The Comparasion of The Effects of Novel Antioxidant Compounds Ebselen, Isorhamnetin, and Genistein on the Cardiovascular System

Year 2025, Volume: 3 Issue: 2, 58 - 69, 31.08.2025

Onural Özhan , Ahmet Acet , Azibe Yıldız , Necip Ermiş , Burhan Ateş , Nigar Vardı , Cemil Çolak , Mehmet Gunata , Hakan Parlakpınar

https://doi.org/10.62425/rtpharma.1669358

Abstract

Purpose: This study evaluated the effects of antioxidants Ebselen (EBS), Isorhamnetin (ISO), and Genistein (GEN) on oxidative stress (OS), hemodynamic parameters, and cardiovascular histopathology in rats.

Methods: Forty adult Wistar rats were randomized into four groups (n=8/group): control (vehicle), EBS (15 mg/kg/day), ISO (5 mg/kg/day), and GEN (10 mg/kg/day) for 7 days. Hemodynamic parameters, biochemical markers, and histopathological changes in cardiac and aortic tissues were analyzed. Caspase-3 immunoreactivity assessed apoptosis.

Results: EBS significantly increased diastolic and mean blood pressure versus control (p<0.05). All treatments reduced cardiac malondialdehyde, indicating OS attenuation. GEN exhibited the strongest antioxidant effect, with improved glutathione and catalase activity. EBS paradoxically reduced aortic glutathione and increased malondialdehyde, suggesting pro-oxidant potential. Histopathology revealed myocardial degeneration in EBS and ISO groups, while GEN preserved cardiac structure. ISO and GEN enhanced aortic glutathione, but EBS showed mixed effects. Serum alanine aminotransferase elevation in ISO and GEN indicated mild hepatotoxicity. Caspase-3 immunoreactivity was elevated in EBS-treated hearts (p<0.001), implying pro-apoptotic effects.

Conclusion: GEN demonstrated robust antioxidant and cardioprotective effects, making it a promising therapeutic candidate. ISO showed vascular benefits but requires further safety evaluation. EBS, despite reducing OS, may induce adverse cardiac effects, warranting caution. These findings underscore the need for targeted antioxidant therapies in cardiovascular diseases, balancing efficacy and safety.

Ethics approval
An application was made to the Inonu University Faculty of Medicine Animal Experiments Local Ethics Committee for ethical approval, and ethics committee permission was obtained with ethical approval number 2016/A-103.

Author’s contributions
OO: Supervision, Conceptualization, Software, Visualization, Data curation, Writing- Original draft preparation, Writing-Reviewing and Editing. AA: Supervision, Writing-Reviewing and Editing. AY: Investigation, Methodology, Visualization. NE: Investigation, Methodology, Visualization. BA: Investigation, Data curation, Validation, Investigation, Methodology. NV: Investigation, Data curation, Validation, Investigation, Methodology. CC: Investigation, Methodology, Visualization. MG: Writing-Reviewing and Editing. HP: Supervise, write, review, and Edit.
Conflicts of interest/Competing interests
The authors have no relevant financial or non-financial interests to disclose.
Funding
This work was supported by Research Fund of the Inonu University. (Project number: 2016/189 belongs to Haci Ahmet Acet.).

Keywords

ebselen , genistein , cardiovascular , isorhamnetin , oxidative stress

Project Number

Research Fund of the Inonu University. (Project number: 2016/189 belongs to Haci Ahmet Acet.)

