Effect of crocin, an active saffron constituent, on ethanol toxicity in the rat: histopathological and biochemical studies

Document Type: Original Article


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

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

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

4 Orthopedic Research Center, Shahid Kamyab Hospital, Mashhad University of Medical Sciences, Mashhad, Iran

5 Ghaem Hospital, Department of Pathology, Mashhad University of Medical Sciences, Mashhad, Iran


Objective(s): The drinking of ethanol causes the wide range of clinical illness and morphological changes including hepatotoxicity and nephrotoxicity. In the current study, the protective properties of crocin versus oxidative stress, apoptosis and inflammation induced by ethanol were assessed.
Materials and Methods: The male Wistar rats were divided into seven groups consisting of 6 rats in control, ethanol (50% v/v - 5 g/kg), crocin (10, 20 and 40 mg/Kg) plus ethanol, crocin 20 and 40 mg/Kg.
Results: The MDA level was remarkably enhanced, while the content of GSH was significantly diminished in the kidney and liver of alcoholic rat but protective groups restored the level of MDA and GSH contents. Ethanol consumption induced hepatotoxicity and nephrotoxicity as evidenced by biochemical abnormalities and histopathological damages but crocin improved them. Also, crocin restored the TNF-α and IL-6 levels in the liver. The consumption of ethanol enhanced the levels of caspase-3, -8, -9 and Bax/Bcl-2 ratio (mRNA and protein) but, western blot and real-time PCR data confirmed that crocin treatment prevented apoptosis induced by ethanol.
Conclusion: This research demonstrates that crocin has protective activities against ethanol toxic effects in rat liver and kidney via anti-inflammatory, antioxidant, and anti-apoptotic effects.


