• Heba Abdulmohsin Department of Pharmacology, College of Medicine, Al-Nahrain University, Baghdad, Iraq.
  • Ahmed Abu Raghif Department of Pharmacology, College of Medicine, Al-Nahrain University, Baghdad, Iraq.
  • Mohammed Jabbar Manna Department of , College of Dentistry, Al-Mustansiriya University, Baghdad, Iraq.


Objective: The aim of this study was to investigate antioxidant and hepatoprotective properties of Iraqi Echinops heterophyllus aqueous crude extract and its flavonoid fraction against methotrexate (MTX)-induced hepatotoxicity in rabbits.

Methods: MTX-induced hepatotoxicity by administration of 20 mg/kg MTX intraperitoneally for 3 successive days was used as animal model, and animals were arrayed in four groups with eight animals in each group: Group 1 was the healthy control, Group 2 - the negative control receiving MTX only, Group 3 received MTX+crude extract of E. heterophyllus, and Group 4 administered MTX+flavonoid fraction of E. heterophyllus. The study duration was 10 days; at day 11, animals were sacrificed, and the blood samples were obtained for the measurement of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total bilirubin, total protein, and albumin as well as ELISA assay of the oxidative stress markers such as glutathione (GSH) and malondialdehyde (MDA). The liver was dissected and processed for histopathological investigation and scoring. Statistical analysis was performed to investigate the significance of each result.

Results: The study results revealed severe liver damage due to MTX administration in the negative control (induced-non treated) group in comparison with healthy group, also there was significant hepatoprotective effect after administration of the crude extract of E. heterophyllus, and flavonoid fraction from E. heterophyllus shown after biochemical liver function tests and anti-oxidant properties demonstrated by the measurement of oxidative stress markers MDA and GSH. The crude extract of E. heterophyllus shown superior hepatoprotective and antioxidant effect. Histopathological scoring showed a remarkable decrease in the scores of the treatment groups in comparison with the high score in the MTX only treated group.

Conclusions: MTX administered in a dose of 20 mg/kg for 3 successive days causes marked liver injury, while treatment with the crude extract and flavonoid fraction of E. heterophyllus significantly ameliorates MTX-induced liver damage, although the crude extract of E. heterophyllus seems to have the most hepatoprotective properties.

Keywords: Echinops heterophyllus, Flavonoids, Methotrexate, Hepatotoxicity, Antioxidant, Hepatoprotective.