References

• Abudalo, R., Gammoh, O., Altaber, S., Bseiso, Y., Qnais, E., Wedyan, M., et al. (2024). Mitigation of cisplatin-induced cardiotoxicity by isorhamnetin: Mechanistic insights into oxidative stress, inflammation, and apoptosis modulation. Toxicology Reports, 12, 564–573.
• Aebi, H. (1984). Catalase in vitro. Methods in Enzymology, 105, 121–126.
• Ahwach, S. M., Thomas, M., Onstead-Haas, L., Mooradian, A. D., & Haas, M. J. (2015). The glutathione mimic ebselen inhibits oxidative stress but not endoplasmic reticulum stress in endothelial cells. Life Sciences, 134, 9–15.
• Amerizadeh, A., Asgary, S., Vaseghi, G., & Farajzadegan, Z. (2022). Effect of genistein intake on some cardiovascular risk factors: An updated systematic review and meta-analysis. Current Problems in Cardiology, 47(9), 100902.
• Azad, G. K., & Tomar, R. S. (2014). Ebselen, a promising antioxidant drug: Mechanisms of action and targets of biological pathways. Molecular Biology Reports, 41(8), 4865–4879.
• Basarslan, F., Yilmaz, N., Davarci, I., Akin, M., Ozgur, M., Yilmaz, C., et al. (2013). Effects of ebselen on radiocontrast media-induced hepatotoxicity in rats. Toxicology and Industrial Health, 29(8), 746–752.
• Choi, E. J., & Lee, B. H. (2004). Evidence for genistein-mediated cytotoxicity and apoptosis in rat brain. Life Sciences, 75(4), 499–509.
• Dhalla, N. S., Temsah, R. M., & Netticadan, T. (2000). Role of oxidative stress in cardiovascular diseases. Journal of Hypertension, 18(6), 655–673.
• Dubois-Deruy, E., Peugnet, V., Turkieh, A., & Pinet, F. (2020). Oxidative stress in cardiovascular diseases. Antioxidants, 9(9).
• Ellman, G. L. (2022). Reprint of: Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics, 726, 109245.
• Erel, O. (2004). A novel automated direct measurement method for total antioxidant capacity using a new generation, more stable ABTS radical cation. Clinical Biochemistry, 37(4), 277–285.
• Erel, O. (2005). A new automated colorimetric method for measuring total oxidant status. Clinical Biochemistry, 38(12), 1103–1111.
• Farías, J. G., Molina, V. M., Carrasco, R. A., Zepeda, A. B., Figueroa, E., Letelier, P., et al. (2017). Antioxidant therapeutic strategies for cardiovascular conditions associated with oxidative stress. Nutrients, 9(9).
• Farruggio, S., Raina, G., Cocomazzi, G., Librasi, C., Mary, D., Gentilli, S., et al. (2019). Genistein improves viability, proliferation and mitochondrial function of cardiomyoblasts cultured in physiologic and peroxidative conditions. International Journal of Molecular Medicine, 44(6), 2298–2310.
• Gupta, S. K., Dongare, S., Mathur, R., Mohanty, I. R., Srivastava, S., Mathur, S., et al. (2015). Genistein ameliorates cardiac inflammation and oxidative stress in streptozotocin-induced diabetic cardiomyopathy in rats. Molecular and Cellular Biochemistry, 408(1–2), 63–72.
• Hermenegildo, C., Nies, E., Monsalve, E., Puertas, F. J., Higueras, V., & Romero, F. J. (1990). Some aspects of cardiac antioxidant defence: Ebselen (PZ 51) treatment increases glutathione peroxidase activity in the rat heart. Biochemical Society Transactions, 18(6), 1193–1194.
• Jafari, S., Shoghi, M., & Khazdair, M. R. (2023). Pharmacological effects of genistein on cardiovascular diseases. Evidence-Based Complementary and Alternative Medicine, 2023, 8250219.
• Jia, Q., Wang, Y., Liu, X., Ma, S., & Yang, R. (2019). [Effects of genistein on Nrf2/HO-1 pathway in myocardial tissues of diabetic rats]. Zhong Nan Da Xue Xue Bao Yi Xue Ban, 44(8), 850–856.
• Kalayci, M., Coskun, O., Cagavi, F., Kanter, M., Armutcu, F., Gul, S., et al. (2005). Neuroprotective effects of ebselen on experimental spinal cord injury in rats. Neurochemical Research, 30(3), 403–410.
• Kibel, A., Lukinac, A. M., Dambic, V., Juric, I., & Selthofer-Relatic, K. (2020). Oxidative stress in ischemic heart disease. Oxidative Medicine and Cellular Longevity, 2020, 6627144.
• Li, L., Xue, J., Liu, R., Li, X., Lai, L., Xie, J., et al. (2017). Neuroprotective effects of genistein-3'-sodium sulfonate on focal cerebral ischemia in rats. Neuroscience Letters, 646, 43–48.
• Li, W., Chen, Z., Yan, M., He, P., Chen, Z., & Dai, H. (2016). The protective role of isorhamnetin on human brain microvascular endothelial cells from cytotoxicity induced by methylglyoxal and oxygen-glucose deprivation. Journal of Neurochemistry, 136(3), 651–659.