1. Saranraj P, Sivasakthivelan P, Naveen M. Fermentation of fruit wine and its quality analysis: a review. AJST 2017;1:85-97.
2. Pari L, Suresh A. Effect of grape (Vitis vinifera L.) leaf extract on alcohol induced oxidative stress in rats. Food Chem Toxicol 2008;46:1627-1634.
3. Tahir M, Sultana S. Chrysin modulates ethanol metabolism in Wistar rats: a promising role against organ toxicities. Alcohol Alcohol 2011;46:383-392.
4. Faremi TY, Suru SM, Fafunso MA, Obioha UE. Hepatoprotective potentials of Phyllanthus amarus against ethanol-induced oxidative stress in rats. Food Chem Toxicol 2008;46:2658-2664.
5. Ojeda M, Barrero M, Nogales F, Murillo M, Carreras O. Oxidative effects of chronic ethanol consumption on the functions of heart and kidney: folic acid supplementation. Alcohol Alcohol 2012;47:404-412.
6. Kode A, Rajagopalan R, Penumastha SV, Menon VP. Role of an aminothiazole derivative on ethanol-induced toxicity. Toxicol Mech Methods 2007;17:33-40.
7. Saravanan R, Pugalendi K. Effect of Piper betel on blood glucose and lipid profiles in rats after chronic ethanol administration. Pharm Biol 2004;42:323-327.
8. Mani V, Siddique AI, Arivalagan S, Thomas NS, Namasivayam N. Zingerone ameliorates hepatic and renal damage in alcohol-induced toxicity in experimental rats. Int J Nutr Pharmacol Neurol Dis 2016;6:125-132.
9. You Y, Yoo S, Yoon HG, Park J, Lee YH, Kim S, et al. In vitro and in vivo hepatoprotective effects of the aqueous extract from Taraxacum officinale (dandelion) root against alcohol-induced oxidative stress. Food Chem Toxicol 2010;48:1632-1637.
10. McVicker BL, Tuma DJ, Casey CA. Effect of ethanol on pro-apoptotic mechanisms in polarized hepatic cells. World J Gastroenterol 2007;13:4960-4966.
11. Navasumrit P, Ward TH, Dodd NJ, O’Connor PJ. Ethanol-induced free radicals and hepatic DNA strand breaks are prevented in vivo by antioxidants: effects of acute and chronic ethanol exposure. J Carcinog 2000;21:93-99.
12. Rezaee-Khorasany AR, Hosseinzadeh H. Therapeutic effects of saffron (Crocus sativus L.) in digestive disorders: a review. Iran J Basic Med Sci 2016;19:455-469.
13. Mohammadzadeh L, Abnous K, Razavi BM, Hosseinzadeh H. Crocin-protected malathion-induced spatial memory deficits by inhibiting TAU protein hyperphosphorylation and antiapoptotic effects. Nutr Neurosci 2019;21:1-16.
14. Adabizadeh M, Mehri S, Rajabpour M, Abnous K, Rashedinia M, Hosseinzadeh H. The effects of crocin on spatial memory impairment induced by hyoscine: Role of NMDA, AMPA, ERK, and CaMKII proteins in rat hippocampus. Iran J Basic Med Sci 2019;22:601-609.
15. Alavizadeh SH, Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: a comprehensive review. Food Chem Toxicol 2014;64:65-80.
16. Amin B, Nakhsaz A, Hosseinzadeh H. Evaluation of the antidepressant-like effects of acute and sub-acute administration of crocin and crocetin in mice. Avicenna J Phytomed 2015;5:458-468.
17. Khazdair MR, Boskabady MH, Hosseini M, Rezaee R, Tsatsakis AM. The effects of Crocus sativus (saffron) and its constituents on nervous system: A review. Avicenna J Phytomed 2015;5:376-391.
18. Mehdizadeh R, Parizadeh MR, Khooei AR, Mehri S, Hosseinzadeh H. Cardioprotective effect of saffron extract and safranal in isoproterenol-induced myocardial infarction in wistar rats. Iran J Basic Med Sci 2013;16:56-63.
19. Razavi BM, Hosseinzadeh H, Movassaghi AR, Imenshahidi M, Abnous K. Protective effect of crocin on diazinon induced cardiotoxicity in rats in subchronic exposure. Chem Biol Interact 2013;203:547-555.
20. 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.
21. Riahi-Zanjani B, Balali-Mood M, Mohammadi E, Badie-Bostan H, Memar B, Karimi G. Safranal as a safe compound to mice immune system. Avicenna J Phytomed 2015;5:441-449.
22. Hosseini A, Razavi BM, Hosseinzadeh H. Saffron (Crocus sativus) petal as a new pharmacological target: a review. Iran J Basic Med Sci. 2018;21:1091-1099.
23. Hosseinzadeh H, Ziaee T, Sadeghi A. The effect of saffron, Crocus sativus stigma, extract and its constituents, safranal and crocin on sexual behaviors in normal male rats. Phytomedicine 2008;15:491-495.
24. 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.
25. Hashemi M, Hosseinzadeh H. A comprehensive review on biological activities and toxicology of crocetin. Food Chem Toxicol 2019;130:44-60.
26. Lari P, Abnous K, Imenshahidi M, Rashedinia M, Razavi M, Hosseinzadeh H. Evaluation of diazinon-induced hepatotoxicity and protective effects of crocin. Toxicol Ind Health 2015;31:367-376.
27. Hadizadeh F, Mohajeri SA, Seifi M. Extraction and purification of crocin from saffron stigmas employing a simple and efficient crystallization method. Pak J Biol Sci 2010;13:691-698.
28. Mohammadzadeh L, Hosseinzadeh H, Abnous K, Razavi BM. Neuroprotective potential of crocin against malathion-induced motor deficit and neurochemical alterations in rats. Environ Sci Pollut Res Int 2018;25:4904-4914.
29. Dorri SA, Hosseinzadeh H, Abnous K, Hasani FV, Robati RY, Razavi BM. Involvement of brain-derived neurotrophic factor (BDNF) on malathion induced depressive-like behavior in subacute exposure and protective effects of crocin. Iran J Basic Med Sci 2015;18:958-966.
30. Niehaus W, Samuelsson B. Formation of malonaldehyde from phospholipid arachidonate during microsomal lipid peroxidation. FEBS J 1968;6:126-130.
31. Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979;582:67-78.
