Effect of Fish Oil on Liver MMP-9 and MMP-12 Levels in Hypercholesterolemia Induced Mice

Abstract

Aim: This study aimed to investigate the effects of fish oil on liver matrix metalloproteinase-9 (MMP-9) and MMP-12 levels in hypercholesterolemic mice. Materials and Methods: For comprehensive evaluation of MMP-9 and MMP-12 levels, total cholesterol (TC), low-density lipoprotein (LDL), total antioxidan status (TAS) and total oxidan status (TOS) levels, as well as liver histopathological status, were assessed. Twenty-eight mice were equally divided into four groups. The control (CNT) group received standard chow and drinking water without any treatment. The fish oil (FO) group received orally 0.1 ml of fish oil daily for 10 weeks. The cholesterol (CHOL) group was fed with a high-cholesterol chow diet. The cholesterol plus fish oil (CHOL+FO) group received a high-cholesterol chow plus an oral FO solution. Results: The MMP-9, TC and TOS levels of the CHOL group were significantly higher, while TAS were lower compared to other groups. While MMP-9 levels decreased in the CHOL+FO and FO groups compared to the CHOL group (p<0.001), no significant change was observed for MMP-12 (p>0.05). Total oxidan status levels decreased, and TAS levels increased in the CHOL+FO and FO groups compared with the CHOL group, with higher oxidative stress index (OSI) levels (p<0.05). Conclusion: It was concluded that combined fish oil administration could reduce the adverse effects of hypercholesterolemia by down-regulating bad cholesterol and oxidative stress in mouse liver and colon tissue, and by reducing MMP-9 protein concentration in colon tissue.

Keywords

• hypercholesterolemia
• MMP-9
• MMP-12
• fish oil; mouse

References

1. 1. Luo J, Yang H, Song BL. Mechanisms and regulation of cholesterol homeostasis. Nature reviews Molecular Cell Biology. 2020; 21(4):225-245.
2. 2. Bai, X., Yao, M., Zhu, X., Lian, Y., Zhang, M. (2023). Baicalin suppresses interleukin-1β-induced apoptosis, inflammatory response, oxidative stress, and extracellular matrix degradation in human nucleus pulposus cells. Immunopharmacology and Immunotoxicology 45(4), 433-442.
3. 3. Wang K, Zhang Y, Si C, Cao Y, Shao P, Zhang P, et al. Cholesterol: The driving force behind the remodeling of tumor microenvironment in colorectal cancer. Heliyon. 2024;10(23).
4. 4. Khotimchenko YS, Silachev DN, Katanaev VL. Marine natural products from the Russian Pacific as sources of drugs for neurodegenerative diseases. Marine Drugs. 2022;20(11):708.
5. 5. Nazir M, Al-Ansari A, Al-Khalifa K, Alhareky M, Gaffar B, Almas K. Global prevalence of periodontal disease and lack of its surveillance. The Scientific World Journal. 2020;2020(1):2146160.
6. 6. Chuliá-Peris L, Carreres-Rey C, Gabasa M, Alcaraz J, Carretero J, Pereda J. Matrix metalloproteinases and their inhibitors in pulmonary fibrosis: EMMPRIN/CD147 comes into play. International Journal of Molecular Sciences 2022; 23(13):6894.
7. 7. Karapehlivan M, Ogun M, Kaya I, Ozen H, Deveci HA, Karaman M. Protective effect of omega-3 fatty acid against mercury chloride intoxication in mice. Journal of Trace Elements in Medicine and Biology 2014;28(1):94-99.
8. 8. Chen J, Jayachandran M, Bai W, Xu B. A critical review on the health benefits of fish consumption and its bioactive constituents. Food Chemistry 2022;369:130874.