• Mihara, M., & Uchiyama, M. (1978). Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Analytical Biochemistry, 86(1), 271–278.
• Moya, E. A., Arias, P., Varela, C., Oyarce, M. P., Del Rio, R., & Iturriaga, R. (2016). Intermittent hypoxia-induced carotid body chemosensory potentiation and hypertension are critically dependent on peroxynitrite formation. Oxidative Medicine and Cellular Longevity, 2016, 9802136.
• National Research Council. (2011). Guide for the care and use of laboratory animals (8th ed.). National Academies Press.
• Niemann, B., Rohrbach, S., Miller, M. R., Newby, D. E., Fuster, V., & Kovacic, J. C. (2017). Oxidative stress and cardiovascular risk: Obesity, diabetes, smoking, and pollution: Part 3 of a 3-part series. Journal of the American College of Cardiology, 70(2), 230–251.
• Ozbek, E., Simsek, A., Ozbek, M., & Somay, A. (2013). Caloric restriction increases internal iliac artery and penil nitric oxide synthase expression in rat: Comparison of aged and adult rats. Archivio Italiano di Urologia, Andrologia: Organo Ufficiale della Società Italiana di Ecografia Urologica e Nefrologica, 85(3), 113–117.
• Poasakate, A., Maneesai, P., Rattanakanokchai, S., Bunbupha, S., Tong-Un, T., & Pakdeechote, P. (2021). Genistein prevents nitric oxide deficiency-induced cardiac dysfunction and remodeling in rats. Antioxidants, 10(2).
• Rotariu, D., Babes, E. E., Tit, D. M., Moisi, M., Bustea, C., Stoicescu, M., et al. (2022). Oxidative stress – Complex pathological issues concerning the hallmark of cardiovascular and metabolic disorders. Biomedicine & Pharmacotherapy, 152, 113238.
• Sack, M. N., Fyhrquist, F. Y., Saijonmaa, O. J., Fuster, V., & Kovacic, J. C. (2017). Basic biology of oxidative stress and the cardiovascular system: Part 1 of a 3-part series. Journal of the American College of Cardiology, 70(2), 196–211.
• Santofimia-Castaño, P., Salido, G. M., & González, A. (2013). Ebselen alters mitochondrial physiology and reduces viability of rat hippocampal astrocytes. DNA and Cell Biology, 32(4), 147–155.
• Squadrito, F., Marini, H., Bitto, A., Altavilla, D., Polito, F., Adamo, E. B., et al. (2013). Genistein in the metabolic syndrome: Results of a randomized clinical trial. The Journal of Clinical Endocrinology & Metabolism, 98(8), 3366–3374.
• Sun, J., Sun, G., Meng, X., Wang, H., Luo, Y., Qin, M., et al. (2013). Isorhamnetin protects against doxorubicin-induced cardiotoxicity in vivo and in vitro. PLoS ONE, 8(5), e64526.
• Sun, Y., Oberley, L. W., & Li, Y. (1988). A simple method for clinical assay of superoxide dismutase. Clinical Chemistry, 34(3), 497–500.
• Tan, M., Toplu, Y., Varan, E., Sapmaz, E., Özhan, O., Parlakpınar, H., et al. (2022). The effect of genistein on cisplatin-induced ototoxicity and oxidative stress. Brazilian Journal of Otorhinolaryngology, 88(1), 105–111.
• Walker, M. J., Curtis, M. J., Hearse, D. J., Campbell, R. W., Janse, M. J., Yellon, D. M., et al. (1988). The Lambeth Conventions: Guidelines for the study of arrhythmias in ischaemia, infarction, and reperfusion. Cardiovascular Research, 22(7), 447–455.
• Wang, W., & Kang, P. M. (2020). Oxidative stress and antioxidant treatments in cardiovascular diseases. Antioxidants, 9(12).
• Waterborg, J. H., & Matthews, H. R. (1994). The Lowry method for protein quantitation. Methods in Molecular Biology (Clifton, NJ), 32, 1–4.
• Wu, J., Feng, A., Liu, C., Zhou, W., Li, K., Liu, Y., et al. (2024). Genistein alleviates doxorubicin-induced cardiomyocyte autophagy and apoptosis via ERK/STAT3/c-Myc signaling pathway in rat model. Phytotherapy Research: PTR, 38(8), 3921–3934.
• Xu, Y., Tang, C., Tan, S., Duan, J., Tian, H., & Yang, Y. (2020). Cardioprotective effect of isorhamnetin against myocardial ischemia reperfusion (I/R) injury in isolated rat heart through attenuation of apoptosis. Journal of Cellular and Molecular Medicine, 24(11), 6253–6262.
• Yousefi, H., Karimi, P., Alihemmati, A., Alipour, M. R., Habibi, P., & Ahmadiasl, N. (2017). Therapeutic potential of genistein in ovariectomy-induced pancreatic injury in diabetic rats: The regulation of MAPK pathway and apoptosis. Iranian Journal of Basic Medical Sciences, 20(9), 1009–1015.
• Zhao, T. T., Yang, T. L., Gong, L., & Wu, P. (2018). Isorhamnetin protects against hypoxia/reoxygenation-induced injury by attenuating apoptosis and oxidative stress in H9c2 cardiomyocytes. Gene, 666, 92–99.