32. Pourbakhsh H, Taghiabadi E, Abnous K, Hariri AT, Hosseini SM, Hosseinzadeh H. Effect of Nigella sativa fixed oil on ethanol toxicity in rats. Iran J Basic Med Sci 2014;17:1020-1031.
33. Hosseini SM, Taghiabadi E, Abnous K, Hariri AT, Pourbakhsh H, Hosseinzadeh H. Protective effect of thymoquinone, the active constituent of Nigella sativa fixed oil, against ethanol toxicity in rats. Iran J Basic Med Sci 2017;20:927-939.
34. Carrero RJ, Husain K. Renal oxidative stress in chronic alcohol-induced hypertension. Ethn Dis 2005;15:17-19.
35. Amrani A, Boubekri N, Lassad S, Zama D, Benayache F, Benayache S. Alcohol induced hepato cardiotoxicity and oxidative damage in rats: The protective effect of n-butanol extract of green tea (Camellia sinensis (L.) Kuntze). Cardiovasc Hematol Disord Drug Targets 2017;17:18-23.
36. Subhashree BMD, Anurupa S, Gouriprosad D. Protective role of crude extract of Amorphophallus campanulatus against ethanol-induced oxidative renal damage. Asian J Pharm Clin Res 2018;11:240-250.
37. Dinu D, Nechifor MT, Movileanu L. Ethanol‐induced alterations of the antioxidant defense system in rat kidney. J Biochem Mol Toxicol 2006;19:386-395.
38. Oyenihi OR, Afolabi BA, Oyenihi AB, Ogunmokun OJ, Oguntibeju OO. Hepato-and neuro-protective effects of watermelon juice on acute ethanol-induced oxidative stress in rats. Toxicology reports 2016;3:288-294.
39. Arslan Ö, Akdemir FNE. Antioxidant effect of trans-3 hydroxycinnamic acid against liver damage methotrexate induced in rats. J Anim Plant Sci 2018;28:1574-1578.
40. Akdemir FNE, Albayrak M, Çalik M, Bayir Y, Gülçin I. The protective effects of p-coumaric acid on acute liver and kidney damages induced by cisplatin. Biomedicines 2017;5:18-29.
41. Akdemir FNE, Gülçin I, Gürsul C, Alwasel SH, Bayir Y. Effect of p-coumaric acid against oxidative stress induced by cisplatin in brain tissue of rats. Biomedicines 2017;27:1560-1564.
42. Eki̇nci̇-Akdemi̇r FN, Yildirim S, Kandemi̇r FM, Gülçi̇n İ, Küçükler S, Sağlam YS, Yakan S. The effects of casticin and myricetin on liver damage induced by methotrexate in rats. Iran J Basic Med Sci 2018;21:1281-1288.
43. Cao YW, Jiang Y, Zhang DY, Wang M, Chen WS, Su H, et al. Protective effects of Penthorum chinense Pursh against chronic ethanol-induced liver injury in mice. J Ethnopharmacol 2015;161:92-98.
44. Akomolafe S, Oboh G, Akindahunsi A, Afolayan A. Tetracarpidium conophorum ameliorates oxidative reproductive toxicity induced by ethanol in male rats. BMC Complement Altern Med 2015;15:439-448.
45. Shanmugam K, Ramakrishna C, Mallikarjuna K, Reddy KS. Protective effect of ginger against alcohol-induced renal damage and antioxidant enzymes in male albino rats. Indian J Exp Biol 2010;48:143-149.
46. Saravanan N, Nalini N. Hemidesmus indicus protects against ethanol-induced liver toxicity. Cell Mol Biol Lett 2008;13:20-37.
47. Kamoun Z, Kamoun AS, Bougatef A, Kharrat RM, Youssfi H, Boudawara T, et al. Hepatoprotective and nephroprotective effects of sardinelle (Sardinella aurita) protein hydrolysate against ethanol-induced oxidative stress in rats. Environ Sci Pollut Res Int 2017;24:1432-1441.
48. Sivaraj A, Vinothkumar P, Sathiyaraj K, Sundaresan S, Devi K, Senthilkumar B. Hepatoprotective potential of Andrographis paniculata aqueous leaf extract on ethanol induced liver toxicity in albino rats. J Appl Pharm Sci 2011;1:204-208.
49. Lu C, Zhang F, Xu W, Wu X, Lian N, Jin H, et al. Curcumin attenuates ethanol‐induced hepatic steatosis through modulating Nrf2/FXR signaling in hepatocytes. IUBMB life 2015;67:645-658.
50. Bulle S, Reddy VD, Hebbani AV, Padmavathi P, Challa C, Puvvada PK, et al. Nephro-protective action of P. santalinus against alcohol-induced biochemical alterations and oxidative damage in rats. Biomed Pharmacother 2016;84:740-746.
51. Gutierrez-Ruiz M, Gomez LQ, Hernandez E, Bucio L, Souza V, Llorente L, et al. Cytokine response and oxidative stress produced by ethanol, acetaldehyde and endotoxin treatment in HepG2 cells. Isr Med Assoc J 2001;3:131-136.
52. Lee SH, Song YS, Jeong Y, Ko KS. Antioxidative and Anti-Inflammatory Activities of Akebia quinata Extracts in an in vitro Model of Acute Alcohol-Induced Hepatotoxicity. J Med Food 2017;20:912-922.
53. Zhen-Ming L, Wen-Yi T, Xi-Liang Z, Hui-Zi F, Zong-Hua A. Protective effects of mycelia of Antrodia camphorata and Armillariella tabescens in submerged culture against ethanol-induced hepatic toxicity in rats. J Ethnopharmacol 2007;110:160-164.
54. Das M, Basu S, Banerjee B, Sen A, Jana K, Datta G. Hepatoprotective effects of green Capsicum annum against ethanol induced oxidative stress, inflammation and apoptosis in rats. J Ethnopharmacol 2018;227:69-81.
55. Peng HC, Chen JR, Chen YL, Yang SC, Yang SS. β‐Carotene exhibits antioxidant and anti‐apoptotic properties to prevent ethanol‐induced cytotoxicity in isolated rat hepatocytes. Phytother Res 2010;24:183-189.
56. Liu Y, Zuo H, Wang Y, Tian L, Xu X, Xiong J, et al. Ethanol promotes apoptosis in rat ovarian granulosa cells via the Bcl-2 family dependent intrinsic apoptotic pathway. Cell Mol Biol 2018;64:118-125.
57. Rezaee-Khorasany AR, Razavi BM, Taghiabadi E, Tabatabaei Yazdi A, Hosseinzadeh H. Effect of saffron (stigma of Crocus sativus L.) aqueous extract on ethanol toxicity in rats: A biochemical, histopathological and molecular study. J Ethnopharmacol 2019;237:286-299.