Alsari, A., Shaki, F., Chahardori, M. (2026). Protective Effects of Sulforaphane and Kojic Acid Against Arsenic-Induced Inflammatory and Apoptotic Toxicity in HaCaT Cells. , 23(3), 107-115. doi: 10.33899/iraqij.p.2026.171662.1208
Aisha Hussein Alsari; Fatemeh Shaki; Milad Chahardori. "Protective Effects of Sulforaphane and Kojic Acid Against Arsenic-Induced Inflammatory and Apoptotic Toxicity in HaCaT Cells". , 23, 3, 2026, 107-115. doi: 10.33899/iraqij.p.2026.171662.1208
Alsari, A., Shaki, F., Chahardori, M. (2026). 'Protective Effects of Sulforaphane and Kojic Acid Against Arsenic-Induced Inflammatory and Apoptotic Toxicity in HaCaT Cells', , 23(3), pp. 107-115. doi: 10.33899/iraqij.p.2026.171662.1208
Alsari, A., Shaki, F., Chahardori, M. Protective Effects of Sulforaphane and Kojic Acid Against Arsenic-Induced Inflammatory and Apoptotic Toxicity in HaCaT Cells. , 2026; 23(3): 107-115. doi: 10.33899/iraqij.p.2026.171662.1208

Protective Effects of Sulforaphane and Kojic Acid Against Arsenic-Induced Inflammatory and Apoptotic Toxicity in HaCaT Cells

Iraqi Journal of Pharmacy
Article 3, Volume 23, Issue 3, September 2026, Pages 107-115    PDF (892.75 K)
Document Type: Research Paper
DOI: 10.33899/iraqij.p.2026.171662.1208
Authors
Aisha Hussein Alsari; Fatemeh Shaki* ; Milad Chahardori
Department of Toxicology, College of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran
Abstract
Background: Arsenic is an environmental toxicant associated with skin injury, inflammation, and apoptosis-related cellular responses. Human keratinocyte (HaCaT) cells are widely used to investigate the mechanisms of arsenic-induced skin toxicity and potential protective interventions. Sulforaphane and kojic acid have been reported to possess cytoprotective and anti-inflammatory properties in various experimental models. Aim: To evaluate the protective effects of sulforaphane and kojic acid against arsenic-induced cytotoxicity and alterations in inflammation- and apoptosis-related gene expression in HaCaT cells. Methods: HaCaT cells were exposed to arsenic (5 μM) alone or in combination with sulforaphane (5, 10, and 20 μM) or kojic acid (50, 100, and 200 μM). Cell viability was assessed using the MTT assay. The expression of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), BCL-2-associated X protein (BAX), and B-cell lymphoma 2 (BCL-2) were evaluated by quantitative real-time polymerase chain reaction (qPCR). Results: Arsenic exposure significantly reduced cell viability and increased TNF-α, IL-6, and BAX expression while decreasing BCL-2 expression. Treatment with sulforaphane or kojic acid improved cell viability and attenuated arsenic-induced changes in gene expression. Both compounds reduced TNF-α, IL-6, and BAX expression and partially restored BCL-2 expression compared with the arsenic-treated group. Sulforaphane generally produced a more consistent protective effect than kojic acid across the evaluated inflammatory and apoptosis-related markers. Conclusion: Arsenic induced cytotoxicity and significant transcriptional alterations associated with inflammatory and apoptosis-related responses in HaCaT cells. Sulforaphane and kojic acid mitigated these effects, with sulforaphane showing a more consistent protective profile. These findings suggest that both compounds may help attenuate arsenic-induced keratinocyte injury and provide a basis for further mechanistic investigation.
Keywords
Arsenic; Apoptosis; HaCaT cells; Kojic acid; Sulforaphane
References
  1. Smith AH, Lingas EO, Rahman M. Contamination of drinking-water by arsenic in Bangladesh: a public health emergency. Bulletin of the World Health Organization 2000;78(9):1093–1103.
  2. Yu HS, Liao WT, Chai CY. Arsenic carcinogenesis in the skin. Journal of Biomedical Science 2006;13(5):657–666.
  3. Rossman TG, Uddin AN, Burns FJ. Arsenite is a cocarcinogen with solar ultraviolet radiation for mouse skin: an animal model for arsenic carcinogenesis. Toxicology and Applied Pharmacology 2001;176(1):64–71.
