Skip to main content

Advertisement

SpringerLink
Log in

All-trans retinoic acid mitigates methotrexate-induced liver injury in rats; relevance of retinoic acid signaling pathway

  • Original Article
  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Abstract

Methotrexate (MTX) is a widely used drug for treatment of rheumatic and autoimmune diseases as well as different types of cancer. One of the major side effects of MTX is hepatotoxicity. Retinoid receptors, including retinoid X receptor (RXR), and retinoic acid receptor (RAR) are vitamin A receptors that are highly expressed in the liver and regulate important physiological processes through regulation of different genes. In this study, we investigated the effect of MTX on RXR-α and RAR-α expression in the liver and the potential protective effects of all-trans retinoic acid (ATRA) in MTX-induced hepatotoxicity. Rats were randomly divided into five groups: The rates were treated with saline, DMSO, MTX (20 mg/kg/IP; single dose), ATRA (7.5 mg/kg/day, I.P), or MTX and ATRA. Rats were killed 24 h after the last ATRA injection. The liver tissues were dissected out, weighed, and subjected to histological, immunohistochemical, and biochemical examinations. Our results demonstrated that treatment with MTX resulted in significant decrease in reduced glutathione (GSH) content and superoxide dismutase (SOD) activity, with concomitant increase in ALT, AST, and MDA levels. In addition, MTX markedly downregulated the expression of both RXR-α and RAR-α, and changed the appearance of RXR-α to be very small speckled droplets. Treatment with ATRA significantly ameliorated MTX-induced effects on GSH, ALT, and MDA. Moreover, ATRA administration increased the expression and nuclear translocation of RXR-α in rat hepatocytes. In conclusion, our study revealed, for the first time, that retinoid receptors may play an important role in the MTX-induced hepatotoxicity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Abdel-Bakky MS, Hammad MA, Walker LA, Ashfaq MK (2010) Loss of retinoic Acid receptor (RAR) and retinoid X receptor (RXR) contribute to liver toxicity in MCT/LPS co-treated mice model. Planta Med 76:100

    Article  Google Scholar 

  • Abdel-Bakky MS, Hammad MA, Walker LA, Ashfaq MK (2011) Tissue factor dependent liver injury causes release of retinoid receptors (RXR-alpha and RAR-alpha) as lipid droplets. Biochem Biophys Res Commun 410:146–151

    Article  CAS  PubMed  Google Scholar 

  • Akbulut S, Elbe H, Eris C, Dogan Z, Toprak G, Otan E, Erdemli E, Turkoz Y (2014) Cytoprotective effects of amifostine, ascorbic acid and N-acetylcysteine against methotrexate-induced hepatotoxicity in rats. World J Gastroenterol 20:10158–10165. doi:10.3748/wjg.v20.i29.10158

    Article  PubMed Central  PubMed  Google Scholar 

  • Balmer JE, Blomhoff R (2002) Gene expression regulation by retinoic acid. J Lipid Res 43:1773–1808

    Article  CAS  PubMed  Google Scholar 

  • Beigneux AP, Moser AH, Shigenaga JK, Grunfeld C, Feingold KR (2000) The acute phase response is associated with retinoid X receptor repression in rodent liver. J Biol Chem 275:16390–16399. doi:10.1074/jbc.M000953200

    Article  CAS  PubMed  Google Scholar 

  • BEUTLER E, DURON O, KELLY BM (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–888

    CAS  PubMed  Google Scholar 

  • Bode JG, Peters-Regehr T, Gressner AM, Haussinger D (1998) De novo expression of glutamine synthetase during transformation of hepatic stellate cells into myofibroblast-like cells. Biochem J 335(Pt 3):697–700

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Dalaklioglu S, Genc GE, Aksoy NH, Akcit F, Gumuslu S (2013) Resveratrol ameliorates methotrexate-induced hepatotoxicity in rats via inhibition of lipid peroxidation. Hum Exp Toxicol 32:662–671. doi:10.1177/0960327112468178

    Article  CAS  PubMed  Google Scholar 

  • Gilani ST, Khan DA, Khan FA, Ahmed M (2012) Adverse effects of low dose methotrexate in rheumatoid arthritis patients. J Coll Phys Surg Pak 22:101–104

