The Effect of Flaxseed (Linum usitatissimum) Ethanol Extract on LDL Cholesterol Levels and Foam Cell Number in The Aorta of Male Wistar Rats (Rattus norvegicus) Fed A High-Fat and High-Fructose Diet

Marzha Marcella, Desak Made Wihandani, I Gusti Ayu Artini, I Wayan Juli Sumadi, I Made Pande Dwipayana, Ni Putu Sriwidyani

Abstract

A diet high in saturated fat and fructose leads to dyslipidemia, increasing atherosclerosis risk. Although lipid-lowering medications are available, they have certain limitations. Flaxseed, rich in ALA, lignants, and phytosterols, may improve lipid profiles and act as an antioxidant. This study evaluated the effectiveness of flaxseed in preventing LDL cholesterol elevation and aortic foam cell formation in male Wistar rats fed a high-fat, high-fructose (HFHF) diet.This true experimental study used a post-test-only control group design for 35 days. Twenty-five male Wistar rats were randomly assigned to five groups: N (normal control, standard diet), K (negative control, HFHF diet), and three treatment groups (P1, P2, P3, HFHF diet) receiving flaxseed ethanol extract at doses of 200, 400, and 800 mg/kgBW/day. LDL cholesterol levels (mg/dL) and aortic foam cell counts (cells/HPF) were measured. Data were analyzed using Shapiro-Wilk and Levene tests for normality and homogeneity, followed by one-way ANOVA, Kruskal-Wallis, and Tukey-HSD post hoc tests. LDL cholesterol levels in groups N, P1, P2, and P3 were significantly lower than in group K (p = 0.007), with P2 showing the most significant inhibition (p = 0.035). Foam cell counts were lower in treatment groups than in group K (p = 0.257), but no significant differences were found among them. Flaxseed (Linum usitatissimum) ethanol extract at 400 mg/kgBW/day was the most effective in preventing an increase in LDL cholesterol. However, the extract at all three doses was ineffective in preventing an increase in aortic foam cells in male Wistar rats (Rattus norvegicus) fed an HFHF diet.

Keywords

Atherosclerosis; Dyslipidemia; Flaxseed; Foam cell; LDL

References

Bruen, R., Fitzsimons, S., & Belton, O. (2017). Atheroprotective effects of conjugated linoleic acid. British Journal of Clinical Pharmacology, 83(1), 46–53. https://doi.org/10.1111/bcp.12948

Bujok, J., Mista, D., Wincewicz, E., Kroliczewska, B., Dzimira, S., & Zuk, M. (2021). Atherosclerosis Development and Aortic Contractility in Hypercholesterolemic Rabbits Supplemented with Two Different Flaxseed Varieties. Bioactive/Nutraceutical Compounds in Plant Foods, 10(3), Article 534. https://doi.org/10.3390/foods10030534

Chahirou, Y., Lamtai, M., Mesfioui, A., Ouichou, A., Coulibaly, M., Boussekkour, R., & El Hessni, A. (2018). Impact of the Association of a High Fructose Diet and Chronic Mild Stress on Metabolic and Affective Disorders in Male Rat. Journal of Behavioral and Brain Science, 08(04), 157–170. https://doi.org/10.4236/jbbs.2018.84010

Chan, J., Karere, G. M., Cox, L. A., & VandeBerg, J. L. (2015). Animal Models of Diet-induced Hypercholesterolemia. InTech. https://doi.org/10.5772/59610

Djuricic, I., & Calder, P. C. (2021). Beneficial Outcomes of Omega-6 and Omega-3 Polyunsaturated Fatty Acids on Human Health: An Update for 2021. Nutrients, 13(7), Article 2421. https://doi.org/10.3390/nu13072421

Es-said, S., Lafhal, K., Elkhiat, A., Hammoud, M., Regbaoui, N., Ezoubeiri, A., Makbal, R., Sbyea, S., Elhiba, O., Sellami, S., Rais, H., Karim, A., Gamrani, H., Rada, N., Bouskraoui, M., & Fdil, N. (2022). Flaxseed extract reduces tissue accumulation and enhances urinary excretion of chondroitin sulphate in the rat: a possible new path in substrate reduction therapy for mucopolysaccharidosis. Pharmaceutical Biology, 60(1), 879–888. https://doi.org/10.1080/13880209.2022.2068618

Fadli, J. C., Guntarti, A., Salamah, N. (2024). Autentikasi Minyak Biji Jagung Zea mays L.: Pengaruh Metode Ekstraksi dan Analisis Komposisi Menggunakan Metode Gas Cromatography-Mass Spectrometry (GC-MS). Journal of Pharmaceutical Science and Clinical Research, 9(2), 256-267. https://doi.org/10.20961/jpscr.v9i2.85367

Farida, Y., Putri, C.I.P. (2016). Efek Penggunaan Simvastatin terhadap Kenaikan Gula Darah Puasa Pasien Diabetes Melitus Tipe 2. Journal of Pharmaceutical Science and Clinical Research, 1(1), 58-65.

