Protective effects of crocin against hepatic damages in D-galactose aging model in rats

Document Type : Research article


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

2 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Pharmaceutical 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, I. R. Iran

4 Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran. Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.


Aging is a progressive process which is associated with liver dysfunction and it is due to oxidative stress, inflammation, and cell apoptosis. Long-term D-galactose (D-gal) administration is able to develop an aging model in animals. Crocin as a major active ingredient in saffron has shown anti-inflammatory and hepatoprotective effects via its antioxidant capacity. Thus, the aim of the present study was the assessment of crocin effects on hepatic and metabolic disorders induced by D-gal in rats. Aging model was induced in rats by 56-day administration of D-gal (400 mg/kg/day subcutaneously). Protective effects of different doses of crocin (7.5, 15 and 30 mg/kg/day) in concomitant with D-gal administration were evaluated. Malondialdehyde (MDA) and reduced glutathione (GSH) amounts were measured by means of their reaction, respectively, with thiobarbituric acid and 5,5′-Dithiobis (2-nitrobenzoic acid) (DTNB) in a specific condition. Cyclooxygenase-2 (COX-2), β-galactosidase, induced nitric oxide synthase (iNOS), and carboxymethyllysine (CML) levels were determined by western blotting method. Additionally, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were measured in serum. D-gal administration significantly elevated ALT, AST, ALP levels, which were markedly inhibited by crocin administration. Crocin suppressed the overgeneration of lipid peroxidation products such as MDA. iNOS was elevated by D-gal administration and was returned to the normal extent by crocin. Therefore, Crocin as a powerful antioxidant and radical scavenger, totally exhibited hepatoprotective effects against D-gal-induced toxicity in rats

Graphical Abstract

Protective effects of crocin against hepatic damages in D-galactose aging model in rats


