Examining the role of trans-sodium crocetinate in alleviating colistin-induced cytotoxicity through apoptosis and autophagy pathways on HEK-293 cells

Articles in Press

Document Type : Original Article

Authors

1 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

3 Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Physiology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

5 Applied biomedical Research Center, Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran

6 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran 2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran 3 Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran 4 Department of Physiology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran 5 Applied biomedical Research Center, Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran

10.22038/ijbms.2025.82430.17820

Abstract

Objective(s): Colistin is a crucial antibiotic for multidrug-resistant Gram-negative bacterial infections, but its nephrotoxicity limits clinical use. Trans sodium crocetinate (TSC), a synthetic crocetin derivative, exhibits anti-oxidative, antiapoptotic, and renal-protective effects. This study investigated whether TSC could alleviate colistin-induced cytotoxicity in HEK-293 cells, a human renal epithelial model.
Materials and Methods: HEK-293 cells were pretreated with varying TSC concentrations for 24 hr, followed by 200 µM colistin for another 24 hr. Cell viability was measured via MTT assay, and reactive oxygen species (ROS) levels were quantified using DCFH-DA fluorescence. Apoptotic markers (Bax, Bcl-2, caspase-3) and autophagy-related proteins (LC3, Beclin-1) were analyzed by western blotting.
Results: Colistin reduced HEK-293 cell viability by 50%, increased ROS by 43%, and elevated autophagy markers (LC3, Beclin-1) by 50%. The Bax/Bcl-2 ratio rose by 50%, and cleaved caspase-3 increased by 33% compared to controls. However, TSC pretreatment significantly attenuated these effects: viability improved by 35%, ROS decreased by 50%, and the Bax/Bcl-2 ratio dropped by 50%. Additionally, TSC reduced Bax (40%), cleaved caspase-3 (55%), LC3 (35%), and Beclin-1 (45%) levels compared to colistin-only treatment.
Conclusion: These findings suggest that TSC protects HEK-293 cells from colistin-induced toxicity by reducing oxidative stress, suppressing apoptosis, and modulating autophagy. Thus, TSC may serve as a potential adjunct therapy to mitigate colistin-associated nephrotoxicity.

