory Animal Center, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China, Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Institute of Basic and Translational Medicine, Xi'an Medical University, Xi'an, Shaanxi 710021, P.R. China, Department of Molecular Pathology, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Yamanashi 409‑3898, Japan
Published online on: July 16, 2015 https://doi.org/10.3892/mmr.2015.4084
Pages: 5298-5306

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Abstract

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 O 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.

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1	Daugherty A: Mouse models of atherosclerosis. Am J Med Sci. 323:3–10. 2002. View Article : Google Scholar : PubMed/NCBI
2	Kapourchali FR, Surendiran G, Chen L, Uitz E, Bahadori B and Moghadasian MH: Animal models of atherosclerosis. World J Clin Cases. 2:126–132. 2014. View Article : Google Scholar : PubMed/NCBI
3	Zaragoza C, Gomez-Guerrero C, Martin-Ventura JL, Blanco-Colio L, Lavin B, Mallavia B, Tarin C, Mas S, Ortiz A and Egido J: Animal models of cardiovascular diseases. J Biomed Biotechnol. 2011:4978412011. View Article : Google Scholar : PubMed/NCBI
4	Knowles JW and Maeda N: Genetic modifiers of atherosclerosis in mice. Arterioscler Thromb Vasc Biol. 20:2336–2345. 2000. View Article : Google Scholar : PubMed/NCBI
5	Plump AS, Smith JD, Hayek T, Aalto-Setälä K, Walsh A, Verstuyft JG, Rubin EM and Breslow JL: Severe hypercholes-terolemia and atherosclerosis in apolipoprotein E-deficient mice created by homologous recombination in ES cells. Cell. 71:343–353. 1992. View Article : Google Scholar : PubMed/NCBI
6	Zadelaar S, Kleemann R, Verschuren L, de Vries-Van der Weij J, van der Hoorn J, Princen HM and Kooistra T: Mouse models for atherosclerosis and pharmaceutical modifiers. Arterioscler Thromb Vasc Biol. 27:1706–1721. 2007. View Article : Google Scholar : PubMed/NCBI
7	Daugherty A, Lu H, Howatt DA and Rateri DL: Modes of defining atherosclerosis in mouse models: Relative merits and evolving standards. Methods Mol Biol. 573:1–15. 2009. View Article : Google Scholar : PubMed/NCBI
8	Nakashima Y, Plump AS, Raines EW, Breslow JL and Ross R: ApoE-deficient mice develop lesions of all phases of athero sclerosis throughout the arterial tree. Arterioscler Thromb. 14:133–140. 1994. View Article : Google Scholar : PubMed/NCBI
9	Mani S, Li H, Untereiner A, Wu L, Yang G, Austin RC, Dickhout JG, Lhoták Š, Meng QH and Wang R: Decreased endogenous production of hydrogen sulfide accelerates atherosclerosis. Circulation. 127:2523–2534. 2013. View Article : Google Scholar : PubMed/NCBI
10	Wang Y, Zhao X, Jin H, Wei H, Li W, Bu D, Tang X, Ren Y, Tang C and Du J: Role of hydrogen sulfide in the development of athero-sclerotic lesions in apolipoprotein E knockout mice. Arterioscler Thromb Vasc Biol. 29:173–179. 2009. View Article : Google Scholar
11	Paigen B, Morrow A, Holmes PA, Mitchell D and Williams RA: Quantitative assessment of atherosclerotic lesions in mice. Atherosclerosis. 68:231–240. 1987. View Article : Google Scholar : PubMed/NCBI
12	Purcell-Huynh DA, Farese RV Jr, Johnson DF, Flynn LM, Pierotti V, Newland DL, Linton MF, Sanan DA and Young SG: Transgenic mice expressing high levels of human apolipoprotein B develop severe atherosclerotic lesions in response to a high-fat diet. J Clin Invest. 95:2246–2257. 1995. View Article : Google Scholar : PubMed/NCBI
