Sains Malaysiana 54(2)(2025): 569-575

http://doi.org/10.17576/jsm-2025-5402-21

Vitexin Promotes Nitric Oxide Production in Human Umbilical Vein Endothelial Cells

 (Viteksin Merangsang Penghasilan Nitrik Oksida dalam Sel Endotelium Vena Umbilikus Manusia)

AZIZAH UGUSMAN*, IZZAT ZULHILMI ABD RAHMAN, NINA DIYANA RUSANUAR, AMILIA AMINUDDIN & ADILA A HAMID

Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, 56000 Cheras, Kuala Lumpur, Malaysia

Received: 24 September 2024/Accepted: 11 November 2024

Abstract

Endothelial nitric oxide synthase (eNOS) plays a pivotal vasculoprotective role by synthesising nitric oxide (NO), a vital anti-atherogenic agent in the vascular system. Vitexin, a flavonoid, has gained attention for its extensive vasculoprotective effects. The aim of this study was to explore the effect of vitexin on eNOS and NO synthesis in human umbilical vein endothelial cells (HUVECs). Cultured HUVECs were assigned to four distinct groups: control, oxidative stress induced by 180 μM hydrogen peroxide (H₂O₂), treatment with 300 µM vitexin, and simultaneous treatment with H₂O₂ and vitexin. After 24 h, HUVECs were harvested and the mRNA expression, protein levels and activity of eNOS were measured. Additionally, NO levels were determined. The study findings showed that the HUVECs treated with vitexin demonstrated an enhanced NO production compared to the control group. Vitexin facilitated production of NO in HUVECs by upregulating the expression of the eNOS gene, promoting eNOS protein synthesis, and increasing eNOS activity. In conclusion, vitexin enhances endothelial function by stimulating endothelial NO production by eNOS, suggesting its potential as a therapeutic agent for vascular health.

Keywords: Endothelial nitric oxide synthase; human umbilical vein endothelial cells; nitric oxide; oxidative stress; vitexin

Abstrak

Endotelium nitrik oksida sintase (eNOS) memainkan peranan penting dalam perlindungan vaskular dengan menghasilkan nitrat oksida (NO) yang merupakan agen anti-aterogenik penting dalam sistem vaskular. Viteksin, sejenis flavonoid, telah mendapat perhatian kerana kesan perlindungan vaskularnya yang menyeluruh. Tujuan penyelidikan ini dijalankan adalah untuk mengkaji kesan viteksin terhadap sintesis eNOS dan NO dalam sel endotelium vena umbilikal manusia (HUVECs). Kultur HUVECs dibahagikan kepada empat kumpulan yang berbeza: kumpulan kawalan, tekanan oksidatif yang diaruh oleh 180 μM hidrogen peroksida (H₂O₂), rawatan dengan 300 µM viteksin dan rawatan serentak dengan H₂O₂ dan viteksin. Selepas 24 jam, HUVECs dituai dan ekspresi mRNA, paras protein dan aktiviti eNOS diukur. Selain itu paras NO juga ditentukan. Penemuan kajian menunjukkan bahawa HUVECs yang dirawat dengan viteksin menunjukkan peningkatan dalam penghasilan NO berbanding kumpulan kawalan. Viteksin membantu penghasilan NO dalam HUVECs dengan meningkatkan ekspresi gen eNOS, merangsang penghasilan protein eNOS dan meningkatkan aktiviti eNOS. Kesimpulannya, viteksin meningkatkan fungsi endotelium dengan merangsang penghasilan NO oleh eNOS, seterusnya mencadangkan potensinya sebagai agen terapeutik untuk kesihatan vaskular.

Kata kunci: Endotelium nitrik oksida sintase; nitrik oksida; sel endotelium vena umbilikus manusia; tekanan oksidatif; viteksin

REFERENCES

Aini, W., Yilmaz Ozdogan, C., Kara-Magden, G., Doger, E., Kucukoglu, V., Muezzinoglu, B., Ozbek, B., Canturk, Z., Cetinarslan, B., Tarkun, I. & Kenar, H. 2023. Oxidative stress elevates eNOS expression and VSMCs proliferation of the umbilical vein of GDM mothers. Medical Records 5(2): 269-276.

Ambrosino, P., Bachetti, T., D’Anna, S.E., Galloway, B., Bianco, A., D’Agnano, V., Papa, A., Motta, A., Perrotta, F. & Maniscalco, M. 2022. Mechanisms and clinical implications of endothelial dysfunction in arterial hypertension. Journal of Cardiovascular Development and Disease 9(5): 136.

Azmi, M.F., Aminuddin, A., Jamal, J.A., A Hamid, A. & Ugusman, A. 2021. Quantified Piper sarmentosum Roxb. leaves aqueous leaf extract and its antihypertensive effect in dexamethasone-induced hypertensive rats. Sains Malaysiana 50(1): 171-179.

Batty, M., Bennett, M.R. & Yu, E. 2022. The role of oxidative stress in atherosclerosis. Cells 11(23): 3843.

Bondonno, N.P., Liu, Y.L., Zheng, Y., Ivey, K., Willett, W.C., Stampfer, M.J., Rimm, E.B. & Cassidy, A. 2023. Change in habitual intakes of flavonoid-rich foods and mortality in US males and females. BMC Medicine 21(1): 181.

Chen, Y., Yang, J., Huang, Z., Yin, B., Umar, T., Yang, C., Zhang, X., Jing, H., Guo, S., Guo, M., Deng, G. & Qiu, C. 2022. Vitexin mitigates Staphylococcus aureus-induced mastitis via regulation of ROS/ER stress/NF-κB/MAPK pathway. Oxidative Medicine and Cellular Longevity 2022: 7977433.

