Ameliorative Effect of Propolis Extract on Hepatotoxicity Induced by Methotrexate in Mice


  • Gehan Moustafa Badr - Physiology section, Physiology, Zoology Department, Faculty of Science, Ain Shams University, Abbassia, 11566, Cairo, Egypt - Biological Sciences Department, Faculty of Science, King Faisal University, Saudi Arabia.


Propolis, Methotrexate, liver, antioxidants, cytokines


Propolis, rich in natural antioxidant flavonoids. It is produced by honey bee (Apis melifera ligutica). Our study aimed to evaluate the ameliorative effects of Propolis water extract on liver toxicity induced by methotrexate. The study used thirty-five male albino mice randomly divided into five groups. The first is control (C); the second, Methotrexate-induced hepatotoxicity (MTX, single dose 20 mg/kg IP); Third, Propolis water extract (PWE, 100 mg/kg PO for 7 days) and both fourth and fifth groups are Proplis Before/After Methotrexate, PBM and PAM; respectively). Serum aspartate aminotransferase (ASTA), alanine aminotransferase (ALTA), alkaline phosphatase (ALP), γ-glutamyle transferase (γ-GT), C-reactive protein (CRP) and hepatic histopathology estimated the protective effect of Propolis against the liver injury induced by MTX. Catalase (CAT) and superoxide dismutase (SOD) evaluated the antioxidant activities in liver tissue. In additions, the immune efficacy of Propolis were determined by liver interleukins levels (IL4, IL10, IL1 and IL6). The results revealed that Propolis treatment before injection of Methotrexate have significant hepatoprotective, antioxidant and enhancing immune liver response compared to PAM groups. The study, assumes in line with the fact that Propolis is a natural, low cost and nontoxic product possesses ameliorative activities against hepatic alterations induced by MTX.


• AebiH E. 1983. Catalase in: method of enzymatic analysis. Vol.3,(ed) H.V. Bergmeyer Berlin: Verlag Chemi.273-286.

• AkyolS, GinisZ, Armutcu F, OzturkG, YigitogluMR., Akyol O. 2012. The potential usage of caffeic acid phenethyl ester (CAPE) against chemotherapy-induced and radiotherapy-induced toxicity. Cell BiochemFunct. 30:438–43.

• Akyol S, Gulec M A, ErdemliHK,Akyol O. 2016.Can propolis and caffeic acid phenethyl ester be promising agents against cyclophosphamide toxicity? J IntercultEthnopharmacol. 5(1): 105–107. doi: 10.5455/jice.20160127024542.

• Ali N, Rashid S, Nafees S, Hasan S K, Sultana S. 2014. Beneficial effects of Chrysin

• against Methotrexate-induced hepatotoxicity via attenuation of oxidativestress and apoptosis. Mol Cell Biochem. 385(1–2):215–23.

• Arenaa U, Stasi C, Mannonib A, Benuccic M, Maddali-Bongid S, Cammellia D, Assarata A, Marraa F, PinzaniaM. 2012. Liver stiffness correlates with methotrexate cumulative dose in patients with rheumatoid arthritis. Digestive and Liver Disease. 44: 149–153.

• Aslaner A, Çakır T, Çelik B, Doğan U, Akyüz C, Baştürk A, Polat C, Gündüz U, Mayir B, Şehirli Ö. 2015. The protective effect of intraperitoneal medical ozone preconditioning and treatment on hepatotoxicity induced by methotrexate. Int J ClinExp Med. 8(8):13303-13309.

• Babatunde R I, Abdulbasit A, Oladayo M I, Olasile O I, Olamide F R, Gbolahan B W. 2015. Hepatoprotective and Pancreatoprotective Properties of the Ethanolic Extract of Nigerian Propolis. J IntercultEthnopharmacol. 4(2):102-8. doi: 10.5455/jice.20150202023615.

• Bancroft J D, Gamble M. 2008. Theory and practice of histological techniques. Fifth ed. Churchill Livingstone. London. pp: 109-136.