  4. Kochhar TS, Howard W, Hoffman S, Brammer-Carleton L. Effect of trivalent and pentavalent arsenic in causing chromosome alterations in cultured Chinese hamster ovary (CHO) cells. Toxicology Letters 1996;84(1):37–42.
  5. Germolec DR, Yoshida T, Gaido K, Wilmer JL, Simeonova PP, Kayama F, et al. Arsenic can mediate skin neoplasia by chronic stimulation of keratinocyte-derived growth factors. Mutation Research 1997;386(3):209–218.
  6. Shi H, Shi X, Liu KJ. Oxidative mechanism of arsenic toxicity and carcinogenesis. Molecular and Cellular Biochemistry 2004;255(1–2):67–78.
  7. Flora SJ. Arsenic-induced oxidative stress and its reversibility. Free Radical Biology and Medicine 2011;51(2):257–281.
  8. Hamadeh HK, Vargas M, Lee E, Menzel DB. Arsenic disrupts cellular levels of p53 and mdm2: a potential mechanism of carcinogenesis. Biochemical and Biophysical Research Communications 1999;263(2):446–449.
  9. Bunderson M, Coffin JD, Beall HD. Arsenic induces peroxynitrite generation and cyclooxygenase-2 protein expression in aortic endothelial cells: possible role in atherosclerosis. Toxicology and Applied Pharmacology 2002;184(1):11–18.
  10. Germolec DR, Yoshida T, Gaido K, Wilmer JL, Simeonova PP, Kayama F, et al. Arsenic induces overexpression of growth factors in human keratinocytes. Toxicology and Applied Pharmacology 1996;141(1):308–318.
  11. Meyer JN, Leung MC, Rooney JP, Sendoel A, Hengartner MO, Kisby GE, et al. Mitochondria as a target of environmental toxicants. Toxicological Sciences 2013;134(1):1–17.
  12. Ding W, Hudson LG, Liu KJ. Inorganic arsenic compounds cause oxidative damage to DNA and protein by inducing ROS and RNS generation in human keratinocytes. Molecular and Cellular Biochemistry 2005;279(1–2):105–112.
  13. Chung YP, Chen YW, Weng TI, Yang RS, Liu SH. Arsenic induces human chondrocyte senescence and accelerates rat articular cartilage aging. Archives of Toxicology 2020;94(1):89–101.
  14. Yamaguchi Y, Madhyastha H, Madhyastha R, Choijookhuu N, Hishikawa Y, Pengjam Y, et al. Arsenic acid inhibits proliferation of skin fibroblasts, and increases cellular senescence through ROS mediated MST1-FOXO signaling pathway. Journal of Toxicological Sciences 2016;41(1):105–113.
  15. Li T, Pang Q, Liu Y, Bai M, Peng Y, Zhang Z. Sulforaphane protects human umbilical vein endothelial cells from oxidative stress via the miR-34a/SIRT1 axis by upregulating nuclear factor erythroid-2-related factor 2. Experimental and Therapeutic Medicine 2021;21(3):186.
  16. Zhang YJ, Wu Q. Sulforaphane protects intestinal epithelial cells against lipopolysaccharide-induced injury by activating the AMPK/SIRT1/PGC-1ɑ Bioengineered 2021;12(1):4349-60.
  17. Chung JY, Chen H, Zirkin B. Sirt1 and Nrf2: regulation of Leydig cell oxidant/antioxidant intracellular environment and steroid formation. Biology of Reproduction 2021;105(5):1307-16.
  18. Li B, Kim DS, Yadav RK, Kim HR, Chae HJ. Sulforaphane prevents doxorubicin-induced oxidative stress and cell death in rat H9c2 cells. International Journal of Molecular Medicine 2015;36(1):53–64.
  19. Carrasco-Pozo C, Tan KN, Gotteland M, Borges K. Sulforaphane protects against high cholesterol-induced mitochondrial bioenergetics impairments, inflammation, and oxidative stress and preserves pancreatic β-cells function. Oxidative Medicine and Cellular Longevity 2017;2017:3839756.