9. 9. Kaya I, Citil M, Sozmen M, Karapehlivan M, Cigsar G. Investigation of protective effect of L-carnitine on L-asparaginase-induced acute pancreatic injury in male Balb/c mice. Digestive Diseases and Sciences 2015;60:1290-1296.
10. 10. Çenesiz S. The role of oxidant and antioxidant parameters in the infectious diseases: A systematic literature review. Kafkas Universitesi Veteriner Fakultesi Dergisi. 2020;26(6).
11. 11. Kaya, MM. The roles of matrix metalloproteases in different stages of cancer. In: Kaya I. Current Researches in Cancer Biology. Livre de Lion. 2023;27.
12. 12. Erel O. A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry 2004; 37:277-285.
13. 13. Erel O. A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry 2005;38:1103-11.
14. 14. Deveci HA, Unal S, Karapehlivan M, Ayata MK, Gaffaroglu M, Kaya I, Yilmaz M. Effects of glyphosate (herbicide) on serum paraoxonase activity, high density lipoprotein, total antioxidant and oxidant levels in Kars Creek Transcaucasian Barbs (Capoeta capoeta [Guldenstaedt, 1773]) Fresenius Environmental Bulletin (2017); 3514-3518.
15. 15. Chacon AC, Phillips BE, Chacon MA, Brunke-Reese D, Kelleher SL, et al. Oral omega-3 fatty acids promote resolution in chemical peritonitis. Journal of Surgical Research 2016; 206(1):190-198.
16. 16. Penumathsa SV, Thirunavukkarasu M, Koneru S, Juhasz B, Zhan L, Pant R, et al. Statin and resveratrol in combination induces cardioprotection against myocardial infarction in hypercholesterolemic rat. Journal of Molecular and Cellular Cardiology. 2007;42(3):508-516, 2007.
17. 17. Du Y, Su J, Yan M, Wang Q, Wang T, Gao S, et al. Polymethoxyflavones in citrus extract has a beneficial effect on hypercholesterolemia rats by promoting liver cholesterol metabolism. Journal of Ethnopharmacology. 2024;322:117644.
18. 18. Hofker MH, van Vlijmen BJ, Havekes LM. Transgenic mouse models to study the role of APOE in hyperlipidemia and atherosclerosis. Atherosclerosis1998;137(1):1-11.
19. 19. Al‐Mansoub MA, Asmawi MZ, Murugaiyah V. Effect of extraction solvents and plant parts used on the antihyperlipidemic and antioxidant effects of Garcinia atroviridis: a comparative study. Journal of the Science of Food and Agriculture 2014;94(8):1552-1558.
20. 20. Mjaatveit M, Oldernes H, Gudbrandsen OA. Effects of diets containing fish oils or fish oil concentrates with high cetoleic acid content on the circulating cholesterol concentration in rodents. A systematic review and meta-analysis. British Journal of Nutrition. 2024;131(4):606-621.
21. 21. Mildner‐Szkudlarz S, Bajerska J. Protective effect of grape byproduct‐fortified breads against cholesterol/cholic acid dietinduced hypercholesterolaemia in rats. Journal of the Science of Food and Agricultur. 2013; 93(13):3271-3278.
22. 22. Leopold JA, Loscalzo J. Oxidative risk for atherothrombotic cardiovascular disease. Free Radical Biology and Medicine. 2009; 47(12):1673-1706. 23. Tall AR, Yvan-Charvet L. Cholesterol, inflammation and innate immunity. Nature Reviews Immunology 2015; 15(2):104-116.
23. 24. Alkan I, Bakırhan EG, Tüfek NH, Arslan G, Altunkaynak BZ, Kaplan S. Investigation of the protective effect of thymoquinone on the livers of the obese rats ınduced by a high-fat diet. Kafkas Journal of Medical Sciences 2024;14(3):252-259.
24. 25. Kamisako T, Tanaka Y, Ikeda T, Yamamoto K, Ogawa H. Dietary fish oil regulates gene expression of cholesterol and bile acid transporters in mice. Hepatology Research. 2012; 42(3):321-326.
25. 26. Nishimoto T, Pellizzon MA, Aihara M, Stylianou IM, Billheimer JT, Rothblat G, et al. Fish oil promotes macrophage reverse cholesterol transport in mice. Arteriosclerosis, Thrombosis, and Vascular Biology. 2009;29(10):1502-1508.
