Trigonelline protects against alcohol-induced brain damage by inhibition of oxidative stress, TLR4/NF-κB/proinflammatory cytokines pathway, and apoptosis

Document Type : Original Article

Authors

1 Department of Pharmacology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran

2 Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran

10.22038/ijbms.2026.94209.20306

Abstract

Objective(s): Brain injury is one of the most predominant complications following excessive alcohol consumption. Oxidative, inflammatory, and apoptotic processes are the essential mechanisms involved in alcohol-induced brain damage. Trigonelline is a natural compound that has a variety of pharmacologic activities. The present study investigated the protective effect of trigonelline in alcohol-induced brain injury and its underlying mechanisms.  
Materials and Methods: Adult male mice (C57BL/6) were exposed to binge ethanol (6 g/kg/day, by gavage) and treated with trigonelline (50 and 100 mg/kg/day, orally) for 6 days. Mice were sacrificed and the brain tissues were dissected for experimental assessments.  
Results: The results showed that trigonelline alleviated alcohol-induced locomotor impairment and brain oxidative damage by decreasing lipid peroxidation and protein oxidation. Trigonelline restored the levels of protective antioxidants (GSH, SOD, and HO-1) and reduced the levels of ICAM-1 and MPO in the brains of mice exposed to alcohol. Trigonelline significantly reduced alcohol-induced brain inflammation by the inhibition of iNOS/NO, TLR4, NF-κB, and proinflammatory cytokines (TNF-α, IL-6, IL-1β, and TGF-β1). Moreover, trigonelline treatment reduced the levels of caspase-3, cytochrome c, and TUNEL positive cells in the brains of alcohol-exposed mice. 
Conclusion: These findings suggest that trigonelline protects brain against alcohol intoxication by inhibition of oxidative and inflammatory and apoptotic responses. Therefore, trigonelline may serve as a potential therapeutic approach for the protection of brain damage associated with binge alcohol consumption.

Graphical Abstract

Trigonelline protects against alcohol-induced brain damage by inhibition of oxidative stress, TLR4/NF-κB/proinflammatory cytokines pathway, and apoptosis