  1. References

    1. Khansari N, Shakiba Y and Mahmoudi M. Chronic inflammation and oxidative stress as a major cause of age-related diseases and cancer. Recent Pat. Inflamm. Allergy Drug Discov. (2009) 3: 73-80.
    2. Matjusaitis M, Chin G, Sarnoski EA and Stolzing A. Biomarkers to identify and isolate senescent cells. Ageing Res. Rev. (2016) 29: 1-12.
    3. Afanas'ev I. Superoxide and nitric oxide in senescence and aging. Front. Biosci. (Landmark Ed). (2009) 14: 3899-912.
    4. Patruno A, Speranza L, Tete S, Mastrangelo F, De Lutiis MA, Pesce M, Franceschelli S, Vinciguerra I, Felaco M and Grilli A. iNOS activity in the aged rat liver tissue. J. Biol. Regul. Homeost. Agents (2007) 21: 89-95.
    5. Buttke TM and Sandstrom PA. Oxidative stress as a mediator of apoptosis. Immunol. Today (1994) 15: 7-10.
    6. Gill R, Tsung A and Billiar T. Linking oxidative stress to inflammation: Toll-like receptors. Free Radic Biol. Med. (2010) 48: 1121-32.
    7. Busch CJ and Binder CJ. Malondialdehyde epitopes as mediators of sterile inflammation. Biochim. Biophys. Acta (2016).
    8. Kizer JR, Benkeser D, Arnold AM, Ix JH, Mukamal KJ, Djousse L, Tracy RP, Siscovick DS, Psaty BM and Zieman SJ. Advanced glycation/glycoxidation endproduct carboxymethyl-lysine and incidence of coronary heart disease and stroke in older adults. Atherosclerosis (2014) 235: 116-21.
    9. Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D and Hwang ES. Senescence-associated beta-galactosidase is lysosomal beta-galactosidase. Aging Cell. (2006) 5: 187-95.
    10. Baynes JW. The role of AGEs in aging: causation or correlation. Exp. Gerontol. (2001) 36: 1527-37.
    11. Kalaz EB, Coban J, Aydin AF, Dogan-Ekici I, Dogru-Abbasoglu S, Oztezcan S and Uysal M. Carnosine and taurine treatments decreased oxidative stress and tissue damage induced by D-galactose in rat liver. J. Physiol. Biochem. (2014) 70: 15-25.
    12. An F, Yang G, Tian J and Wang S. Antioxidant effects of the orientin and vitexin in Trollius chinensis Bunge in D-galactose-aged mice. Neural Regen. Res. (2012) 7: 2565-75.
    13. Coban J, Betul-Kalaz E, Kucukgergin C, Aydin AF, Dogan-Ekici I, Dogru-Abbasoglu S and Uysal M. Blueberry treatment attenuates D-galactose-induced oxidative stress and tissue damage in rat liver. Geriatr. Gerontol. Int. (2014) 14: 490-7.
    14. Feng Y, Yu YH, Wang ST, Ren J, Camer D, Hua YZ, Zhang Q, Huang J, Xue DL, Zhang XF, Huang XF and Liu Y. Chlorogenic acid protects D-galactose-induced liver and kidney injury via antioxidation and anti-inflammation effects in mice. Pharm. Biol. (2016) 54: 1027-34.
    15. Ruan Q, Liu F, Gao Z, Kong D, Hu X, Shi D, Bao Z and Yu Z. The anti-inflamm-aging and hepatoprotective effects of huperzine A in D-galactose-treated rats. Mech. Ageing Dev. (2013) 134: 89-97.
    16. Song X, Bao M, Li D and Li YM. Advanced glycation in D-galactose induced mouse aging model. Mech. Ageing Dev. (1999) 108: 239-51.
    17. Oh YS, Seo EH, Lee YS, Cho SC, Jung HS, Park SC and Jun HS. Increase of Calcium Sensing Receptor Expression Is Related to Compensatory Insulin Secretion during Aging in Mice. PLoS One (2016) 11: e0159689.
    18. Salmon AB. Beyond Diabetes: Does Obesity-Induced Oxidative Stress Drive the Aging Process? Antioxidants (Basel). (2016) 5.
    19. Mollazadeh H, Emami SA and Hosseinzadeh H. Razi's Al-Hawi and saffron (Crocus sativus): a review. Iran. J. Basic Med. Sci. (2015) 18: 1153-66.
    20. Khorasany AR and Hosseinzadeh H. Therapeutic effects of saffron (Crocus sativus L.) in digestive disorders: a review. Iran. J. Basic Med. Sci. (2016) 19: 455-69.
    21. Boskabady MH and Farkhondeh T. Antiinflammatory, Antioxidant, and Immunomodulatory Effects of Crocus sativus L. and its Main Constituents. Phytother. Res. (2016) 30: 1072-94.
    22. Assimopoulou AN, Sinakos Z and Papageorgiou VP. Radical scavenging activity of Crocus sativus L. extract and its bioactive constituents. Phytother. Res. (2005) 19: 997-1000.
    23. Alavizadeh SH and Hosseinzadeh H. Bioactivity assessment and toxicity of crocin: a comprehensive review. Food Chem. Toxicol. (2014) 64: 65-80.
    24. Boussabbeh M, Ben Salem I, Belguesmi F, Neffati F, Najjar MF, Abid-Essefi S and Bacha H. Crocin protects the liver and kidney from patulin-induced apoptosis in-vivo. Environ. Sci. Pollut. Res. Int. (2016) 23: 9799-808.
    25. Hosseinzadeh H, Abootorabi A and Sadeghnia HR. Protective effect of Crocus sativus stigma extract and crocin (trans-crocin 4) on methyl methanesulfonate-induced DNA damage in mice organs. DNA Cell Biol. (2008) 27: 657-64.
    26. Lv B, Huo F, Zhu Z, Xu Z, Dang X, Chen T, Zhang T and Yang X. Crocin Upregulates CX3CR1 Expression by Suppressing NF-kappaB/YY1 Signaling and Inhibiting Lipopolysaccharide-Induced Microglial Activation. Neurochem. Res. (2016) 41: 1949-57.
    27. Lari P, Abnous K, Imenshahidi M, Rashedinia M, Razavi M and Hosseinzadeh H. Evaluation of diazinon-induced hepatotoxicity and protective effects of crocin. Toxicol. Ind. Health (2013) 31: 367-76.