Keywords

Main Subjects

References

1. Rajabalizadeh R, Ghasemzadeh Rahbardar M, Razavi BM, Hosseinzadeh H. Renoprotective effects of crocin against colistin-induced nephrotoxicity in a rat model. Iran J Basic Med Sci 2024; 27:151-156.
2. Gai Z, Samodelov SL, Kullak-Ublick GA, Visentin M. Molecular mechanisms of colistin-induced nephrotoxicity. Molecules 2019; 24:653-666.
3. Edrees NE, Galal AAA, Abdel Monaem AR, Beheiry RR, Metwally MMM. Curcumin alleviates colistin-induced nephrotoxicity and neurotoxicity in rats via attenuation of oxidative stress, inflammation and apoptosis. Chem Biol Interact 2018; 294:56-64.
4. Hanedan B, Ozkaraca M, Kirbas A, Kandemir FM, Aktas MS, Kilic K, et al. Investigation of the effects of hesperidin and chrysin on renal injury induced by colistin in rats. Biomed Pharmacother 2018; 108:1607-1616.
5. Xia C, Wang J, Wu Z, Miao Y, Chen C, Li R, Li J, Xing H. METTL3-mediated M6A methylation modification is involved in colistin-induced nephrotoxicity through apoptosis mediated by Keap1/Nrf2 signaling pathway. Toxicology 2021; 462:152961.
6. Gergin Ö, Pehlivan SS, Ulger M, Mat OC, Bayram A, Gönen ZB, et al. Efficacy of stem cell-based therapies for colistin-induced nephrotoxicity. Environ Toxicol Pharmacol 2022; 94:103933.
7. Hosseini A, Razavi BM, Hosseinzadeh H. Pharmacokinetic Properties of Saffron and its Active Components. Eur J Drug Metab Pharmacokinet 2018; 43:383-390.
8. Ghasemzadeh Rahbardar M, Hosseinzadeh H. A review of how the saffron (Crocus sativus) petal and its main constituents interact with the Nrf2 and NF-κB signaling pathways. Naunyn Schmiedebergs Arch Pharmacol 2023; 396:1879-1909.
9. Zilaee M, Hosseini SA, Jafarirad S, Abolnezhadian F, Cheraghian B, Namjoyan F, Ghadiri A. An evaluation of the effects of saffron supplementation on the asthma clinical symptoms and asthma severity in patients with mild and moderate persistent allergic asthma: A double-blind, randomized placebo-controlled trial. Respir Res 2019; 20:39.
10. Vafaeipour Z, Ghasemzadeh Rahbardar M, Hosseinzadeh H. Effect of saffron, black seed, and their main constituents on inflammatory cytokine response (mainly TNF-α) and oxidative stress status: an aspect on pharmacological insights. Naunyn Schmiedebergs Arch Pharmacol 2023; 396:2241-2259.
11. Zeinali M, Zirak MR, Rezaee SA, Karimi G, Hosseinzadeh H. Immunoregulatory and anti-inflammatory properties of Crocus sativus (Saffron) and its main active constituents: A review. Iran J Basic Med Sci 2019; 22:334-344.
12. Boskabady MH, Rahbardar MG, Jafari Z. The effect of safranal on histamine (H(1)) receptors of guinea pig tracheal chains. Fitoterapia 2011; 82:162-167.
13. Pradhan J, Mohanty C, Sahoo SK. Protective efficacy of crocetin and its nanoformulation against cyclosporine A-mediated toxicity in human embryonic kidney cells. Life Sci 2019; 216:39-48.
14. Tsiogkas SG, Grammatikopoulou MG, Gkiouras K, Zafiriou E, Papadopoulos I, Liaskos C, et al.  Effect of Crocus sativus (Saffron) intake on top of standard treatment, on disease outcomes and comorbidities in patients with rheumatic diseases: synthesis without meta-analysis (SWiM) and level of adherence to the CONSORT statement for randomized controlled trials delivering herbal medicine interventions. Nutrients 2021; 13:4274-4292.
15. Rahbardar MG, Hosseinzadeh H. 2021. Mechanisms of action of herbal antidepressants. In the Neuroscience of Depression. Eds. Martin, C.R., Hunter, L-A., Patel, V.B., Preedy, V. R. Rajendram, R. 1st Ed. Chp. 47, London: Academic Press, An Imprinted by Elsevier.
16. Xiang M, Yang M, Zhou C, Liu J, Li W, Qian Z. Crocetin prevents AGEs-induced vascular endothelial cell apoptosis. Pharmacol Res 2006; 54:268-274.
17. Rajabian F, Mehri S, Razavi BM, Khajavi Rad A, Ghasemzadeh Rahbardar M, Hosseinzadeh H. Effect of trans-sodium crocetinate on contrast-induced cytotoxicity in HEK-293 cells. Iran J Basic Med Sci 2023; 26:148-156.
18. Zhang J, Wang Y, Dong X, Liu J. Crocetin attenuates inflammation and amyloid-β accumulation in APPsw transgenic mice. Immun Ageing 2018; 15:24.
