ESCULETIN EFFECTS ON MICE-IMMORTALIZED MESANGIAL CELLS SUBMITTED TO HIGH-GLUCOSE MEDIA
DOI:
https://doi.org/10.56238/arev8n4-078Keywords:
Esculetin, Mesangial Cells, Diabetes Mellitus, Oxidative Stress, Nitrosative StressAbstract
Introduction: Diabetes mellitus (DM) has been a major research focus due to its high prevalence and mortality in Brazil and worldwide. Characterized by hyperglycemia, DM leads to comorbidities like atherosclerosis, retinopathy, neuropathy, and nephropathy. Its pathophysiology involves disrupted enzyme and receptor functions, inflammation, and increased reactive oxygen species (ROS). Esculetin, a coumarin-derived polyphenol, has shown potent antioxidant activity, improving lipid profiles and reducing proinflammatory cytokine synthesis.
Objective: To study the effect of esculetin on immortalized mouse mesangial cells subjected to high-glucose medium. Methods: Immortalized mesangial cells from mice (MiMCs) were cultured in culture plates with DMEM/F12 supplemented with 5% fetal bovine serum. The cells were subsequently allocated into four groups: NG (control, D-glucose 6.7 mM)); NG+ESCt (control treated with esculetin hydrate 10, 25, 50, 100, or 200 μg/mL); HG (high glucose, D-glucose 30 mM); and HG+ESCt (high glucose treated with esculetin hydrate 10, 25, 50, 100, or 200 μg/mL) for 24, 48, or 72 hours. In these groups, we evaluated the viability, cell proliferation, and antioxidant capacity of esculetin in addition to its effects on oxidative and nitrosative stress. Statistical analyses were performed with GraphPad Prism 9.0 software. The results are presented as the means ± SEs, with P < 0.05 indicating statistical significance.
Results: The DPPH test revealed the antioxidant effect of esculetin. ROS production was greater in the HG group than in the NG group, and TBARS production was lower in the treated groups than in the control groups. There was an increase in the amount of catalase enzyme in the HG group compared with the NG group, a decrease in the treated groups compared with the control groups, and increased activity of SOD and GPx enzymes in the ESCt groups.
Conclusion: Our data suggest that esculetin reduces oxidative stress in this experimental model. Future studies may be promising in identifying esculetin as an effective intervention for the prevention and treatment of DM.
Downloads
References
Internation Diabetes Federation. IDF Diabetes Atlas. 10 ed. Brussels, Belgium. 2021.
World Health Organization. Diabetes 2023 [Available from: https://www.who.int/news-room/fact-sheets/detail/diabetes.
Boyle J. Lehninger principles of biochemistry (4th ed.): Nelson, D., and Cox, M. Biochemistry and Molecular Biology Education. 2005;33(1):74-5.
American Diabetes A. Diagnosis and classification of diabetes mellitus. Diabetes care. 2012;35 Suppl 1(Suppl 1):S64-71.
World Health Organization. Global Report On Diabetes. 2016:86.
El-Remessy AB, Abou-Mohamed G, Caldwell RW, Caldwell RB. High glucose-induced tyrosine nitration in endothelial cells: role of eNOS uncoupling and aldose reductase activation. Investigative ophthalmology & visual science. 2003;44(7):3135-43.
Poornima IG, Parikh P, Shannon RP. Diabetic cardiomyopathy: the search for a unifying hypothesis. Circulation research. 2006;98(5):596-605.
Brownlee M. Biochemistry and molecular cell biology of diabetic complications. Nature. 2001;414(6865):813-20.
Oleniuc M, Secara I, Onofriescu M, Hogas S, Voroneanu L, Siriopol D, et al. Consequences of Advanced Glycation End Products Accumulation in Chronic Kidney Disease and Clinical Usefulness of Their Assessment Using a Noninvasive Technique - Skin Autofluorescence. Maedica. 2011;6(4):298-307.
Giacco F, Brownlee M. Oxidative stress and diabetic complications. Circulation research. 2010;107(9):1058-70.
Vallon V, Komers R. Pathophysiology of the diabetic kidney. Compr Physiol. 2011;1(3):1175-232.
Batlle D. Clinical and cellular markers of diabetic nephropathy. Kidney international. 2003;63(6):2319-30.
Fioretto P, Steffes MW, Barbosa J, Rich SS, Miller ME, Mauer M. Is diabetic nephropathy inherited? Studies of glomerular structure in type 1 diabetic sibling pairs. Diabetes. 1999;48(4):865-9.
Griesmacher A, Kindhauser M, Andert SE, Schreiner W, Toma C, Knoebl P, et al. Enhanced serum levels of thiobarbituric-acid-reactive substances in diabetes mellitus. Am J Med. 1995;98(5):469-75.