    28. Razavi BM and Hosseinzadeh H. Saffron: a promising natural medicine in the treatment of metabolic syndrome. J. Sci. Food Agric. (2017) 97: 1679-85.
    29. Shahroudi MJ, Mehri S and Hosseinzadeh H. Anti-Aging Effect of Nigella Sativa Fixed Oil on D-Galactose-Induced Aging in Mice. J. Pharmacopuncture (2017) 20: 29-35.
    30. Mohammadi E MS, Badie Bostan H and Hosseinzadeh H. Protective effect of crocin against d-galactose-induced aging in mice. Avicenna J. Phytomed. (2018) 8:14-23.
    31. Hadizadeh F, Mohajeri SA and Seifi M. Extraction and purification of crocin from saffron stigmas employing a simple and efficient crystallization method. Pak. J. Biol. Sci. (2010) 13: 691-8.
    32. Bandegi AR, Rashidy-Pour A, Vafaei AA and Ghadrdoost B. Protective Effects of Crocus Sativus L. Extract and Crocin against Chronic-Stress Induced Oxidative Damage of Brain, Liver and Kidneys in Rats. Adv. Pharm. Bull. (2014) 4: 493-9.
    33. Salahshoor MR, Khashiadeh M, Roshankhah S, Kakabaraei S and Jalili C. Protective effect of crocin on liver toxicity induced by morphine. Res. Pharm. Sci. (2016) 11: 120-9.
    34. Mihara M and Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal. Biochem. (1978) 86: 271-8.
    35. Moron MS, Depierre JW and Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim. Biophys. Acta (1979) 582: 67-78.
    36. Huang CC, Chiang WD, Huang WC, Huang CY, Hsu MC and Lin WT. Hepatoprotective Effects of Swimming Exercise against D-Galactose-Induced Senescence Rat Model. Evid. Based Complement Alternat. Med. (2013) 2013: 275431.
    37. Hadzi-Petrushev N, Stojkovski V, Mitrov D and Mladenov M. D-galactose induced changes in enzymatic antioxidant status in rats of different ages. Physiol. Res. (2015) 64: 61-70.
    38. Samarghandian S, Azimi-Nezhad M, Borji A and Farkhondeh T. Effect of crocin on aged rat kidney through inhibition of oxidative stress and proinflammatory state. Phytother. Res. (2016): 1345-53.
    39. Kasapoglu M and Ozben T. Alterations of antioxidant enzymes and oxidative stress markers in aging. Exp. Gerontol. (2001) 36: 209-20.
    40. Bathina S, Srinivas N and Das UN. Streptozotocin produces oxidative stress, inflammation and decreases BDNF concentrations to induce apoptosis of RIN5F cells and type 2 diabetes mellitus in Wistar rats. Biochem Biophys. Res. Commun (2017).
    41. Shirali S, Zahra Bathaie S and Nakhjavani M. Effect of crocin on the insulin resistance and lipid profile of streptozotocin-induced diabetic rats. Phytother. Res. (2013) 27: 1042-47.
    42. Boyle DL, Blunt DS and Takemoto LJ. Confocal microscopy of cataracts from animal model systems: relevance to human nuclear cataract. Exp. Eye. Res. (1997) 64: 565-72.
    43. Lackner PA, Rodriguez L, Sato S, Lizak MJ, Wyman M and Kador PF. Age-dependent lens changes in galactose-fed dogs. Exp. Eye. Res. (1997) 64: 431-6.
    44. Engerman RL and Kern TS. Experimental Galactosemia Produces Diabetic-like Retinopathy. Diabetes (1984) 33: 97.
    45. Afanas'ev IB. Free radical mechanisms of aging processes under physiological conditions. Biogerontology (2005) 6: 283-90.
    46. Fan SH, Zhang ZF, Zheng YL, Lu J, Wu DM, Shan Q, Hu B and Wang YY. Troxerutin protects the mouse kidney from d-galactose-caused injury through anti-inflammation and anti-oxidation. Int. Immunopharmacol. (2009) 9: 91-6.
    47. Hsieh HM, Wu WM and Hu ML. Genistein attenuates D-galactose-induced oxidative damage through decreased reactive oxygen species and NF-kappaB binding activity in neuronal PC12 cells. Life Sci. (2011) 88: 82-8.
    48. Kirkby NS, Chan MV, Zaiss AK, Garcia-Vaz E, Jiao J, Berglund LM, Verdu EF, Ahmetaj-Shala B, Wallace JL, Herschman HR, Gomez MF and Mitchell JA. Systematic study of constitutive cyclooxygenase-2 expression: Role of NF-kappaB and NFAT transcriptional pathways. Proc. Natl. Acad. Sci. U S A. (2016) 113: 434-9.
    49. Poligone B and Baldwin AS. Positive and negative regulation of NF-kappaB by COX-2: roles of different prostaglandins. J. Biol. Chem. (2001) 276: 38658-64.
    50. Li C, Mo Z, Xie J, Xu L, Tan L, Luo D, Chen H, Yang H, Li Y, Su Z and Su Z. Chongcao-Shencha Attenuates Liver and Kidney Injury through Attenuating Oxidative Stress and Inflammatory Response in D-Galactose-Treated Mice. Evid Based Complement Alternat. Med. (2016) 2016: 3878740.
    51. Ullah F, Ali T, Ullah N and Kim MO. Caffeine prevents d-galactose-induced cognitive deficits, oxidative stress, neuroinflammation and neurodegeneration in the adult rat brain. Neurochem. Int. (2015) 90: 114-24.
    52. Badawi AF, Liu Y, Eldeen MB, Morrow W, Razak ZR, Maradeo M and Badr MZ. Age-associated changes in the expression pattern of cyclooxygenase-2 and related apoptotic markers in the cancer susceptible region of rat prostate. Carcinogenesis (2005) 26: 1650.
    53. Yan D, Liu H-L, Yu Z-J, Huang Y-H, Gao D, Hao H, Liao S-S, Xu F-Y and Zhou X-Y. BML-111 Protected LPS/D-GalN-Induced Acute Liver Injury in Rats. Int. J. Mol. Sci. (2016) 17.