Chen, Z. & Zhang, S.L. 2021. The role of flavonoids in the prevention and management of cardiovascular complications: A narrative review. Ann Palliat Med. 10(7): 8254-8263. 

Daiber, A., Xia, N., Steven, S., Oelze, M., Hanf, A., Kröller-Schön, S., Münzel, T. & Li, H. 2019. New therapeutic implications of endothelial nitric oxide synthase (eNOS) function/dysfunction in cardiovascular disease. International Journal of Molecular Sciences 20(1): 187.

Higashi, Y. 2022. Roles of oxidative stress and inflammation in vascular endothelial dysfunction-related disease. Antioxidants 11(10): 1958.

Janaszak-Jasiecka, A., Płoska, A., Wierońska, J.M., Dobrucki, L.W. & Kalinowski, L. 2023. Endothelial dysfunction due to eNOS uncoupling: Molecular mechanisms as potential therapeutic targets. Cellular and Molecular Biology Letters 28(1): 21.

Janaszak-Jasiecka, A., Siekierzycka, A., Płoska, A., Dobrucki, I.T. & Kalinowski, L. 2021. Endothelial dysfunction driven by hypoxia-The influence of oxygen deficiency on NO bioavailability. Biomolecules 11(7): 982.

Konno, T., Melo, E.P., Chambers, J.E. & Avezov, E. 2021. Intracellular sources of ROS/H₂O₂ in health and neurodegeneration: Spotlight on endoplasmic reticulum. Cells 10(2): 233.

Kunter, I., Zabib, N. & Kosar, M. 2019. A review of oxidants and antioxidants in biological systems. EMU Journal of Pharmaceutical Sciences 3(1): 64-72.

Liao, L., Gong, L., Zhou, M., Xue, X., Li, Y. & Peng, C. 2021. Leonurine ameliorates oxidative stress and insufficient angiogenesis by regulating the PI3K/Akt-eNOS signaling pathway in H₂O₂-Induced HUVECs. Oxidative Medicine and Cellular Longevity 2021: 9919466.

Nguyen, T.D., Rahman, N-T., Sessa, W.C. & Lee, M.Y. 2023. Endothelial nitric oxide synthase (eNOS) S1176 phosphorylation status governs atherosclerotic lesion formation. Frontiers in Cardiovascular Medicine 10: 1279868.

Olufunmilayo, E.O., Gerke-Duncan, M.B. & Holsinger, R.M.D. 2023. Oxidative stress and antioxidants in neurodegenerative disorders. Antioxidants 12(2): 517.

Panda, P., Verma, H.K., Lakkakula, S., Merchant, N., Kadir, F., Rahman, S., Jeffree, M.S., Lakkakula, B.V.K.S. & Rao, P.V. 2022. Biomarkers of oxidative stress tethered to cardiovascular diseases. Oxidative Medicine and Cellular Longevity 2022: 9154295.

Ranjan, R., Kishore, K., Ranjan, R., TJ, S., Jha, A.K., Ojha, B.K., Kumar, S. & Kumar, R. 2023. Nutraceutical potential of vitexin: A flavone glycoside. The Journal of Phytopharmacology 12(1): 44-50.

Ransy, C., Vaz, C., Lombès, A. & Bouillaud, F. 2020. Use of H₂O₂ to cause oxidative stress, the catalase issue. International Journal of Molecular Sciences 21(23): 9149.

Ray, A., Gulati, K. & Rehman, S. 2023. Translating nitric oxide research to therapeutics: A critical appraisal. In Nitric Oxide: From Research to Therapeutics. Advances in Biochemistry in Health and Disease, edited by Ray, A., Gulati, K. Springer, Champ. pp. 1-13.

Sundar, U.M., Ugusman, A., Chua, H.K., Latip, J. & Aminuddin, A. 2019. Piper sarmentosum promotes endothelial nitric oxide production by reducing asymmetric dimethylarginine in tumor necrosis factor-α-induced human umbilical vein endothelial cells. Frontiers in Pharmacology 10: 1033.

Tran, N., Garcia, T., Aniqa, M., Ali, S., Ally, A. & Nauli, S.M. 2022. Endothelial nitric oxide synthase (eNOS) and the cardiovascular system: in physiology and in disease states. American Journal of Biomedical Science & Research 15(2): 153-177.

Ugusman, A., Zakaria, Z., Chua, K.H., Megat Mohd Nordin, N.A. & Abdullah Mahdy, Z. 2014. Role of rutin on nitric oxide synthesis in human umbilical vein endothelial cells. The Scientific World Journal 2014: 169370.

Zhang, S., Jin, S., Zhang, S., Li, Y-Y., Wang, H., Chen, Y. & Lu, H. 2022. Vitexin protects against high glucose-induced endothelial cell apoptosis and oxidative stress via Wnt/β-catenin and Nrf2 signalling pathway. Archives of Physiology and Biochemistry 130(3): 275-284.

Zhou, F., Gu, K. & Zhou, Y. 2023. Flavonoid intake is associated with lower all-cause and disease-specific mortality: The National Health and Nutrition Examination Survey 2007-2010 and 2017-2018. Frontiers in Nutrition https://doi.org/10.3389/fnut.2023.1046998

Zhou, X., Qian, Y., Yuan, D., Feng, Q. & He, P. 2019. H₂O₂‐induced microvessel barrier dysfunction: The interplay between reactive oxygen species, nitric oxide, and peroxynitrite. Physiological Reports 7(16): phy2.14206.

*Corresponding author; email: dr.azizah@ppukm.ukm.edu.my