Curcumin Attenuates Hippuric Acid-Induced Eryptosis by Enhancing Antioxidant Enzyme Activity

Authors

  • Namra Butt Department of Biochemistry, University of Agriculture Faisalabad, Pakistan
  • Sadia Nasir Department of Biochemistry, University of Agriculture Faisalabad, Pakistan
  • Urwah Ahmad Department of Biochemistry, University of Agriculture Faisalabad, Pakistan
  • Masooma Haider Institute of Biological Science and Technology, China Medical University, Taiwan
  • Imrana Haider Department of Biochemistry, Government College University Faisalabad, Pakistan
  • Alina Murtaza Department of Biochemistry, Quaid i Azam University Islamabad, Pakistan
  • Zara Javaid Department of Biochemistry, University of Agriculture Faisalabad, Pakistan
  • Muhammad Najamul Hassan Department of Biochemistry, University of Agriculture Faisalabad, Pakistan
  • Kashif Jilani Department of Biochemistry, University of Agriculture Faisalabad, Pakistan

DOI:

https://doi.org/10.70749/ijbr.v3i8.2044

Keywords:

Curcumin, Hippuric acid, Eryptosis, Oxidative stress, Antioxidant enzymes, Calcium signaling, Hemolysis, Chronic kidney disease

Abstract

Background: Eryptosis, the programmed death of erythrocytes, is exacerbated under pathological conditions such as chronic kidney disease (CKD), largely due to the accumulation of uremic toxins like hippuric acid. These toxins promote oxidative stress and disrupt erythrocyte membrane integrity, leading to hemolysis and anemia. Curcumin, a polyphenolic compound derived from Curcuma longa, is well known for its potent antioxidant and cyto-protective properties, yet its protective role against uremic toxin-induced eryptosis remains underexplored. Objective: This study aimed to investigate the protective effects of curcumin against hippuric acid-induced eryptosis in human erythrocytes and to explore the involvement of oxidative stress and calcium influx in the onset of eryptosis. Methods: Human erythrocytes were treated with different concentrations of hippuric acid (300 -430 μM) and curcumin (5-15 μM) for 48 hours. The hemolysis % was determined to assess the cytotoxic effect of hippuric acid. The oxidative potential of hippuric acid and antioxidant potential of curcumin was determining by assessing the activities of antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase. Morphological changes were assessed via mean corpuscular volume (MCV). Amlodipine was used to confirm the role of calcium in eryptosis.  Results: Hippuric acid significantly increased hemolysis %, reduced MCV, and suppressed activity of antioxidant enzymes (SOD, GPx, and CAT). Curcumin co-treatment markedly attenuated hemolysis %, restored MCV, and significantly upregulated antioxidant enzyme activity. Amlodipine pretreatment effectively blocked calcium channels thus mitigated the effect of hippuric acid. Conclusion: Curcumin protects erythrocytes from hippuric acid-induced eryptosis by restoring antioxidant defenses and providing cyto-protective properties. These findings reveal the protective mechanisms of curcumin and suggest its potential utility as a natural therapeutic agent to mitigate erythrocyte damage in CKD and related oxidative stress conditions.

Downloads

Download data is not yet available.

References

1. Barbalato, L. and L.S. Pillarisetty, Histology, red blood cell. StatPearls [internet], 2022.

2. Farag, M.R. and M. Alagawany, Erythrocytes as a biological model for screening of xenobiotics toxicity. Chemico-biological interactions, 2018. 279: p. 73-83.

https://doi.org/10.1016/j.cbi.2017.11.007

3. De Oliveira, S., A.S. Silva-Herdade, and C. Saldanha, Modulation of erythrocyte deformability by PKC activity. Clinical hemorheology and microcirculation, 2008. 39(1-4): p. 363-373.

https://doi.org/10.3233/ch-2008-1101

4. Kuhn, V., et al., Red blood cell function and dysfunction: redox regulation, nitric oxide metabolism, anemia. Antioxidants & redox signaling, 2017. 26(13): p. 718-742.

https://doi.org/10.1089/ars.2016.6954

5. Keohane, E.M., L. Smith, and J.M. Walenga, Rodak's hematology: clinical principles and applications. 2015: Elsevier Health Sciences.