1. Rider BJ. Methotrexate. Ref Modul Biomed Sci 2007;1:1-5.
2. Vohra R, Vohra SS, Grock A, Mason J. Working through the paradox of methotrexate toxicity. Ann Emerg Med 2018;72:129-32.
3. Famurewa AC, Ufebe OG, Egedigwe CA, Nwankwo OE, Obaje GS. Virgin coconut oil supplementation attenuates acute chemotherapy hepatotoxicity induced by anticancer drug methotrexate via inhibition of oxidative stress in rats. Biomed Pharmacother 2017;87:437-42.
4. Abdulrasool AA, Fahmi ZM, Khadeem EJ. A relative assess on wound healing and anti scar activity of crude echinops heterophyllus extract and some of its bioactive fractions. Int J Pharm Pharm Sci 2013;5:468-75.
5. Fahmi ZM, Khadeem EJ, Hasan HF, Luaibi OK. The Antibacterial effect of phytosterols isolated from echinops heterophyllus in comparison with mebo ® and standard antimicrobial agents. Am J Polit Sci 2014;14:6-8.
6. Famurewa AC, Folawiyo AM, Epete MA, Igwe EC, Okike PI, Maduagwuna EK, et al. Abrogation of hepatic damage induced by anticancer drug methotrexate by zobo (Hibiscus sabdariffa extract) supplementation via targeting oxidative hepatotoxicity in rats. J Diet Suppl 2018;???:1-3.
7. Abdulrazzaq MH, Khadeem EJ, Muhammadi SS. Hepatoprotective effect of Echinops tenuisectus (Compositae) on CCl 4 induced hepatic damage in rats. Iraqi J Pharm Sci 2008;17:16-24.
8. Theise ND. Liver biopsy assessment in chronic viral hepatitis: A personal, practical approach. Mod Pathol 2007;20 Suppl 1:S3-14.
9. Wood GS, Wu J. Methotrexate and pralatrexate. Dermatol Clin 2015;33:747-55.
10. Ali N, Rashid S, Nafees S, Hasan SK, Shahid A, Majed F, et al. Protective effect of chlorogenic acid against methotrexate induced oxidative stress, inflammation and apoptosis in rat liver: An experimental approach. Chem Biol Interact 2017;272:80-91.
11. Mehrzadi S, Fatemi I, Esmaeilizadeh M, Ghaznavi H, Kalantar H, Goudarzi M, et al. Hepatoprotective effect of berberine against methotrexate induced liver toxicity in rats. Biomed Pharmacother 2018;97:233-9.
12. Khalifa MM, Bakr AG, Osman AT. Protective effects of phloridzin against methotrexate-induced liver toxicity in rats. Biomed Pharmacother 2017;95:529-35.
13. Pattanayak S, Nayak SS, Panda DP, Dinda SC. Hepatoprotective activity of crude flavonoids extract of Cajanus scarabaeoides (L) in paracetamol intoxicated albino rats. Asian J Pharm Biol Res 2011;1:22-7.
14. Mahmoud AM, Hussein OE, Hozayen WG, Abd El-Twab SM. Methotrexate hepatotoxicity is associated with oxidative stress, and down-regulation of PPARγ and nrf2: Protective effect of 18β-glycyrrhetinic acid. Chem Biol Interact 2017;270:59-72.
15. Kumari S. Methotrexate induced hepatotoxicity and its management. Int J Sci Res 2013;14:2319-7064.
16. Fontana RJ. Reviews in basic and clinical gastroenterology. Gastroenterology 2014;146:914-28.
17. Ali F, Khan KM, Salar U, Taha M, Hadiani N, Wadood A, et al. ???. Eur J Med Chem 2017;???:???.
18. Arul DJ, Das S, Jayan N, Asha DS. Protective activity of asparagus racemosus in methotrexate-induced liver toxicity in wistar rats. Asian J Pharm Clin Res 2018;11:9-12.
19. Turgut G, Enli Y, Kaptanoǧlu B, Turgut S, Genç O. Changes in the levels of MDA and GSH in mice serum, liver and spleen after aluminum administration. East J Med 2006;11:7-12.
20. Rath D, Prusty S, Sahu PK. Hepatoprotective effects of cassia tora whole plant. Int J Pharm Technol 2016;3:2798-806.
21. Pınar N, Kaplan M, Özgür T, Özcan O. Ameliorating effects of tempol on methotrexate-induced liver injury in rats. Biomed Pharmacother 2018;102:758-64.
22. Baghel SS, Mavai Y, Baghel RS, Sikarwar I, Shrivastava N. Evaluation of hepatoprotective efficacy of rhizomes Curcuma caesia in paracetamol induced hepatotoxicity in rats. Int J Pharm Pharm Sci 2013;5:249-55.
23. Cuce G, Çetinkaya S, Koc T, Esen HH, Limandal C, Balcı T, et al. Chemoprotective effect of vitamin E in cyclophosphamide-induced hepatotoxicity in rats. Chem Biol Interact 2015;232:7-11.
24. Hafez HM, Ibrahim MA, Ibrahim SA, Amin EF, Goma W, Abdelrahman AM, et al. Potential protective effect of etanercept and aminoguanidine in methotrexate-induced hepatotoxicity and nephrotoxicity in rats. Eur J Pharmacol 2015;768:1-2.
25. Khafaga AF, El-Sayed YS. Spirulina ameliorates methotrexate hepatotoxicity via antioxidant, immune stimulation, and proinflammatory cytokines and apoptotic proteins modulation. Life Sci 2018;196:9-17.
26. Hasan HT, Kadhim EJ. Phytochemical investigation of leaves and seeds of Corchorus olitorius l. Cultivated in Iraq. Asian J Pharm Clin Res 2018;11:3-12.
27. García ER, Gutierrez EA, de Melo FCSA, Novaes RD, Gonçalves RV. Flavonoids effects on hepatocellular carcinoma in murine models: A systematic review. Evid Based Complement Alternat Med 2018;2018:6328970.
28. Verri WA, Vicentini FT, Baracat MM, Georgetti SR, Cardoso RD, Cunha TM, et al. Flavonoids as anti-inflammatory and analgesic drugs: Mechanisms of action and perspectives in the development of pharmaceutical forms. Bioact Nat Prod 2012;36:297-330.
29. Xiang C, Teng Y, Yao C, Li X, Cao M, Li X, et al. Antioxidant properties of fl avonoid derivatives and their hepatoprotective effects on CCl 4 induced acute liver injury in mice. R Soc Chem 2018;8:15366-71.
30. Khan H, Jawad M, Kamal MA, Baldi A, Xiao J, Nabavi SM, et al. Evidence and prospective of plant derived flavonoids as antiplatelet agents: Strong candidates to be drugs of future. Food Chem Toxicol 2018;119:355-67.
31. Wu Y, Wang F, Zheng Q, Lu L, Yao H, Zhou C, et al. Hepatoprotective effect of total flavonoids from Laggera alata against carbon tetrachloride-induced injury in primary cultured neonatal rat hepatocytes and in rats with hepatic damage. J Biomed Sci 2006;13:569-78.
32. Abdulrazzaq MH, Khadeem EJ, Muhammadi SS. Hepatoprotective activity of bioherbal ethanolic extract on CCl 4 induced hepatic damage in rats. Iraqi J Pharm Sci 2008;17:61-5.
33. Gupta A, Sheth NR, Pandey S, Yadav JS, Joshi SV. Screening of flavonoids rich fractions of three Indian medicinal plants used for the management of liver diseases. Braz J Pharmacogn 2015;25:485-90.
34. Stevenson R. Instrumentation. J Chromatogr Libr 2004;69:469-518.
46 Views | 88 Downloads
How to Cite
Abdulmohsin, H., A. Abu Raghif, and M. J. Manna. “THE PROTECTIVE EFFECTS OF ECHINOPS HETEROPHYLLUS AQUEOUS EXTRACT AGAINST METHOTREXATE-INDUCED HEPATOTOXICITY IN RABBITS”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 12, no. 1, Jan. 2019, pp. 384-90, doi:10.22159/ajpcr.2019.v12i1.30213.
Original Article(s)