DARU Journal of Pharmaceutical Sciences

, Volume 27, Issue 2, pp 721–733 | Cite as

Ellagic acid reduces methotrexate-induced apoptosis and mitochondrial dysfunction via up-regulating Nrf2 expression and inhibiting the IĸBα/NFĸB in rats

  • Reihaneh Ebrahimi
  • Mohammad Reza Sepand
  • Seyed Afshin Seyednejad
  • Ameneh Omidi
  • Mostafa Akbariani
  • Maryam Gholami
  • Omid SabzevariEmail author
Research article



The clinical application of methotrexate (MTX), an efficacious cytotoxic drug, is restricted due to its associated liver toxicity. Ellagic acid (EA), a natural polyphenol, possesses hepatoprotective, antioxidant and anti-inflammatory properties.


The present study seeks to address the hepatoprotective effects of Ellagic acid (EA) against MTX-mediated oxidative stress (OS) and widen our current knowledge of the underlying molecular mechanisms of MTX toxicity.


Wistar rats were orally given EA (5 mg/kg and 10 mg/kg) for 10 successive days and at the end of the third day they were administered a single dose of MTX (20 mg/kg i.p).


After performing biochemical analysis, liver enzymes and malondialdehyde were significantly higher in the MTX group, indicating hepatic oxidative damage. MTX-induced OS was further confirmed with observation of events such as reactive oxygen species (ROS) overproduction, mitochondrial outer membrane potential decrease, mitochondrial swelling, cytochrome c release and caspase-3/9 increase, resulting in apoptosis. Furthermore, overexpression of pro-inflammatory factors such as nuclear factor kappa B (NF-ĸB) and interleukin 6 (IL-6) indicated the MTX-induced inflammation in MTX-treated group. Interestingly, EA was able to significantly prevent OS, mitochondrial dysfunction, apoptosis and inflammation induced by MTX. Also, EA-treated rats demonstrated significant upregulation of both nuclear factor erythroid 2-related factor 2 (Nrf2) and hemoxygenase-1 (HO-1), which were considerably downregulated in MTX-treated rats.


EA protects rats against MTX-induced apoptosis and mitochondrial dysfunction via up-Regulating Nrf2 and HO-1 expression and inhibiting the NF-κB signaling pathway. Therefore, EA may protect patients against MTX-induced hepatotoxicity and encourage its clinical application.

Graphical abstract

Beneficial effect of Ellagic acid (EA) on Methotrexate (MTX)-induced liver injury: molecular mechanism.


Ellagic acid Mitochondrial dysfunction Oxidative stress Inflammation Apoptosis 



This work was supported by Vice Chancellor for Research, Tehran University of Medical Sciences, Tehran, Iran under Grant (96-03-169-36364).

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interest regarding the publication of this article.


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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Toxicology and Poisoning Research Centre, Department of Toxicology and Pharmacology, International Campus, Faculty of PharmacyTehran University of Medical SciencesTehranIran
  2. 2.Department of Anatomical Sciences, Faculty of Medical SciencesTarbiat Modares UniversityTehranIran

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