Keywords

Main Subjects

References

1. Bhowmick S, Alikunju S, Muneer PMA. NADPH oxidase-induced activation of transforming growth factor-beta-1 causes neuropathy by suppressing antioxidant signaling pathways in alcohol use disorder. Neuropharmacology 2022; 213: 109136.
2. Amirshahrokhi K, Niapour A. Methylsulfonylmethane protects against ethanol-induced brain injury in mice through the inhibition of oxidative stress, proinflammatory mediators and apoptotic cell death. Int Immunopharmacol 2022; 106: 108638.
3. Loheswaran G, Barr MS, Rajji TK, Blumberger DM, Le Foll B, Daskalakis ZJ. Alcohol intoxication by binge drinking impairs neuroplasticity. Brain Stimul 2016; 9: 27-32.
4. Husain K, Mejia J, Lalla J, Kazim S. Dose response of alcohol-induced changes in BP, nitric oxide and anti-oxidants in rat plasma. Pharmacol Res 2005; 51: 337-343.
5. Finnerty N, O’Riordan SL, Klamer D, Lowry J, Pålsson E. Increased brain nitric oxide levels following ethanol administration. Nitric Oxide 2015; 47: 52-57.
6. Wang Y, Wang X, Li H, Xu M, Frank J, Luo J. Binge ethanol exposure induces endoplasmic reticulum stress in the brain of adult mice. Toxicol Appl Pharmacol 2018; 356: 172-181.
7. Naseer MI, Ullah I, Narasimhan ML, Lee HY, Bressan RA, Yoon GH, et al. Neuroprotective effect of osmotin against ethanol-induced apoptotic neurodegeneration in the developing rat brain. Cell Death Dis 2014; 5:e1150.
8. Palmer E, Tyacke R, Sastre M, Lingford-Hughes A, Nutt D, Ward RJ. Alcohol hangover: Underlying biochemical, inflammatory and neurochemical mechanisms. Alcohol Alcohol 2019; 54: 196-203.
9. Alfonso-Loeches S, Pascual-Lucas M, Blanco AM, Sanchez-Vera I, Guerri C. Pivotal role of TLR4 receptors in alcohol-induced neuroinflammation and brain damage. J Neurosci 2010; 30: 8285-8295. 
10. Collins MA, Neafsey EJ. Neuroinflammatory pathways in binge alcohol-induced neuronal degeneration: Oxidative stress cascade involving aquaporin, brain edema, and phospholipase A2 activation. Neurotox Res 2012; 21: 70-78.
11. Crews FT, Vetreno RP. Neuroimmune basis of alcoholic brain damage. Int Rev Neurobiol 2014; 118: 315-357.
12. Wang F, Li Y, Zhang YJ, Zhou Y, Li S, Li HB. Natural products for the prevention and treatment of hangover and alcohol use disorder. Molecules 2016; 21: 64.
13. Nguyen V, Taine EG, Meng D, Cui T, Tan W. Pharmacological activities, therapeutic effects, and mechanistic actions of trigonelline. Int J Mol Sci 2024; 25: 3385.
14. Liu L, Du X, Zhang Z, Zhou J. Trigonelline inhibits caspase 3 to protect β cells apoptosis in streptozotocin-induced type 1 diabetic mice. Eur J Pharmacol 2018; 836: 115-121. 
15.Zhang W, Zhang Y, Chen S, Zhang H, Yuan M, Xiao L, et al. Trigonelline, an alkaloid from Leonurus japonicus houtt., suppresses mast cell activation and OVA-induced allergic asthma. Front Pharmacol 2021; 12: 687970.
16. Membrez M, Migliavacca E, Christen S, Yaku K, Trieu J, Lee AK, et al. Trigonelline is an NAD+ precursor that improves muscle function during ageing and is reduced in human sarcopenia. Nat Metab 2024; 6: 433-447.
17. Sekhar MG, Ramudu Shanmugam K, Chakrapani IS. Trigonelline, a fenugreek bioactive compound protects Heart tissue against alcohol intoxication: An in-vivo study focusing on anti-oxidant perspective. J Ayurveda Integr Med 2024; 15: 100963.
18. Farid MM, Yang X, Kuboyama T, Tohda C. Trigonelline recovers memory function in Alzheimer’s disease model mice: Evidence of brain penetration and target molecule. Sci Rep 2020; 10: 16424.
19. Kabiri-Samani N, Amini-Khoei H, Rahimi-Madiseh M, Sureda A, Lorigooini Z. Trigonelline as an anticonvulsant agent: Mechanistic insights into NMDA receptor expression and oxidative stress balance. Sci Rep 2024; 14: 14239.
20. Khalili M, Alavi M, Esmaeil-Jamaat E, Baluchnejadmojarad T, Roghani M. Trigonelline mitigates lipopolysaccharide-induced learning and memory impairment in the rat due to its anti-oxidative and anti-inflammatory effect. Int Immunopharmacol 2018; 61: 355-362.
21. Pravalika K, Sarmah D, Kaur H, Vats K, Saraf J, Wanve M, et al. Trigonelline therapy confers neuroprotection by reduced glutathione mediated myeloperoxidase expression in animal model of ischemic stroke. Life Sci 2019; 216: 49-58.
22. Qiu Z, Ma J, Zhang X, Jiao M, Zhi L. Electroacupuncture combined with trigonelline inhibits pyroptosis in cerebral ischemia-reperfusion by suppressing autophagy via the PI3K/AKT/mTOR signaling pathway. Brain Res Bull 2025; 221: 111200. 
23. Pruett S, Tan W, Howell GE 3rd, Nanduri B. Dosage scaling of alcohol in binge exposure models in mice: An empirical assessment of the relationship between dose, alcohol exposure, and peak blood concentrations in humans and mice. Alcohol 2020; 89: 9-17. 