    Google Scholar 

  • Gudas LJ (2012) Emerging roles for retinoids in regeneration and differentiation in normal and disease states. Biochim Biophys Acta 1821:213–221. doi:10.1016/j.bbalip.2011.08.002

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Gudas LJ, Sporn MB, Roberts AB (1994) Cellular biology and biochemistry of the retinoids. The retinoids: biology, chemistry, and medicine 1994:443-520

  • Hadi NR, Al-Amran FG, Swadi A (2012) Metformin ameliorates methotrexate-induced hepatotoxicity. J Pharmacol Pharmacother 3:248–253. doi:10.4103/0976-500X.99426

    PubMed Central  CAS  PubMed  Google Scholar 

  • Jaskiewicz K, Voigt H, Blakolmer K (1996) Increased matrix proteins, collagen and transforming growth factor are early markers of hepatotoxicity in patients on long-term methotrexate therapy. J Toxicol Clin Toxicol 34:301–305

    Article  CAS  PubMed  Google Scholar 

  • Lee WM (2003) Drug-induced hepatotoxicity. N Engl J Med 349:474–485. doi:10.1056/NEJMra021844

    Article  CAS  PubMed  Google Scholar 

  • Mangelsdorf DJ, Evans RM (1995) The RXR heterodimers and orphan receptors. Cell 83:841–850

    Article  CAS  PubMed  Google Scholar 

  • Mezaki Y, Yamaguchi N, Yoshikawa K, Miura M, Imai K, Itoh H, Senoo H (2009) Insoluble, speckled cytosolic distribution of retinoic acid receptor alpha protein as a marker of hepatic stellate cell activation in vitro. J Histochem Cytochem 57:687–699. doi:10.1369/jhc.2009.953208

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Mezaki Y, Yoshikawa K, Yamaguchi N, Miura M, Imai K, Kato S, Senoo H (2007) Rat hepatic stellate cells acquire retinoid responsiveness after activation in vitro by post-transcriptional regulation of retinoic acid receptor alpha gene expression. Arch Biochem Biophys 465:370–379. doi:10.1016/j.abb.2007.06.024

    Article  CAS  PubMed  Google Scholar 

  • Mukherjee R, Davies PJ, Crombie DL, Bischoff ED, Cesario RM, Jow L, Hamann LG, Boehm MF, Mondon CE, Nadzan AM, Paterniti JR Jr, Heyman RA (1997) Sensitization of diabetic and obese mice to insulin by retinoid X receptor agonists. Nature 386:407–410. doi:10.1038/386407a0

    Article  CAS  PubMed  Google Scholar 

  • Nishikimi M, Appaji N, Yagi K (1972) The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun 46:849–854

    Article  CAS  PubMed  Google Scholar 

  • Noy N (2010) Between death and survival: retinoic acid in regulation of apoptosis. Annu Rev Nutr 30:201–217. doi:10.1146/annurev.nutr.28.061807.155509

    Article  CAS  PubMed  Google Scholar 

  • Ohata M, Lin M, Satre M, Tsukamoto H (1997) Diminished retinoic acid signaling in hepatic stellate cells in cholestatic liver fibrosis. Am J Physiol 272:G589–G596

    CAS  PubMed  Google Scholar 

  • Radaeva S, Wang L, Radaev S, Jeong WI, Park O, Gao B (2007) Retinoic acid signaling sensitizes hepatic stellate cells to NK cell killing via upregulation of NK cell activating ligand RAE1. Am J Physiol Gastrointest Liver Physiol 293:G809–G816. doi:10.1152/ajpgi.00212.2007

    Article  CAS  PubMed  Google Scholar 

  • Rao J, Zhang C, Wang P, Lu L, Zhang F (2010) All-trans retinoic acid alleviates hepatic ischemia/reperfusion injury by enhancing manganese superoxide dismutase in rats. Biol Pharm Bull 33:869–875

    Article  CAS  PubMed  Google Scholar 

  • REITMAN S, FRANKEL S (1957) A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol 28:56–63

    CAS  PubMed  Google Scholar 

  • Salbert G, Fanjul A, Piedrafita FJ, Lu XP, Kim SJ, Tran P, Pfahl M (1993) Retinoic acid receptors and retinoid X receptor-alpha down-regulate the transforming growth factor-beta 1 promoter by antagonizing AP-1 activity. Mol Endocrinol 7:1347–1356