6. Zivot, A., et al., Erythropoiesis: insights into pathophysiology and treatments in 2017. Molecular Medicine, 2018. 24(1): p. 11.

https://doi.org/10.1186/s10020-018-0011-z

7. Paulson, R.F., S. Hariharan, and J.A. Little, Stress erythropoiesis: definitions and models for its study. Experimental hematology, 2020. 89: p. 43-54. e2.

https://doi.org/10.1186/s10020-018-0011-z

8. Gottlieb, Y., et al., Physiologically aged red blood cells undergo erythrophagocytosis in vivo but not in vitro. haematologica, 2012. 97(7): p. 994.

https://doi.org/10.3324/haematol.2011.057620

9. Lang, F. and S.M. Qadri, Mechanisms and significance of eryptosis, the suicidal death of erythrocytes. Blood purification, 2012. 33(1-3): p. 125-130.

https://doi.org/10.1159/000334163

10. Biswas, D., G. Sen, and T. Biswas, Reduced cellular redox status induces 4-hydroxynonenal-mediated caspase 3 activation leading to erythrocyte death during chronic arsenic exposure in rats. Toxicology and applied pharmacology, 2010. 244(3): p. 315-327.

https://doi.org/10.1016/j.taap.2010.01.009

11. Flohé, L., Looking back at the early stages of redox biology. Antioxidants, 2020. 9(12): p. 1254.

12. Rasheed, M., et al., Endocrine Crosstalk: Diabetes as a Catalyst for Cancer Progression and Metastasis.

13. Daenen, K., et al., Oxidative stress in chronic kidney disease. Pediatric nephrology, 2019. 34(6): p. 975-991.

https://doi.org/10.1007/s00467-018-4005-4

14. Li, L., et al., Periodontitis exacerbates and promotes the progression of chronic kidney disease through oral flora, cytokines, and oxidative stress. Frontiers in microbiology, 2021. 12: p. 656372.

https://doi.org/10.3389/fmicb.2021.656372

15. Wafa, T., et al., Subacute effects of 2, 4-dichlorophenoxyacetic herbicide on antioxidant defense system and lipid peroxidation in rat erythrocytes. Pesticide Biochemistry and Physiology, 2011. 99(3): p. 256-264.

https://doi.org/10.1016/j.pestbp.2011.01.004

16. Sikora, J., et al., Hemolysis is a primary ATP-release mechanism in human erythrocytes. Blood, The Journal of the American Society of Hematology, 2014. 124(13): p. 2150-2157.

https://doi.org/10.1182/blood-2014-05-572024

17. Sun, B., et al., Hippuric acid promotes renal fibrosis by disrupting redox homeostasis via facilitation of NRF2–KEAP1–CUL3 interactions in chronic kidney disease. Antioxidants, 2020. 9(9): p. 783.

https://doi.org/10.3390/antiox9090783

18. Lim, Y.J., et al., Uremic toxins in the progression of chronic kidney disease and cardiovascular disease: mechanisms and therapeutic targets. Toxins, 2021. 13(2): p. 142.

19. Pieniazek, A., J. Bernasinska-Slomczewska, and L. Gwozdzinski, Uremic toxins and their relation with oxidative stress induced in patients with CKD. International journal of molecular sciences, 2021. 22(12): p. 6196.

https://doi.org/10.3390/ijms22126196

20. Shahidi, F. and P. Ambigaipalan, Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects–A review. Journal of functional foods, 2015. 18: p. 820-897.

https://doi.org/10.1016/j.jff.2015.06.018

21. Çimen, M.B., Free radical metabolism in human erythrocytes. Clinica chimica acta, 2008. 390(1-2): p. 1-11.

22. Remigante, A., R. Morabito, and A. Marino, Band 3 protein function and oxidative stress in erythrocytes. Journal of cellular physiology, 2021. 236(9): p. 6225-6234.

https://doi.org/10.1002/jcp.30322

23. Yang, W., et al., Effects of three kinds of curcuminoids on anti-oxidative system and membrane deformation of human peripheral blood erythrocytes in high glucose levels. Cellular Physiology and Biochemistry, 2015. 35(2): p. 789-802.

24. Barzegar, A. and A.A. Moosavi-Movahedi, Intracellular ROS protection efficiency and free radical-scavenging activity of curcumin. PloS one, 2011. 6(10): p. e26012.

https://doi.org/10.1371/journal.pone.0026012

25. Rapa, S.F., et al., Inflammation and oxidative stress in chronic kidney disease—potential therapeutic role of minerals, vitamins and plant-derived metabolites. International journal of molecular sciences, 2019. 21(1): p. 263.

https://doi.org/10.3390/ijms21010263

26. Manabe, I., Chronic inflammation links cardiovascular, metabolic and renal diseases. Circulation Journal, 2011. 75(12): p. 2739-2748.

27. Bugyei-Twum, A., et al., Suppression of NLRP3 inflammasome activation ameliorates chronic kidney disease-induced cardiac fibrosis and diastolic dysfunction. Scientific reports, 2016. 6(1): p. 39551.

https://doi.org/10.1038/srep39551

28. Bentzen, P., E. Lang, and F. Lang, Curcumin induced suicidal erythrocyte death. Cellular Physiology and Biochemistry, 2007. 19(1-4): p. 153-164.