Febrianingsih, E. (2019). Pengaruh Pemberian Diet High Fat High Fructose terhadap Jumlah Sel Busa dan Ketebalan Dinding pada Aorta Tikus Sprague Dawley Jantan Model Obesitas Pengaruh Pemberian Diet High Fat High Fructose terhadap Jumlah Sel Busa dan Ketebalan Dinding pada Aorta Tikus Sprague Dawley Jantan Model Obesitas (Thesis). Universitas Brawijaya, Malang.

Gileva, O. G., Butolin, E. G., Tereshchenko, M. V., & Ivanov, V. G. (2022). Evaluation of indicators of carbohydrate and lipid metabolism in rats depending on the type of high-calorie diet. Obesity and Metabolism, 19(1), 47–52. https://doi.org/10.14341/omet12712

Gurina, T. S., & Simms, L. (2025). Histology, Staining. Florida: StatPearls Publishing. https://www.ncbi.nlm.nih.gov/books/NBK557663/

Han, H., Yan, P., Chen, L., Luo, C., Gao, H., Deng, Q., Zheng, M., Shi, Y., & Liu, L. (2015). Flaxseed Oil Containing α -Linolenic Acid Ester of Plant Sterol Improved Atherosclerosis in ApoE Deficient Mice. Oxidative Medicine and Cellular Longevity, 2015, 1–17. https://doi.org/10.1155/2015/958217

Handayani, D., Febrianingsih, E., Desi Kurniawati, A., Kusumastuty, I., Nurmalitasari, S., Widyanto, R. M., Oktaviani, D. N., Maghfirotun Innayah, A., & Sulistyowati, E. (2021). High-Fructose Diet Initially Promotes Increased Aortic Wall Thickness, Liver Steatosis, and Cardiac Histopathology Deterioration, but Does Not Increase Body Fat Index. Journal of Public Health Research, 10(2), Article 2181. https://doi.org/10.4081/jphr.2021.2181

Hardimarta, F. P., Ikawati, K., & Yuniarti, C. A. (2020). Perbaikan Gambaran Lesi Aterosklerotik Dengan Pemberian Ekstrak Beta Vulgaris Pada Tikus Model Diet Atherogenik. Media Farmasi Indonesia, 15(1), 1571-1577. https://doi.org/10.53359/mfi.v15i1.140

Harsa, I. M. S. (2014). Efek Pemberian Diet Tinggi Lemak terhadap Profil Lemak Darah Tikus Putih (Rattus novergicus). Jurnal Ilmiah Kedokteran, 3(1), 21-28.

Indonesian Endocrinology Association. (2021). Pedoman Pengelolaan Dislipidemia di Indonesia. Available from: https://pbperkeni.or.id/wp-content/uploads/2022/02/23-11-21-Website-Panduan-Dislipidemia-2021-Ebook.pdf . [Accessed 5th April 2024]

Indonesian Heart Association. (2022). Panduan Tata Laksana Dislipidemia. Available from: https://www.inaheart.org/storage/guideline/7f75cffa237e402536f3d425012d985d.pdf. [Accessed 5th April 2024]

Institutional Animal Care and Use Program. (2023). Oral Gavage in Mice and Rats IACUC Standard Procedure. Available from: https://iacuc.ucsf.edu/sites/g/files/tkssra16261/files/wysiwyg/STD%20PROCEDURE%20-%20Misc%20Rodent%20Procedures%20-%20Oral%20Gavage%20In%20Mice%20and%20Rats.pdf. [Accessed 10th January 2025]

Juslim, R. R., & Herawati, F. (2018). Penyakit Kardiovaskular. Yogyakarta: Graha Ilmu.