24. Chowdhury AA, Gawali NB, Bulani VD, Kothavade PS, Mestry SN, Deshpande PS, et al. In vitro antiglycating effect and in vivo neuroprotective activity of Trigonelline in d-galactose induced cognitive impairment. Pharmacol Rep 2018; 70: 372-377.
25. Amirshahrokhi K, Imani M. Therapeutic effect of levetiracetam against thioacetamide-induced hepatic encephalopathy through inhibition of oxidative Stress and downregulation of NF-κB, NLRP3, iNOS/NO, pro-inflammatory cytokines and apoptosis. Inflammation 2024; 47: 1762-1775.
26. Amirshahrokhi K, Imani M. Levetiracetam attenuates experimental ulcerative colitis through promoting Nrf2/HO-1 antioxidant and inhibiting NF-κB, proinflammatory cytokines and iNOS/NO pathways. Int Immunopharmacol 2023; 119: 110165.
27. Amirshahrokhi K, Imani M. Edaravone reduces brain injury in hepatic encephalopathy by upregulation of Nrf2/HO-1 and inhibition of NF-κB, iNOS/NO and inflammatory cytokines. Mol Biol Rep 2025; 52: 222.
28. Amirshahrokhi K, Niapour A. Carvedilol attenuates brain damage in mice with hepatic encephalopathy. Int Immunopharmacol 2022; 111: 109119. 
29. Karadayian AG, Malanga G, Czerniczyniec A, Lombardi P, Bustamante J, Lores-Arnaiz S. Free radical production and anti-oxidant status in brain cortex non-synaptic mitochondria and synaptosomes at alcohol hangover onset. Free Radic Biol Med 2017; 108: 692-703. 
30. Ding Y, Wu Y, Chen J, Zhou Z, Zhao B, Zhao R, et al. Protective effect of Eucommia ulmoides Oliver male flowers on ethanol-induced DNA damage in mouse cerebellum and cerebral cortex. Food Sci Nutr 2022; 10: 2794-2803.
31. Hamelink C, Hampson A, Wink DA, Eiden LE, Eskay RL. Comparison of cannabidiol, anti-oxidants, and diuretics in reversing binge ethanol-induced neurotoxicity. J Pharmacol Exp Ther 2005; 314: 780-788.
32. Qiu Z, Wang K, Jiang C, Su Y, Fan X, Li J, et al.Trigonelline protects hippocampal neurons from oxygen-glucose deprivation-induced injury through activating the PI3K/Akt pathway. Chem Biol Interact 2020; 317: 108946. 
33. Finnerty N, O’Riordan SL, Klamer D, Lowry J, Pålsson E. Increased brain nitric oxide levels following ethanol administration. Nitric Oxide. 2015; 47: 52-57.
34. Rump TJ, Muneer PMA, Szlachetka AM, Lamb A, Haorei C, Alikunju S, et al. Acetyl-L-carnitine protects neuronal function from alcohol-induced oxidative damage in the brain. Free Radic Biol Med 2010; 49: 1494-1504. 
35. Szabo G, Lippai D. Converging actions of alcohol on liver and brain immune signaling. Int Rev Neurobiol 2014; 118: 359-380. 
36. Alfonso-Loeches S, Ureña-Peralta J, Morillo-Bargues MJ, Gómez-Pinedo U, Guerri C. Ethanol-induced TLR4/NLRP3 neuroinflammatory response in microglial cells promotes leukocyte infiltration across the BBB. Neurochem Res 2016; 41: 193-209. 
37. Dietrich JB. The adhesion molecule ICAM-1 and its regulation in relation with the blood-brain barrier. J Neuroimmunol 2002; 128: 58-68. 
38. Sacanella E, Estruch R. The effect of alcohol consumption on endothelial adhesion molecule expression. Addict Biol 2003; 8: 371-378. 
39. Qin L, Crews FT. Chronic ethanol increases systemic TLR3 agonist-induced neuroinflammation and neurodegeneration. J Neuroinflammation 2012; 9: 130. 
40. Crews F, Nixon K, Kim D, Joseph J, Shukitt-Hale B, Qin L, et al. BHT blocks NF-kappaB activation and ethanol-induced brain damage. Alcohol Clin Exp Res 2006; 30(11): 1938-49. 
41. Paine A, Eiz-Vesper B, Blasczyk R, Immenschuh S. Signaling to heme oxygenase-1 and its anti-inflammatory therapeutic potential. Biochem Pharmacol 2010; 80: 1895-1903. 
42. Akamatsu Y, Pagan VA, Hanafy KA. The role of TLR4 and HO-1 in neuroinflammation after subarachnoid hemorrhage. J Neurosci Res 2020; 98: 549-556. 
43. Qin L, He J, Hanes RN, Pluzarev O, Hong JS, Crews FT. Increased systemic and brain cytokine production and neuroinflammation by endotoxin following ethanol treatment. J Neuroinflammation 2008; 5: 10.
44. Johnson S, Duncan J, Hussain SA, Chen G, Luo J, Mclaurin C, et al. The IFNγ-PKR pathway in the prefrontal cortex reactions to chronic excessive alcohol use. Alcohol Clin Exp Res 2015; 39: 476-484.  
45. Bhowmick S, Alikunju S, Muneer PMA. NADPH oxidase-induced activation of transforming growth factor-beta-1 causes neuropathy by suppressing antioxidant signaling pathways in alcohol use disorder. Neuropharmacology 2022; 213: 109136. 
46. Tiwari V, Chopra K. Attenuation of oxidative stress, neuroinflammation, and apoptosis by curcumin prevents cognitive deficits in rats postnatally exposed to ethanol. Psychopharmacology (Berl) 2012; 224: 519-535. 
47. Ikegami Y, Goodenough S, Inoue Y, Dodd PR, Wilce PA, Matsumoto I. Increased TUNEL positive cells in human alcoholic brains. Neurosci Lett 2003; 349: 201-205. 
 
Iranian Journal of Basic Medical Sciences
Volume 29, Issue 7
July 2026
Pages 1079-1088

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