29. de Almeida Alvarenga, L., et al., Curcumin-A promising nutritional strategy for chronic kidney disease patients. Journal of Functional Foods, 2018. 40: p. 715-721.

https://doi.org/10.1016/j.jff.2017.12.015

30. Sattar, T., et al., Induction of erythrocyte membrane blebbing by methotrexate-induced oxidative stress. Dose-Response, 2022. 20(2): p. 15593258221093853.

https://doi.org/10.1177/15593258221093853

31. Mukhtar, F., et al., Stimulation of erythrocyte membrane blebbing by bifenthrin induced oxidative stress. Dose-Response, 2022. 20(1): p. 15593258221076710.

32. Banerjee, A., et al., Concentration dependent antioxidant/pro-oxidant activity of curcumin: Studies from AAPH induced hemolysis of RBCs. Chemico-biological interactions, 2008. 174(2): p. 134-139.

https://doi.org/10.1016/j.cbi.2008.05.009

33. Zhang, J., et al., Assessment of free radicals scavenging activity of seven natural pigments and protective effects in AAPH-challenged chicken erythrocytes. Food Chemistry, 2014. 145: p. 57-65.

34. Attia, A.M., et al., Antioxidant effects of curcumin against cadmium chloride-induced oxidative stress in the blood of rats. Journal of Pharmacognosy and Phytotherapy, 2014. 6(3): p. 33-40.

https://doi.org/10.5897/jpp2014.0316

35. Verma, A., et al., Evaluation of oxidative stress and toluene exposure among adolescent inhalant users at a tertiary care Centre of North India. Free radical biology and medicine, 2017. 112: p. 37.

36. El‐Bahr, S., Effect of curcumin on hepatic antioxidant enzymes activities and gene expressions in rats intoxicated with aflatoxin B1. Phytotherapy research, 2015. 29(1): p. 134-140.

https://doi.org/10.1002/ptr.5239

37. Ming, J., et al., Optimal dietary curcumin improved growth performance, and modulated innate immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp (Ctenopharyngodon idella) after infection with Aeromonas hydrophila. Fish & shellfish immunology, 2020. 97: p. 540-553.

https://doi.org/10.1016/j.fsi.2019.12.074

38. Dehghan Haghighi, J., M. Hormozi, and A. Payandeh, Blood serum levels of selected biomarkers of oxidative stress among printing workers occupationally exposed to low-levels of toluene and xylene. Toxicology and Industrial Health, 2022. 38(5): p. 299-307.

https://doi.org/10.1177/07482337221092501

39. Awodele, O., et al., Trace elements and oxidative stress levels in the blood of painters in Lagos, Nigeria: occupational survey and health concern. Biological trace element research, 2013. 153(1): p. 127-133.

40. Özakıncı, O.G., et al., Dynamic thiol disulfide homeostasis in painters as indices of oxidative stress. International Journal of Environmental Health Research, 2022. 32(5): p. 1067-1075.

https://doi.org/10.1080/09603123.2020.1827227

41. Storka, A., et al., Effect of liposomal curcumin on red blood cells in vitro. Anticancer research, 2013. 33(9): p. 3629-3634.

42. Uluışık, D., E. Keskin, and D. Hatipoğlu, Effects of curcumin on hematological parameters in aflatoxin B1 applied rats. Turkish Journal of Sport and Exercise, 2020. 22(2): p. 265-270.

43. Rana, R.B., et al., Atorvastatin induced erythrocytes membrane blebbing. Dose-Response, 2019. 17(3): p. 1559325819869076.

https://doi.org/10.1177/1559325819869076

44. Naveed, A., et al., Induction of erythrocyte shrinkage by omeprazole. Dose-response, 2020. 18(3): p. 1559325820946941.

https://doi.org/10.1177/1559325820946941

45. Mineral Pitch Attenuates Oxidative Stress-Induced Eryptosis in Human Erythrocytes via Antioxidants and Calcium-Modulatory Mechanism. Indus Journal of Bioscience Research, 2025. 3(6): p. 520-529.

https://doi.org/10.70749/ijbr.v3i6.1690

Downloads

Published

2025-08-10

Issue

Vol. 3 No. 8 (2025): IJBR

Section

Original Article

License

Copyright (c) 2025 Indus Journal of Bioscience Research

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite

Butt, N., Nasir, S., Ahmad, U., Haider, M., Haider, I., Murtaza, A., Javaid, Z., Hassan, M. N., & Jilani, K. (2025). Curcumin Attenuates Hippuric Acid-Induced Eryptosis by Enhancing Antioxidant Enzyme Activity. Indus Journal of Bioscience Research, 3(8), 139-147. https://doi.org/10.70749/ijbr.v3i8.2044