19. Nassiri-Asl M, Hosseinzadeh, H. 2020. The role of saffron and its main components on oxidative stress in neurological diseases: A review. In Oxidative stress and dietary antioxidants in neurological diseases. Eds. Martin CR, Preedy VR. 1st Ed. Chp.23, London: Academic Press, An Imprinted by Elsevier.
20. Lapchak PA. Efficacy and safety profile of the carotenoid trans sodium crocetinate administered to rabbits following multiple infarct ischemic strokes: a combination therapy study with tissue plasminogen activator. Brain Res 2010; 1309:136-145.
21. Rameshrad M, Razavi BM, Hosseinzadeh H. Saffron and its derivatives, crocin, crocetin and safranal: a patent review. Expert Opin Ther Pat 2018; 28:147-165.
22. Quintavalle C, Brenca M, De Micco F, Fiore D, Romano S, Romano MF, Apone F, Bianco A, Zabatta MA, Troncone G, Briguori C, Condorelli G. In vivo and in vitro assessment of pathways involved in contrast media-induced renal cells apoptosis. Cell Death Dis 2011; 2:e155.
23. Koc E, Reis KA, Ebinc FA, Pasaoglu H, Demirtas C, Omeroglu S, et al. Protective effect of beta-glucan on contrast induced-nephropathy and a comparison of beta-glucan with nebivolol and N-acetylcysteine in rats. Clin Exp Nephrol 2011; 15:658-665.
24. Pereira CV, Nadanaciva S, Oliveira PJ, Will Y. The contribution of oxidative stress to drug-induced organ toxicity and its detection in vitro and in vivo. Expert Opin Drug Metab Toxicol 2012; 8:219-237.
25. Deavall DG, Martin EA, Horner JM, Roberts R. Drug-induced oxidative stress and toxicity. J Toxicol 2012; 2012:645460.
26. Biswas SK. Does the interdependence between oxidative stress and inflammation explain the antioxidant paradox? Oxid Med Cell Longev 2016; 2016:5698931.
27.Mirjalili M, Mirzaei E, Vazin A. Pharmacological agents for the prevention of colistin-induced nephrotoxicity. Eur J Med Res 2022; 27:64-82.
28. Dai C, Li J, Tang S, Li J, Xiao X. Colistin-induced nephrotoxicity in mice involves the mitochondrial, death receptor, and endoplasmic reticulum pathways. Antimicrob Agents Chemother 2014; 58:4075-4085.
29. Zhao Z, Li J, Zheng B, Liang Y, Shi J, Zhang J, et al. Ameliorative effects and mechanism of crocetin in arsenic trioxide‑induced cardiotoxicity in rats. Mol Med Rep 2020; 22:5271-5281.
30. Ohba T, Ishisaka M, Tsujii S, Tsuruma K, Shimazawa M, Kubo K, et al.  Crocetin protects ultraviolet A-induced oxidative stress and cell death in skin in vitro and in vivo. Eur J Pharmacol 2016; 789:244-253.
31. Zhang L, Xie D, Chen X, Hughes ML, Jiang G, Lu Z, et al. p53 Mediates Colistin-Induced Autophagy and Apoptosis in PC-12 Cells. Antimicrob Agents Chemother 2016; 60:5294-5301.
32. Nishida K, Yamaguchi O, Otsu K. Crosstalk between autophagy and apoptosis in heart disease. Circ Res 2008; 103:343-351.
33. Zhou F, Yang Y, Xing D. Bcl-2 and Bcl-xL play important roles in the crosstalk between autophagy and apoptosis. Febs j 2011; 278:403-413.
34. Chien CT, Shyue SK, Lai MK. Bcl-xL augmentation potentially reduces ischemia/reperfusion induced proximal and distal tubular apoptosis and autophagy. Transplantation 2007; 84:1183-1190.
35. Fan YJ, Zong WX. The cellular decision between apoptosis and autophagy. Chin J Cancer 2013; 32:121-129.
36. Zhang L, Zhao Y, Ding W, Jiang G, Lu Z, Li L, Wang J, Li J, Li J.  Autophagy regulates colistin-induced apoptosis in PC-12 cells. Antimicrob Agents Chemother 2015; 59:2189-2197.
37. Zhang A, Li J. Crocetin shifts autophagic cell survival to death of breast cancer cells in chemotherapy. Tumour Biol 2017; 39:1010428317694536.
38. Wani A, Al Rihani SB, Sharma A, Weadick B, Govindarajan R, Khan SU, et al. Crocetin promotes clearance of amyloid-β by inducing autophagy via the STK11/LKB1-mediated AMPK pathway. Autophagy 2021; 17:3813-3832.
Iranian Journal of Basic Medical Sciences

Articles in Press, Accepted Manuscript
Available Online from 07 July 2025

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