Kaspar JW, Niture SK, Jaiswal AK. Nrf2:INrf2 (Keap1) signaling in oxidative stress. Free radical biology & medicine. 2009;47(9):1304-9.
Garg SS, Gupta J, Sahu D, Liu CJ. Pharmacological and Therapeutic Applications of Esculetin. Int J Mol Sci. 2022;23(20).
Wishart DS, Feunang YD, Guo AC, Lo EJ, Marcu A, Grant JR, et al. DrugBank 5.0: a major update to the DrugBank database for 2018. Nucleic acids research. 2018;46(D1):D1074-D82.
Landete JM. Dietary intake of natural antioxidants: vitamins and polyphenols. Critical reviews in food science and nutrition. 2013;53(7):706-21.
Li YY, Song YY, Liu CH, Huang XT, Zheng X, Li N, et al. Simultaneous determination of esculin and its metabolite esculetin in rat plasma by LC‒ESI‒MS/MS and its application in pharmacokinetic study. Journal of chromatography B, Analytical technologies in the biomedical and life sciences. 2012;907:27-33.
Lin WL, Wang CJ, Tsai YY, Liu CL, Hwang JM, Tseng TH. Inhibitory effect of esculetin on oxidative damage induced by t-butyl hydroperoxide in rat liver. Archives of toxicology. 2000;74(8):467-72.
Prabakaran D, Ashokkumar N. Protective effect of esculetin on hyperglycemia-mediated oxidative damage in the hepatic and renal tissues of experimental diabetic rats. Biochimie. 2013;95(2):366-73.
Kang KS, Lee W, Jung Y, Lee JH, Lee S, Eom DW, et al. Protective effect of esculin on streptozotocin-induced diabetic renal damage in mice. Journal of agricultural and food chemistry. 2014;62(9):2069-76.
Wang YH, Liu YH, He GR, Lv Y, Du GH. Esculin improves dyslipidemia, inflammation and renal damage in streptozotocin-induced diabetic rats. BMC complementary and alternative medicine. 2015;15:402.
Tianzhu Z, Shumin W. Esculin Inhibits the Inflammation of LPS-Induced Acute Lung Injury in Mice Via Regulation of TLR/NF-kappaB Pathways. Inflammation. 2015;38(4):1529-36.
Aguilar Diaz De Leon J, Borges CR. Evaluation of Oxidative Stress in Biological Samples Using the Thiobarbituric Acid Reactive Substances Assay. Journal of visualized experiments : JoVE. 2020(159).
Ortega-Villasante C, Burén S, Blázquez-Castro A, Barón-Sola Á, Hernández LE. Fluorescent in vivo imaging of reactive oxygen species and redox potential in plants. Free Radical Biology and Medicine. 2018;122:202-20.
Damle VG, Wu K, Arouri DJ, Schirhagl R. Detecting free radicals post viral infections. Free Radical Biology and Medicine. 2022;191:8-23.
Hampl V. Determination of Nitric Oxide by the. Methods in nitric oxide research. 1996;309.
Thomas S, Karalliedde J. Diabetic nephropathy. Medicine. 2019;47(2):86-91.
Sharma D, Bhattacharya P, Kalia K, Tiwari V. Diabetic nephropathy: New insights into established therapeutic paradigms and novel molecular targets. Diabetes research and clinical practice. 2017;128:91-108.
Ojo OA, Ibrahim HS, Rotimi DE, Ogunlakin AD, Ojo AB. Diabetes mellitus: From molecular mechanism to pathophysiology and pharmacology. Medicine in Novel Technology and Devices. 2023;19:100247.
Godin DV, Wohaieb SA, Garnett ME, Goumeniouk AD. Antioxidant enzyme alterations in experimental and clinical diabetes. Molecular and Cellular Biochemistry. 1988;84(2):223-31.
Wei W, Liu Q, Tan Y, Liu L, Li X, Cai L. Oxidative stress, diabetes, and diabetic complications. Hemoglobin. 2009;33(5):370-7.
Bert P. La Pression Barométrique: Recherches de Physiologie Expérimentale. Paris: G. Masson; 1878.
Harman D. Aging: a theory based on free radical and radiation chemistry. J Gerontol. 1956;11(3):298-300.
McCord JM, Fridovich I. Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem. 1969;244(22):6049-55.
Phillips M, Cataneo RN, Cheema T, Greenberg J. Increased breath biomarkers of oxidative stress in diabetes mellitus. Clin Chim Acta. 2004;344(1-2):189-94.
Rodrigues AM, Bergamaschi CT, Araujo RC, Mouro MG, Rosa TS, Higa EM. Effects of training and nitric oxide on diabetic nephropathy progression in type I diabetic rats. Experimental biology and medicine. 2011;236(10):1180-7.