  20. Chen M, Huang L, Lv Y, Li L, Dong Q. Sulforaphane protects against oxidative stress-induced apoptosis via activating SIRT1 in mouse osteoarthritis. Molecular Medicine Reports 2021;24(2):612.
  21. Yang PM, Cheng KC, Huang JY, Wang SY, Lin YN, Tseng YT, et al. Sulforaphane inhibits blue light-induced inflammation and apoptosis by upregulating the SIRT1/PGC-1α/Nrf2 pathway and autophagy in retinal pigment epithelial cells. Toxicology and Applied Pharmacology 2021;421:115545.
  22. Sakurai H, Morishima Y, Ishii Y, Yoshida K, Nakajima M, Tsunoda Y, et al. Sulforaphane ameliorates steroid insensitivity through an Nrf2-dependent pathway in cigarette smoke-exposed asthmatic mice. Free Radical Biology and Medicine 2018;129:473–485.
  23. Thangapandiyan S, Hema T, Miltonprabu S, Paulpandi M, Dutta U. Sulforaphane ameliorate Arsenic induced cardiotoxicity in rats: Role of PI3k/Akt mediated Nrf2 signaling pathway. Journal of Biochemical and Molecular Toxicology 2024 Jan;38(1):e23576.
  24. Shah A, Varma M, Bhandari R. Exploring sulforaphane as neurotherapeutic: Targeting Nrf2-Keap & Nf-Kb pathway crosstalk in ASD. Metabolic Brain Disease 2024;39(3):373-85.
  25. Kim YJ, Uyama H. Tyrosinase inhibitors from natural and synthetic sources: structure, inhibition mechanism and perspective for the future. Cellular and Molecular Life Sciences 2005;62(15):1707–1723.
  26. Parvez S, Kang M, Chung HS, Bae H. Naturally occurring tyrosinase inhibitors: mechanism and applications in skin health, cosmetics and agriculture industries. Phytotherapy Research 2007;21(9):805–816.
  27. Lajis AF, Hamid M, Ariff AB. Depigmenting effect of kojic acid esters in hyperpigmented B16F1 melanoma cells. Journal of Biomedicine and Biotechnology 2012;2012:952452.
  28. Khezri K, Saeedi M, Morteza-Semnani K, Akbari J, Hedayatizadeh-Omran A. A promising and effective platform for delivering hydrophilic depigmenting agents in the treatment of cutaneous hyperpigmentation: kojic acid nanostructured lipid carrier. Artificial Cells, Nanomedicine, and Biotechnology 2021;49(1):38–47.
  29. Suthiram KT, Ghazi T, Abdul NS, Chuturgoon AA. Kojic acid induces oxidative stress and anti-inflammatory effects in human hepatocellular carcinoma (HepG2) cells. Toxicon 2023;232:107221.
  30. Ali W, Choe K, Park JS, Ahmad R, Park HY, et al. Kojic acid reverses LPS-induced neuroinflammation and cognitive impairment by regulating the TLR4/NF-κB signaling pathway. Frontiers in Pharmacology 2024;15:1443552.
  31. Li TX, Liang JX, Liu LL, Shi FC, Jia XW, Li MH, et al. Novel kojic acid derivatives with anti-inflammatory effects from Aspergillus versicolor. Fitoterapia 2021;154:105027.
  32. Khan A, Park TJ, Ikram M, Ahmad S, Ahmad R, Jo MG, et al. Antioxidative and anti-inflammatory effects of kojic acid in Aβ-induced mouse model of Alzheimer's disease. Molecular Neurobiology 2021;58(10):5127–5140.
  33. Da Costa JP, Rodrigues APD, Farias LHS, Frade PCR, Da Silva BJM, Do Nascimento JLM, et al. Biological effects of kojic acid on human monocytes in vitro. Biomedicine & Pharmacotherapy 2018;101:100–106.
  34. Ram D, Karthikeyan C, Soni P, Moorthy NH. The Therapeutic Promise of Kojic Acid: A Comprehensive Review. Mini-Reviews in Medicinal Chemistry 2026. https://doi.org/10.2174/0113895575405163251027232908 
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