Prolonged-heated High-Fat Diet Increase the Serum LDL Cholesterol Level and Induce the Early Atherosclerotic Plaque Development in Wistar Rats


  • Valentina Yurina
  • Ema Pristi Yunita
  • Tri Yudani Mardining Raras
  • Achmad Rudijanto
  • Kusworini Handono


Cholesterol, foam cells, high-fat diet (HFD), oxidized lipid, atherosclerosis


Rats are one of the most widely used animal models in health research. However, since rats are relatively resistant to atherosclerosis, the transgenic rats often used to study atherosclerosis in rats. Our study suggests a prolonged-heated lipid to induce atherosclerosis in rats, therefore provide a more low-cost option to study atherosclerosis in rats. Aims of this study is to compare the effect of heated high-fat diet (HFD) to the LDL and HDL cholesterol level and foam cell formation in the Wistar rat animal model. Rats were divided into three groups, control group was given with the normal diet, and the other two treated groups received oxidized HFD (heated HFD) and HFD, respectively. The heated HFD contain lard that was previously heated at 190°C for 24 h. Diet was given for 8 weeks. The serum LDL and HDL cholesterol level were measured before and 4th and 8th weeks after treatment with help of colorimetric measurement. The aorta analyzed for the foam cell formation after HE staining using the light microscope.  The prolonged- heated HFD significantly increased the LDL cholesterol from the 4th week of the treatment (p = 0.023). The rats aortic from HFD and heated HFD treated groups showed a mononuclear cells infiltration and the early foam cell formation. The heating process of the lipid caused lipid oxidation which significantly increased the LDL cholesterol level of the animal model and foam cells formation. This study suggests that Wistar rats with heated HFD could demonstrated early atherosclerosis plaque development.


van der Ende MY, Hartman MHT, Hagemeijer Y et al. (2017) The lifelines cohort study: Prevalence and treatment of cardiovascular disease and risk factors. International Journal of Cardiology 228 495 – 500. doi: 10.1016/j.ijcard.2016.11.061.

Lloyd-Jones DM, Leip EP, Larson MG et al. (2006) Prediction of lifetime risk for cardiovascular disease by risk factor burden at 50 years of age. Circulation 113: 791–798. doi: 10.1161/CIRCULATIONAHA.105.548206.

Libby P (2003) Vascular biology of atherosclerosis: Overview and state of the art. The American journal of cardiology 91 (2): 3A – 6A. doi: 10.1016/S0002-9149(02)03143-0.

Frostegård J (2013) Immunity, atherosclerosis and cardiovascular disease. BMC Medicine 11: 117. doi: 10.1186/1741-7015-11-117.

Vaziri ND (2014) Role of dyslipidemia in impairment of energy metabolism, oxidative stress, inflammation and cardiovascular disease in chronic kidney disease. Clinical and Experimental Nephrology 18 (2): 265 – 268. doi: 10.1007/s10157-013-0847-z.

Gami AS, Witt BJ, Howard DE et al. (2007) Metabolic syndrome and risk of incident cardiovascular events and death: A systematic review and meta-analysis of longitudinal studies. Journal of the American College of Cardiology 49 (4): 403–414. doi: 10.1016/j.jacc.2006.09.032.

Bonomini F, Tengattini S, Fabiano A et al. (2008) Atherosclerosis and oxidative stress. Histology and Histopathology 23 (3): 381–390. doi: 10.14670/HH-23.381.

Moriwaki H, Kume N, Sawamura T et al. (1998) Ligand specificity of LOX-1, a novel endothelial receptor for oxidized low density lipoprotein. Arteriosclerosis, Thrombosis, and Vascular Biology 18 (10): 1541 – 1547. doi: 10.1161/01.ATV.18.10.15

Sugiyama D, Higashiyama A, Wakabayashi I et al. (2015) The relationship between lectin-like oxidized low-density lipoprotein receptor-1 ligands containing apolipoprotein B and the cardio-ankle vascular index in healthy commu-nity inhabitants: The KOBE study. Journal of Atheroscle-rosis and Thrombosis 22 (5): 499 – 508. doi: 10.5551/jat.26450.

Mango R, Predazzi IM, Romeo F, Novelli G (2011) LOX-1/LOXIN: The yin/yang of atheroscleorosis. Cardiovascular drugs and therapy / sponsored by the International Society of Cardiovascular Pharmacotherapy 25 (5): 489 – 494. doi: 10.1007/s10557-011-6333-5.

Rader DJ, Puré E (2005) Lipoproteins, macrophage function, and atherosclerosis: Beyond the foam cell? Cell Metabolism 1 (4): 223 – 230. doi: 10.1016/j.cmet.2005.03.005.