• Banji D, Pinnapareddy J, Banji O J F, Kumar R A, Reddy N K. 2011. Evaluation of the concomitant use of methotrexate and curcumin on Freund's complete adjuvant-induced arthritis and hematological indices in rats. Ind. J. Pharmacol. 43:548–550.

• Barker J,Horn E J, Lebwohl M, Warren R B, NastA, Rosenberg W, Smith C. 2011. Assessment and management of methotrexate hepatotoxicity in psoriasis patients: Report from a consensus conference to evaluate current practice and identify key questions toward optimizing methotrexate use in the clinic International Psoriasis. J. Eur. Acad. Dermatol. Venereol. 25: 758–764.

• Belfield A, Goldberg D M. 1971. Colorimetric determination of alkaline phosphatase activity. Enzymes. 12: 561-566.

• Campos JF, Dos SantosUP, da Rocha Pdos S, DamiãoMJ, BalestieriJB, CardosoCA, Paredes-GameroEJ, EstevinhoLM, de Picoli-SouzaK, Dos SantosEL 2015.Antimicrobial, Antioxidant, Anti-Inflammatory, and Cytotoxic Activities of Propolis from the Stingless BeeTetragoniscafiebrigi (Jataí).Evid Based Complement Alternat Med. 2015;2015:296186. doi: 10.1155/2015/296186. Epub 2015 Jun 22.

• Chladek J, Martinkova J, Sispera L. 1997. An in vitro study on methotrexate hydroxylation in rat and human liver. Physiol Res. 46: 371-9.

• De S, Sen T, Chatterjee M. 2015. Reduction of oxidative stress by an ethanolic extract of leaves of Piper betle (Paan) Linn. Decreased methotrexate-induced toxicity.Mol Cell Biochem. 409(1-2):191-197. doi: 10.1007/s11010-015-2524-x.

• Duman D G, Kumral Z N Ö, Ercan, F, Deniz M, Can G, ÇağlayanYeğen, B. 2013. Saccharomyces boulardii ameliorates clarithromycin-and methotrexate inducedintestinal and hepatic injury in rats. Br J Nutr.110(03):493–9.

• El Sohaimy S A., Masry S H D. 2014.Phenolic Content, Antioxidant and Antimicrobial Activities of Egyptian and Chinese Propolis. American-Eurasian J. Agric. & Environ. Sci. 14 (10): 1116-1124.

• El-Aidy W K, Ebeid A A, Sallam E M,Muhammad I E.,Abbas A T, Kamal M A, Sohrab S S. 2015. Evaluation of propolis, honey, and royal jelly in amelioration of peripheral blood leukocytes and lung inflammation in mouse conalbumin-induced asthma model. Saudi J Biol Sci. 22(6): 780–788.

• El-Naggar S A, Alm-Eldeen AA, Germoush M O, El-Boray K F, Elgebaly H A. 2015. Ameliorative effect of propolis against cyclophosphamide-induced toxicity in mice.Pharm Biol.53(2):235-41. doi: 10.3109/13880209.2014.914230. Epub 2014 Oct 7.

• ErdoganS, Ates B, DurmazG, YilmazI, Seckin T. 2011. Pressurized liquid extraction of phenolic compounds from Anatolia propolis and their radical scavenging capacities. Food ChemToxicol. 49: 1592–1597.

• Fredriksson L, Wink S, Herpers B, Benedetti G, Hadi M, Bont H D, GroothuisG, Luijten M, Danen E, Graauw M D, Meerman J, Bob van de Water. 2014. Drug-Induced Endoplasmic Reticulum and Oxidative Stress Responses Independently Sensitize Toward TNF-α Mediated Hepatotoxicity. Toxicological sciences. 140(1): 144–159.doi: 10.1093/toxsci/kfu072.

• FuY, ZhengS, LinJ, RyerseJ, Chen A. 2008. Curcumin protects the rat liver from CCl4-caused injury and fibrogenesis by attenuating oxidative stress and suppressing inflammation. MolPharmacol. 73 (2):399-409.