13	Véniant MM, Pierotti V, Newland D, Cham CM, Sanan DA, Walzem RL and Young SG: Susceptibility to atherosclerosis in mice expressing exclusively apolipoprotein B48 or apolipoprotein B100. J Clin Invest. 100:180–188. 1997. View Article : Google Scholar : PubMed/NCBI
14	Tangirala RK, Rubin EM and Palinski W: Quantitation of atheroscleros in murine models: Correlation between lesions in the aortic origin and in the entire aorta and differences in the extent of lesions between sexes in LDL receptor-deficient and apolipoprotein E-deficient mice. J Lipid Res. 36:2320–2328. 1995.PubMed/NCBI
15	Guide for the Care and Use of Laboratory Animals. US National Institutes of Health; Bethesda: 2011
16	Chen Y, Zhao S, Huang B, Wang Y, Li Y, Waqar AB, Liu R, Bai L, Fan J and Liu E: Probucol and cilostazol exert a combinatorial anti-atherogenic effect in cholesterol-fed rabbits. Thromb Res. 132:565–571. 2013. View Article : Google Scholar : PubMed/NCBI
17	Ford A, Al-Magableh M, Gaspari TA and Hart JL: Chronic NaHS treatment is vasoprotective in high-fat-fed ApoE(−/−) mice. Int J Vasc Med. 2013:9159832013.
18	Zhang C, Zheng H, Yu Q, Yang P, Li Y, Cheng F, Fan J and Liu E: A practical method for quantifying atherosclerotic lesions in rabbits. J Comp Pathol. 142:122–128. 2010. View Article : Google Scholar
19	Yu Q, Li Y, Wang Y, Zhao S, Yang P, Chen Y, Fan J and Liu E: C-reactive protein levels are associated with the progression of atherosclerotic lesions in rabbits. Histol Histopathol. 27:529–535. 2012.PubMed/NCBI
20	de Boer RA, Voors AA, Muntendam P, van Gilst WH and van Veldhuisen DJ: Galectin-3: A novel mediator of heart failure development and progression. Eur J Heart Fail. 11:811–817. 2009. View Article : Google Scholar : PubMed/NCBI
21	Liu E, Kitajima S, Higaki Y, Morimoto M, Sun H, Watanabe T, Yamada N and Fan J: High lipoprotein lipase activity increases insulin sensitivity in transgenic rabbits. Metabolism. 54:132–138. 2005. View Article : Google Scholar
22	Meir KS and Leitersdorf E: Atherosclerosis in the apolipo-protein-E-deficient mouse: A decade of progress. Arterioscler Thromb Vasc Biol. 24:1006–1014. 2004. View Article : Google Scholar : PubMed/NCBI
23	Reddick RL, Zhang SH and Maeda N: Atherosclerosis in mice lacking apo E. Evaluation of lesional development and progression. Arterioscler Thromb. 14:141–147. 1994. View Article : Google Scholar : PubMed/NCBI
24	Tupin E, Nicoletti A, Elhage R, Rudling M, Ljunggren HG, Hansson GK and Berne GP: CD1d-dependent activation of NKT cells aggravates atherosclerosis. J Exp Med. 199:417–422. 2004. View Article : Google Scholar : PubMed/NCBI
25	Ni M, Zhang M, Ding SF, Chen WQ and Zhang Y: Micro-ultrasound imaging assessment of carotid plaque characteristics inapolipoprotein-E knockout mice. Atherosclerosis. 197:64–71. 2008. View Article : Google Scholar
26	McAteer MA, Schneider JE, Clarke K, Neubauer S, Channon KM and Choudhury RP: Quantification and 3D reconstruction of atherosclerotic plaque components in apolipoprotein E knockout mice using ex vivo high-resolution MRI. Arterioscler Thromb Vasc Biol. 24:2384–2390. 2004. View Article : Google Scholar : PubMed/NCBI
27	Wrobel TP, Mateuszuk L, Kostogrys RB, Chlopicki S and Baranska M: Quantification of plaque area and characterization of plaque biochemical composition with atherosclerosis progression in ApoE/LDLR(^-/^-) mice by FT-IR imaging. Analyst. 138:6645–6652. 2013. View Article : Google Scholar : PubMed/NCBI