Juszczyk, G., Mikulska, J., Kasperek, K., Pietrzak, D., Mrozek, W., & Herbet, M. (2021). Chronic Stress and Oxidative Stress as Common Factors of the Pathogenesis of Depression and Alzheimer’s Disease: The Role of Antioxidants in Prevention and Treatment. Antioxidants, 10(9), Article 1439. https://doi.org/10.3390/antiox10091439

Kanikowska, D., Korybalska, K., Mickiewicz, A., Rutkowski, R., Kuchta, A., Sato, M., Kreft, E., Fijałkowski, M., Gruchała, M., Jankowski, M., Breborowicz, A., & Witowski, J. (2020). Flaxseed (Linum Usitatissimum L.) Supplementation in Patients Undergoing Lipoprotein Apheresis for Severe Hyperlipidemia—A Pilot Study. Nutrients, 12(4), Article 1137. https://doi.org/10.3390/nu12041137

Kauser, S., Hussain, A., Ashraf, S., Fatima, G., Ambreen, Javaria, S., Abideen, Z. U., Kabir, K., Yaqub, S., Akram, S., Shehzad, A., & Korma, S. A. (2024). Flaxseed (Linum usitatissimum); phytochemistry, pharmacological characteristics and functional food applications. Food Chemistry Advances, Article 100573. https://doi.org/10.1016/j.focha.2023.100573

Kawakami, Y., Yamanaka-Okumura, H., Naniwa-Kuroki, Y., Sakuma, M., Taketani, Y., & Takeda, E. (2015). Flaxseed oil intake reduces serum small dense low-density lipoprotein concentrations in Japanese men: a randomized, double blind, crossover study. Nutrition Journal, 14(1), Article 39. https://doi.org/10.1186/s12937-015-0023-2

Kinanti, A. P., Lestari, A., Nabilah, Z. M., Maulida, R., Widiastuti, T. C., Kiromah, N. Z. W. (2023). Uji Aktivitas Antidiabetes Ekstrak Etanol Daun Ganitri (Elaeocarpus ganitrus Roxb.) Pada Tikus Wistar Jantan (Rattus norvegicus) Yang Diinduksi Streptozotocin. Journal of Pharmaceutical Science and Clinical Research, 8(1), 139-151. https://doi.org/10.20961/jpscr.v8i1.64771

Kumar, N., Aithal, A. P., & Verma, S. (2021). Morphohistological analysis of the prevalence and distribution of atheroma in the abdominal aorta and its branches: a cadaveric study. Journal Vascular Brasileiro, 20, Article e20210014. https://doi.org/10.1590/1677-5449.210014

Kurniawan, D. W., Lestari, N. D., Sulistyo, H., Cacu. (2022). Pengaruh Pemberian Kombinasi Ekstrak Etanol Brotowali, Sambiloto, Meniran dan Kayu Manis Terhadap Histopatologi Glomerulus Tikus Model Hiperglikemia. Journal of Pharmaceutical Science and Clinical Research, 7(3), 294-306. https://doi.org/ 10.20961/jpscr.v7i3.51501

Li, M. Q., Jiao, Q. Z., Xin, W. Q., Niu, S. L., Liu, M. M., Song, Y. X., Wang, Z. G., Yang, X. Y., & Liang, D. G. (2022). The Emerging Role of Rho Guanine Nucleotide Exchange Factors in Cardiovascular Disorders: Insights into Atherosclerosis: A Mini Review. Frontiers Cardiovascular Medicine, 8, Article 782098. https://doi.org/10.3389/fcvm.2021.782098

Maghfiroh, R. M., Hariani, D., & Khaleyla, F. (2022). Efektivitas Ekstrak Daun Pepaya Jepang (Cnidoscolus aconitifolius) terhadap Kadar Kolesterol dan Histologi Aorta Mencit Hiperkolesterolemia. LenteraBio, 11(1), 89–100. https://doi.org/10.26740/lenterabio.v11n1.p89-100

Malik, V. S., & Hu, F. B. (2015). Fructose and Cardiometabolic Health. Journal of the American College of Cardiology, 66(14), 1615–1624. https://doi.org/10.1016/j.jacc.2015.08.025

Ma’rufi, R., & Rosita, L. (2014). Hubungan Dislipidemia dan Kejadian Penyakit Jantung Koroner. JKKI, 6(1), 48–52.

Mercola, J., & D’Adamo, C. R. (2023). Linoleic Acid: A Narrative Review of the Effects of Increased Intake in the Standard American Diet and Associations with Chronic Disease. Nutrients, 15(14), Article 3129. https://doi.org/10.3390/nu15143129

Ministry of Health Republic Indonesia. (2018). Laporan Riset Kesehatan Dasar. Available from: https://repository.badankebijakan.kemkes.go.id/id/eprint/3514/1/Laporan%20Riskesdas%202018%20Nasional.pdf. [Accessed 24th April 2024].