Medved I, Brown MJ, Bjorksten AR, Murphy KT, Petersen AC, Sostaric S, et al. N-acetylcysteine enhances muscle cysteine and glutathione availability and attenuates fatigue during prolonged exercise in endurance-trained individuals. J Appl Physiol (1985). 2004;97(4):1477-85.
Nogueira GB, Punaro GR, Oliveira CS, Maciel FR, Fernandes TO, Lima DY, et al. N-acetylcysteine protects against diabetic nephropathy through control of oxidative and nitrosative stress by recovery of nitric oxide in rats. Nitric Oxide. 2018;78:22-31.
Punaro GR, Maciel FR, Rodrigues AM, Rogero MM, Bogsan CS, Oliveira MN, et al. Kefir administration reduced progression of renal injury in STZ-diabetic rats by lowering oxidative stress. Nitric oxide : biology and chemistry. 2014;37:53-60.
Punaro GR, Lima DY, Rodrigues AM, Pugliero S, Mouro MG, Rogero MM, et al. Cupuacu extract reduces nitrosative stress and modulates inflammatory mediators in the kidneys of experimental diabetes. Clin Nutr. 2019;38(1):364-71.
Ceriello A. Oxidative stress and diabetes-associated complications. Endocr Pract. 2006;12 Suppl 1:60-2.
Giugliano D, Ceriello A, Paolisso G. Diabetes mellitus, hypertension, and cardiovascular disease: which role for oxidative stress? Metabolism. 1995;44(3):363-8.
Fernandes TO, Rodrigues AM, Punaro GR, Lima DY, Higa EMS. P2X7 receptor-nitric oxide interaction mediates apoptosis in mouse immortalized mesangial cells exposed to high glucose. J Bras Nefrol. 2022;44(2):147-54.
Lorenzi M, Montisano DF, Toledo S, Barrieux A. High glucose induces DNA damage in cultured human endothelial cells. J Clin Invest. 1986;77(1):322-5.
Roth T, Podesta F, Stepp MA, Boeri D, Lorenzi M. Integrin overexpression induced by high glucose and by human diabetes: potential pathway to cell dysfunction in diabetic microangiopathy. Proc Natl Acad Sci U S A. 1993;90(20):9640-4.
Wu QD, Wang JH, Fennessy F, Redmond HP, Bouchier-Hayes D. Taurine prevents high-glucose-induced human vascular endothelial cell apoptosis. Am J Physiol. 1999;277(6):C1229-38.
Cai T, Cai B. Pharmacological activities of esculin and esculetin: A review. Medicine (Baltimore). 2023;102(40):e35306.
Sarfraz I, Rasul A, Jabeen F, Younis T, Zahoor MK, Arshad M, et al. Fraxinus: A Plant with Versatile Pharmacological and Biological Activities. Evid Based Complement Alternat Med. 2017;2017:4269868.
Li CX, Li JC, Lai J, Liu Y. The pharmacological and pharmacokinetic properties of esculin: A comprehensive review. Phytother Res. 2022;36(6):2434-48.
Zhang L, Xie Q, Li X. Esculetin: A review of its pharmacology and pharmacokinetics. Phytother Res. 2022;36(1):279-98.
Serralha RS, Rodrigues IF, Bertolini A, Lima DY, Nascimento M, Mouro MG, et al. Esculin reduces P2X7 and reverses mitochondrial dysfunction in the renal cortex of diabetic rats. Life Sciences. 2020:117787.
Ju S, Tan Y, Wang Q, Zhou L, Wang K, Wen C, et al. Antioxidant and anti‑inflammatory effects of esculin and esculetin (Review). Exp Ther Med. 2024;27(6):248.
Medina ME, Galano A, Alvarez-Idaboy JR. Theoretical study on the peroxyl radicals scavenging activity of esculetin and its regeneration in aqueous solution. Physical Chemistry Chemical Physics. 2014;16(3):1197-207.
Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method to evaluate antioxidant activity. LWT - Food Science and Technology. 1995;28(1):25-30.
Chen Z, Bertin R, Froldi G. EC50 estimation of antioxidant activity in DPPH. assay using several statistical programs. Food Chem. 2013;138(1):414-20.
Pruccoli L, Morroni F, Sita G, Hrelia P, Tarozzi A. Esculetin as a Bifunctional Antioxidant Prevents and Counteracts the Oxidative Stress and Neuronal Death Induced by Amyloid Protein in SH-SY5Y Cells. Antioxidants. 2020;9(6).
Asmat U, Abad K, Ismail K. Diabetes mellitus and oxidative stress-A concise review. Saudi Pharm J. 2016;24(5):547-53.
Yan LJ. Redox imbalance stress in diabetes mellitus: Role of the polyol pathway. Animal models and experimental medicine. 2018;1(1):7-13.