26. 27. Hirako S, ju Kim H, Arai T, Chiba H, Matsumoto A. Effect of concomitantly used fish oil and cholesterol on lipid metabolism. The Journal of Nutritional Biochemistry 2010;21(7):573-579.
27. 28. Sales ALC, Primo MG, Mello Neto RS, Mendes AVS, Furtado MM, Rocha JÉL, et al. A Preliminary study on hepatoprotective, hypolipidemic and aortic morphometric effects of omega-3rich fish oil in hypercholesterolemic mice. Pharmaceuticals. 2024;17(1):72.
28. 29. Lee YT, Lin HY, Chan YWF, Li KHC, To OTL, Yan BP, et al. Mouse models of atherosclerosis: a historical perspective and recent advances. Lipids in health and disease. 2017;16:1-11.
29. 30. Rader DJ, Tall AR. Is it time to revise the HDL cholesterol hypothesis?. Nature medicine. 2012; 18(9):1344-1346.
30. 31. Nestel PJ. Fish oil and cardiovascular disease: lipids and arterial function. The American Journal of Clinical .000; 71(1):228S-231S.
31. 32. Oelrich B, Dewell A, Gardner CD. Effect of fish oil supplementation on serum triglycerides, LDL cholesterol and LDL subfractions in hypertriglyceridemic adults. Nutrition, metabolism and cardiovascular diseases. 2013;23(4):350-357.
32. 33. Komprda T, Škultéty O, Křížková S, Zorníková G, Rozíková V, Krobot R. Effect of dietary Schizochytrium microalga oil and f ish oil on plasma cholesterol level in rats. Journal of animal physiology and animal nutrition. 2015;99(2):308-316.
33. 34. Shang T, Liu L, Zhou J, Zhang M, Hu Q, Fang M, et al. Protective effects of various ratios of DHA/EPA supplementation on highfat diet-induced liver damage in mice. Lipids in Health and Disease. 2017;16:1-13.
34. 35. Kaya İ, Dağ S, Mavioğlu Kaya, M, Tanrıverdi EA, Beşeren H, Aşasın G. Modulatory effect of pomegranate extract on TRPA1, TRPM2 and caspase-3 expressions in colorectal cancer induction of mice. Turkish Journal of Biochemistry 2022;47(5):612-619.
35. 36. Wang Q, Ji C, Smith P, McCulloch CA. Impact of TRP channels on extracellular matrix remodeling: focus on TRPV4 and collagen. International Journal of Molecular Sciences 2024;25(7):3566.
36. 37. Balkrishna A, Paliwal R, Singh S, Singh R, Gohel V, Dev R, et al. Livogrit mitigates ANIT-induced cholestasis-like symptoms in an in vivo model by curbing hepatic inflammation and regulating BAX, TGF-β, MMP-9 and α-SMA gene expression. Heliyon 2025;11(3).
37. 38. Kalela A, Koivu TA, Höyhtyä M, Jaakkola O, Lehtimäki T, Sillanaukee P, et al. Association of serum MMP-9 with autoantibodies against oxidized LDL. Atherosclerosis 2002; 160(1):161-165.
38. 39. Minami Y, Kawabata K, Kubo Y, Arase S, Hirasaka K, Nikawa, et al. Peroxidized cholesterol-induced matrix metalloproteinase-9 activation and its suppression by dietary β-carotene in photoaging of hairless mouse skin. The Journal of Nutritional Biochemistry. 2009;20(5):389-398.
39. 40. Ferretti G, Bacchetti T, Banach M, Simental-Mendia LE, Sahebkar A. Impact of statin therapy on plasma MMP-3, MMP9, and TIMP-1 concentrations: a systematic review and metaanalysis of randomized placebo-controlled trials. Angiology 2017;68(10):850-862.
40. 41. Hernandez‐Anzaldo S, Brglez V, Hemmeryckx B, Leung D, Filep JG, Vance JE, et al. Novel role for matrix metalloproteinase 9 in modulation of cholesterol metabolism. Journal of the American Heart Association 2016;5(10), e004228.
41. 42. Sabir U, Gu HM, Zhang DW. Extracellular matrix turnover: Phytochemicals target and modulate the dual role of matrix metalloproteinases (MMPs) in liver fibrosis. Phytotherapy Research. 2023;37(11):4932-4962.