    CAS  PubMed  Google Scholar 

  • Satoh K (1978) Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 90:37–43

    Article  CAS  PubMed  Google Scholar 

  • Sener G, Eksioglu-Demiralp E, Cetiner M, Ercan F, Sirvanci S, Gedik N, Yegen BC (2006a) L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death. Cell Biol Toxicol 22:47–60. doi:10.1007/s10565-006-0025-0

    Article  CAS  PubMed  Google Scholar 

  • Sener G, Eksioglu-Demiralp E, Cetiner M, Ercan F, Yegen BC (2006b) Beta-glucan ameliorates methotrexate-induced oxidative organ injury via its antioxidant and immunomodulatory effects. Eur J Pharmacol 542:170–178. doi:10.1016/j.ejphar.2006.02.056

    Article  CAS  PubMed  Google Scholar 

  • Ulven SM, Natarajan V, Holven KB, Lovdal T, Berg T, Blomhoff R (1998) Expression of retinoic acid receptor and retinoid X receptor subtypes in rat liver cells: implications for retinoid signalling in parenchymal, endothelial, Kupffer and stellate cells. Eur J Cell Biol 77:111–116

    Article  CAS  PubMed  Google Scholar 

  • Uraz S, Tahan V, Aygun C, Eren F, Unluguzel G, Yuksel M, Senturk O, Avsar E, Haklar G, Celikel C, Hulagu S, Tozun N (2008) Role of ursodeoxycholic acid in prevention of methotrexate-induced liver toxicity. Dig Dis Sci 53:1071–1077. doi:10.1007/s10620-007-9949-3

    Article  CAS  PubMed  Google Scholar 

  • Vogel S, Piantedosi R, Frank J, Lalazar A, Rockey DC, Friedman SL, Blaner WS (2000) An immortalized rat liver stellate cell line (HSC-T6): a new cell model for the study of retinoid metabolism in vitro. J Lipid Res 41:882–893

    CAS  PubMed  Google Scholar 

  • Vonen B, Morland J (1984) Isolated rat hepatocytes in suspension: potential hepatotoxic effects of six different drugs. Arch Toxicol 56:33–37

    Article  CAS  PubMed  Google Scholar 

  • Wang H, Dan Z, Jiang H (2008) Effect of all-trans retinoic acid on liver fibrosis induced by common bile duct ligation in rats. J Huazhong Univ Sci Technol Med Sci 28:553–557. doi:10.1007/s11596-008-0514-x

    Article  PubMed  Google Scholar 

  • Wu JJ, Schiff KR (2004) Sarcoidosis. Am Fam Physician 70:312–322

    PubMed  Google Scholar 

  • Xiao JH, Durand B, Chambon P, Voorhees JJ (1995) Endogenous retinoic acid receptor (RAR)-retinoid X receptor (RXR) heterodimers are the major functional forms regulating retinoid-responsive elements in adult human keratinocytes. Binding of ligands to RAR only is sufficient for RAR-RXR heterodimers to confer ligand-dependent activation of hRAR beta 2/RARE (DR5). J Biol Chem 270:3001–3011

    Article  CAS  PubMed  Google Scholar 

Download references

Conflict of interest

The authors declare that they have no competing interest.

Author information

Authors and Affiliations

  1. Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt

    Mohamed G. Ewees, Tamer M. Abdelghany, Abdel-Aziz H. Abdel-Aziz & Mohamed S. Abdel-Bakky

  2. College of Pharmacy, Aljouf University, Aljouf, 2014, Kingdom of Saudi Arabia

    Mohamed S. Abdel-Bakky

Authors
  1. Mohamed G. Ewees
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tamer M. Abdelghany
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Abdel-Aziz H. Abdel-Aziz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mohamed S. Abdel-Bakky
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tamer M. Abdelghany.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ewees, M.G., Abdelghany, T.M., Abdel-Aziz, AA.H. et al. All-trans retinoic acid mitigates methotrexate-induced liver injury in rats; relevance of retinoic acid signaling pathway. Naunyn-Schmiedeberg's Arch Pharmacol 388, 931–938 (2015). https://doi.org/10.1007/s00210-015-1130-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00210-015-1130-5

Keywords

Search

Navigation