Moss, J. W. E., Davies, T. S., Garaiova, I., Plummer, S. F., Michael, D. R., & Ramji, D. P. (2016). A Unique Combination of Nutritionally Active Ingredients Can Prevent Several Key Processes Associated with Atherosclerosis In Vitro. PLoS ONE, 11, Article e0151057. https://doi.org/10.1371/journal.pone.0151057

Mueed, A., Shibli, S., Korma, S. A., Madjirebaye, P., Esatbeyoglu, T., & Deng, Z. (2022). Flaxseed Bioactive Compounds: Chemical Composition, Functional Properties, Food Applications and Health Benefits-Related Gut Microbes. Foods, 11(20), Article 3307. https://doi.org/10.3390/foods11203307

Pizzini, A., Lunger, L., Demetz, E., Hilbe, R., Weiss, G., Ebenbichler, C., & Tancevski, I. (2017). The Role of Omega-3 Fatty Acids in Reverse Cholesterol Transport: A Review. Nutrients, 9(10), Article 1099. https://doi.org/10.3390/nu9101099

Polat, E. S., Citil, O. B., & Garip, M. (2013). Fatty acid composition of yolk of nine poultry species kept in their natural environment. Animal Science Papers and Reports, 31(4), 363–368.

Poznyak, A. V., Nikiforov, N. G., Starodubova, A. V., Popkova, T. V., & Orekhov, A. N. (2021). Macrophages and Foam Cells: Brief Overview of Their Role, Linkage, and Targeting Potential in Atherosclerosis Macrophages and Foam Cells: Brief Overview of Their Role, Linkage, and Targeting Potential in Atherosclerosis. Biomedicines, 9(9), Article 1221. https://doi.org/10.3390/biomedicines9091221

Putri, D. A. (2018). Pengaruh Pemberian Ekstrak Etanol Biji Rami (Linum usitatissimum L.) dengan Fibroblast Growth Factor (FGF) Dari Telur Ayam Terfertilisasi Terhadap Kadar Glukosa Darah Mencit Putih Jantan Hiperglikemia (Thesis). STFI Yayasan Perintis, Padang.

Rosmala, A. Z., Asmara, I. G. Y., & Widiastuti, I. A. E. (2018). Perbedaan Hasil Pengukuran Kadar Kolesterol LDL antara Metode Direk dan Indirek dengan Menggunakan Rumus Friedewald pada Tikus Putih (Rattus norvegicus). Unram Medical Journal, 7(3), 1-4. https://doi.org/10.29303/jku.v7i3.178

Sanches, S. C. L., Ramalho, L. N. Z., Augusto, M. J., da Silva, D. M., & Ramalho, F. S. (2015). Nonalcoholic Steatohepatitis: A Search for Factual Animal Models. BioMed Research International, 2015, 1–13. https://doi.org/10.1155/2015/574832

Shahidi, S., Mahmoodi, M. S., Komaki, A., & Sadeghian, R. (2022). The comparison of omega-3 and flaxseed oil on serum lipids and lipoproteins in hyperlipidemic male rats. Heliyon, 8(6), Article e09662. https://doi.org/10.1016/j.heliyon.2022.e09662

Susanti, N., Rahmawati, E., & Aprinda, R. K. (2019). Efek Diet Tinggi Fruktosa terhadap Profil Lipid Tikus Rattus Rattus norvegicus Strain Wistar. Journal of Islamic Medicine, 3(2), 26–35.

Veonika, D. A., Wiboworini, B., Muthmainah. (2024). Cholecalciferol Effects on Lipid Profile of Experimental Animals: A Scoping Review. Journal of Pharmaceutical Science and Clinical Research, 9(1), 33-44. https://doi.org/10.20961/jpscr.v9i1.73400

Wahyuni, S. (2015). Dislipidemia (1st ed., Vol. 1). Denpasar: Udayana University Press.

World Health Organization. (2021). Cardiovascular diseases (CVDs). Available from: https://www.who.int/news-room/fact-sheets/detail/cardiovascular-diseases-(cvds). [Accessed 24th July 2024].

Xu, L. Z. , Gao, F., Yang, Z. L., Han, L.J., & Liu, X. (2016). Discriminant analysis of terrestrial animal fat and oil adulteration in fish oil by infrared spectroscopy. Int J Agric & Biol Eng, 9(3), 179-185. https://doi.org/10.3965/j.ijabe.20160903.2239.

Yang, C., Xia, H., Wan, M., Lu, Y. F., Xu, D. F., Yang, X., Yang, L. G., & Sun, G. J. (2021). Comparisons of the effects of different flaxseed products consumption on lipid profiles, inflammatory cytokines and anthropometric indices in patients with dyslipidemia related diseases: systematic review and a dose–response meta-analysis of randomized controlled trials. Nutrition & Metabolism, 18(1), Article 91. https://doi.org/10.1186/s12986-021-00619-3

DOI: https://doi.org/10.20961/jpscr.v10i3.100142

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