42. 43. Johnson JL, George SJ, Newby AC, Jackson CL. Divergent effects of matrix metalloproteinases 3, 7, 9, and 12 on atherosclerotic plaque stability in mouse brachiocephalic arteries. Proceedings of the National Academy of Sciences 2005;102(43):15575-15580.
43. 44. Malaguarnera M, Cauli O, Cabrera-Pastor A. Obesity and adipose-derived extracellular vesicles: implications for metabolic regulation and disease. Biomolecules 2025;15(2):231.
44. 45. Bauters D, Van Hul M, Lijnen HR. Macrophage elastase (MMP-12) in expanding murine adipose tissue. Biochimica et Biophysica Acta (BBA)-General Subjects 2013;1830(4):2954-2959.
45. 46. Song, M., Zhang, S., Tao, Z., Li, J., Shi, Y., Xiong, Y., ... & Chen, S. (2021). MMP-12 siRNA improves the homeostasis of the small intestine and metabolic dysfunction in high-fat diet feeding-induced obese mice. Biomaterials 278, 121183.
46. 47. Arikan MC, Shapiro SD, Mariani TJ. Induction of macrophage elastase (MMP‐12) gene expression by statins. Journal of Cellular Physiology. 2005;204(1):139-145.
47. 48. Yamada S, Wang KY, Tanimoto A, Fan J, Shimajiri S, Kitajima S, et al. Matrix metalloproteinase 12 accelerates the initiation of atherosclerosis and stimulates the progression of fatty streaks to f ibrous plaques in transgenic rabbits. The American Journal of Pathology. 2008; 172(5):1419-1429.
48. 49. Prasad K, Mishra M. Mechanism of hypercholesterolemiainduced atherosclerosis. Reviews in Cardiovascular Medicine. 2022;23(6):212.
49. 50. Kara AB. Pioglitazone Reduces Oxidative Stress in Kidney Against High Sucrose Diet-Induced Metabolic Syndrome in Rats. Kafkas Journal of Medical Sciences 2018;8(3):165-171.
50. 51. Prasad K. Reduction of serum cholesterol and hypercholesterolemic atherosclerosis in rabbits by secoisolariciresinol diglucoside isolated from flaxseed. Circulation. 1999;99(10):1355-1362.
51. 52. Karaca T, Bayıroğlu F, Cemek M, Comba B, Ayaz A, Karaboğa İ. Effects of green tea extract and lactobacillus casei strain shirota on levels of serum minerals, cholesterol, triglycerides, glucose and lactate in rats fed on high-carbohydrate and highlipid diets. Kafkas Journal of Medical Sciences 2013;(1): 1-7.
52. 53. Kaya I, Deveci H, Ekinci U, Karapehlivan M, Kaya M, Alpay M. The Effect of ellagic acid and sodium fluoride ıntake on total sialic acid levels and total oxidant/antioxidant status in mouse testicular tissue. Annual Research & Review in Biology 2015;7(5):329-335.
53. 54. Duarte MM, Rocha JB, Moresco RN, Duarte T, Da Cruz IB, Loro VL, et al Schetinger, M. R. Association between ischemiamodified albumin, lipids and inflammation biomarkers in patients with hypercholesterolemia. Clinical Biochemistry 2009;42(7-8):666-671.
54. 55. Uetake Y, Ikeda H, Irie R, Tejima K, Matsui H, Ogura S, et al. High-salt in addition to high-fat diet may enhance inflammation and fibrosis in liver steatosis induced by oxidative stress and dyslipidemia in mice. Lipids in Health and Disease 2015;14(1):6.
55. 56. Moghadamnia D, Mokthari M, Khatamsaz S. Comparison of protective effects of omega3 fish oil and aqueous extract of glycyrrhiza glabra root on thioacetamide ınduced lipid toxicity in male rats. Biomedical and Pharmacology Journal 2016; 9(1):111-119.
56. 57. Erdoğan H, Fadıllıoglu E, Özgöçmen S, Söğüt S, Özyurt B, Akyol O, et al. Effect of fish oil supplementation on plasma oxidant/antioxidant status in rats. Prostaglandins, Leukotrienes and Essential Fatty Acids 2004;71(3):149-152.
57. 58. Sarıyer ET, Baş M, Çolak H, Özkan Yenal N, Unay Demirel Ö, Yüksel M. Comparison of dietary supplementation with krill oil, fish oil, and astaxanthin on an experimental ethanolınduced gastric ulcer model: a biochemical and histological study. Nutrients. 2024; 16(20):3426.