Eder K, Keller U, Hirche F, Brandsch C (2003) Thermally oxidized dietary fats increase the susceptibility of rat LDL to lipid peroxidation but not their uptake by macrophages. The Journal of Nutrition 133 (9): 2830 – 2837. doi: 10.1093/jn/133.9.2830.

Duelsner A, Persson AB (2013) Animal models in cardiovascular research. Acta Physiologica 208 (1): 1 – 5. doi: 10.1111/


Kapourchali FR (2014) Animal models of atherosclerosis. World Journal of Clinical Cases 2 (5): 126 – 132. doi: 10.12998/wjcc.v2.i5.126.

Xiangdong L, Yuanwu L, Hua Z et al. (2011) Animal models for the atherosclerosis research: A review. Protein and Cell 2 (3): 189 – 201. doi: 10.1007/s13238-011-1016-3.

Nawar WW (1984) Chemical changes in lipids produced by thermal processing. Journal of Chemical Education 61 (4): 299 – 302. doi: 10.1021/ed061p299.

Riahi S, Mohammadi MT, Sobhani V et al. (2015) Chronic effects of aerobic exercise on gene expression of LOXâ€1 receptor in the heart of rats fed with high fat diet. Iranian Journal of Basic Medical Sciences 18 (8): 805 – 812.

Fruchart JC, Nierman MC, Stroes ESG et al. (2004) New risk

factors for atherosclerosis and patient risk assessment. Circulation 109: III15 – I19. doi: 10.1161/01.CIR.0000131513.33892.5b.

Venkata RM, Subramanyam R (2016) Evaluation of the deleterious health effects of consumption of repeatedly heated vegetable oil. Toxicology Reports 3: 636 – 643. doi: 10.1016/j.toxrep.2016.08.003.

Escolà -Gil JC, Llaverias G, Julve J et al. (2011) The cholesterol content of western diets plays a major role in the paradoxical increase in high-density lipoprotein cholesterol and upregulates the macrophage reverse cholesterol transport pathway. Arteriosclerosis, Thrombosis, and Vascular Biology 31 (11): 2493–2499. doi: 10.1161/ATVBAHA.111.236075.

Tréguier M, Briand F, Boubacar A et al. (2011) Diet-induced dyslipidemia impairs reverse cholesterol transport in hamsters. European Journal of Clinical Investigation 41 (9): 921 – 928. doi: 10.1111/j.1365-2362.2011.02478.x.

Libby P, Ridker PM, Hansson GK (2011) Progress and challenges in translating the biology of atherosclerosis. Nature 473: 317 – 325. doi: 10.1038/nature10146.

Khan-Merchant N, Penumetcha M, Meilhac O, Parthasarathy S (2002) Oxidized fatty acids promote atherosclerosis only in the presence of dietary cholesterol in low-density lipoprotein receptor knockout mice. The Journal of Nutrition 132 (11): 3256 – 3262. doi: 10.1093/jn/132.11.3256

Libby P, Ridker PM, Hansson GK (2009) Inflammation in atherosclerosis from pathophysiology to practice. Journal of the American College of Cardiology 54 (23): 2129 – 2138. doi: 10.1016/j.jacc.2009.09.009.

Weber C, Noels H (2011) Atherosclerosis: current pathogenesis and therapeutic options. Nature Medicine 17 (11): 1410 – 1422. doi: 10.1038/nm.2538.

Clarke M, Bennett M (2006) The emerging role of vascular smooth muscle cell apoptosis in atherosclerosis and plaque stability. American Journal of Nephrology 26 (6): 531–535. doi: 10.1159/000097815.

Ishino S, Mukai T, Kume N et al. (2007) Lectin-like oxidized LDL receptor-1 (LOX-1) expression is associated with atherosclerotic plaque instability — analysis in hypercholesterolemic rabbits. Atherosclerosis 195 (1): 48 – 56. doi: 10.1016/j.atherosclerosis.2006.11.031.

Rudijanto A (2007) The role of vascular smooth muscle cells on the pathogenesis of atherosclerosis. Acta Medica Indonesiana 39 (2): 86 – 93.

Kucera O, Cervinkova Z (2014) Experimental models of non-alcoholic fatty liver disease in rats. World Journal of Gastroenterology 20 (26): 8364 – 8376. doi: 10.3748/wjg.v20.i26.8364.

Van Herck MA, Vonghia L, Francque SM (2017) Animal models of nonalcoholic fatty liver disease—a starter’s guide. Nutrients 9 (10): 1 – 13. doi: 10.3390/nu9101072.