Cordiano R, Di Gioacchino M, Mangifesta R, Panzera C, Gangemi S, Minciullo PL. Malondialdehyde as a Potential Oxidative Stress Marker for Allergy-Oriented Diseases: An Update. Molecules. 2023;28(16).
Ayala A, Munoz MF, Arguelles S. Lipid peroxidation: production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxidative medicine and cellular longevity. 2014;2014:360438.
Del Rio D, Stewart AJ, Pellegrini N. A review of recent studies on malondialdehyde as toxic molecule and biological marker of oxidative stress. Nutrition, Metabolism and Cardiovascular Diseases. 2005;15(4):316-28.
Lankin VZ, Tikhaze AK, Melkumyants AM. Malondialdehyde as an Important Key Factor of Molecular Mechanisms of Vascular Wall Damage under Heart Diseases Development. Int J Mol Sci. 2022;24(1).
Caldiroli A, Auxilia AM, Capuzzi E, Clerici M, Buoli M. Malondialdehyde and bipolar disorder: A short comprehensive review of available literature. J Affect Disord. 2020;274:31-7.
Shi MH, Wu Y, Li L, Cai YF, Liu M, Gao XH, et al. Meta-analysis of the association between vitiligo and the level of superoxide dismutase or malondialdehyde. Clin Exp Dermatol. 2017;42(1):21-9.
Maurya RP, Prajapat MK, Singh VP, Roy M, Todi R, Bosak S, et al. Serum Malondialdehyde as a Biomarker of Oxidative Stress in Patients with Primary Ocular Carcinoma: Impact on Response to Chemotherapy. Clin Ophthalmol. 2021;15:871-9.
Pitocco D, Zaccardi F, Di Stasio E, Romitelli F, Santini SA, Zuppi C, et al. Oxidative stress, nitric oxide, and diabetes. Rev Diabet Stud. 2010;7(1):15-25.
Armas-Padilla MC, Armas-Hernandez MJ, Sosa-Canache B, Cammarata R, Pacheco B, Guerrero J, et al. Nitric oxide and malondialdehyde in human hypertension. Am J Ther. 2007;14(2):172-6.
Cosentino F, Luscher TF. Maintenance of vascular integrity: role of nitric oxide and other bradykinin mediators. Eur Heart J. 1995;16 Suppl K:4-12.
Barbato JE, Tzeng E. Nitric oxide and arterial disease. J Vasc Surg. 2004;40(1):187-93.
Noh H, Ha H, Yu MR, Kang SW, Choi KH, Han DS, et al. High Glucose Increases Inducible NO Production in Cultured Rat Mesangial Cells : Possible Role in Fibronectin Production. Nephron. 2001;90(1):78-85.
Jomova K, Alomar SY, Alwasel SH, Nepovimova E, Kuca K, Valko M. Several lines of antioxidant defense against oxidative stress: antioxidant enzymes, nanomaterials with multiple enzyme-mimicking activities, and low-molecular-weight antioxidants. Archives of toxicology. 2024;98(5):1323-67.
Dumanovic J, Nepovimova E, Natic M, Kuca K, Jacevic V. The Significance of Reactive Oxygen Species and Antioxidant Defense System in Plants: A Concise Overview. Front Plant Sci. 2020;11:552969.
Moreira MA, Nascimento MA, Bozzo TA, Cintra A, da Silva SM, Dalboni MA, et al. Ascorbic acid reduces gentamicin-induced nephrotoxicity in rats through the control of reactive oxygen species. Clinical nutrition. 2014;33(2):296-301.
Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O. Oxidative Stress and Antioxidant Defense. World Allergy Organization Journal. 2012;5(1):9-19.
Wang K, Zhang Y, Ekunwe SIN, Yi X, Liu X, Wang H, et al. Antioxidant activity and inhibition effect on the growth of human colon carcinoma (HT-29) cells of esculetin from Cortex Fraxini. Medicinal Chemistry Research. 2011;20(7):968-74.
Lee J, Yang J, Jeon J, Sang Jeong H, Lee J, Sung J. Hepatoprotective effect of esculetin on ethanol-induced liver injury in human HepG2 cells and C57BL/6J mice. Journal of Functional Foods. 2018;40:536-43.
Weiwei T, Ting Z, Chunhua M, Hongyan L. Suppressing receptor-interacting protein 140: a new sight for esculetin to treat myocardial ischemia/reperfusion injury. RSC Advances. 2016;6(113):112117-28.
Pullaiah CP, Nelson VK, Rayapu S, G VN, Kedam T. Exploring cardioprotective potential of esculetin against isoproterenol induced myocardial toxicity in rats: in vivo and in vitro evidence. BMC Pharmacol Toxicol. 2021;22(1):43.