• Gowri Shankar N L, Manavalan R, Venkappayya D. 2008. Hepatoprotective and antioxidant effects of Commiphoraberryi (Arn) Engl bark extract against CCl(4)-induced oxidative damage in rats. Food Chem. Toxicol. 46: 3182–3185.

• Hassan A A M, Ismail M F, Mohamed H M.2015. Effect of methotrexate cmbined with ginger, Silymarin and propolis on mRNA expression levels of cytochrome P45 oxidoreductase (POR), Caspase 3 (CASP-3) and iterlukin 6 9IL-6). Afr. J. Biotechnol. 14(8): 695-701.

• HemeidaR A,MohafezO M. 2008. Curcumin attenuates methotraxate-induced hepatic oxidative damage in rats. J Egypt Natl Canc Inst. 20(2):141–8.

• Jahovic N, CevikH, SehirliAO, YˇegenBC,Sener G. 2003. Melatonin prevents methotraxate- induced hepatorenal oxidative injury in rats. J Pineal Res. 34: 282-287.

• Jasprica D, Mornar A, Debelijak Z, Smolcic-Bubalo A, Medic-Saric M, Mayer L, Romic Z, Bucan K, Balog T, Sobocanec S, SverkoV.2007. In vivo study of propolis supplementation effects on antioxidative status and red blood cells. J. Ethnopharmacol. 110: 548–554.

• Johansen J S, Harris A K, Rychly D, Ergu A. 2005.Oxidative stress and the use of antioxidants in diabetes: Linking basic science to clinical practice. Cardiovascular Diabetology. 4:5 doi:10.1186/1475-2840-4-5.

• Kivity S, Zafrir Y, Loebstein R, Pauzner R, Mouallem M. Mayan H. 2014. Clinical characteristics and risk factors for low dose methotrexate toxicity: A cohort of 28 patients. Autoimmunity Reviews. 13(11): 1109–1113.

• Kose E, Sapmaz H I, Sarihan E, Vardi N, Turkoz Y, Ekinci N. 2012. Beneficial effects of montelukast against methotrexate-induced liver toxicity: a biochemical and histological study. The Scientific World Journal. 2012:987508. doi: 10.1100/2012/987508.

• Lu J, Jones A D, Harkema J R, Roth R A, GaneyP E. 2012. Amiodarone exposure during modest inflammation induces idiosyncrasy-like liver injury in rats: role of tumor necrosis factor-alpha. Toxicol. Sci. 125: 126–133.

• Madrigal-Santillán E, Madrigal-Bujaidar E, Ãlvarez-González I, Sumaya-Martínez M T Gutiérrez-Salinas J., Bautista M, Morales-González A, García-Lunay M, González-Rubio J, Aguilar-Faisal L, Morales-González J A. 2014. Review of natural products with hepatoprotective effects. World J Gastroenterol. 20(40): 14787-14804.

• MaitiR, Jana D, Das U K, Ghosh D. 2004. Antidiabetic effect of aqueous extract of seed of Tamarindusindica in streptozotocin-induced diabetic rats. J Ethnopharmacol. 92: 85-91.

• Moghadam A R, Tutunchi S, Namvaran-Abbas-Abad A, Yazdi M, Bonyadi F, MohajeriD, Mazani M,Marzban H, Åos M J, Ghavami S. 2015. Pre-administration of turmeric prevents methotrexate-induced liver toxicity and oxidative stress BMC Complementary and Alternative Medicine.15:246-258.

• Minami M, Yoshikawa H. 1979. A simplified assay method of super oxide dismutase. Clin. Chim. Acta, 29: 337-342.

• MossaAT, HeikalTM, BelaibaM, Raoelison EG, FerhoutH, Bouajila J. 2015.Antioxidant activity and hepatoprotective potential of Cedrelopsisgrevei on cypermethrin induced oxidative stress and liver damage in male mice.BMC Complement Altern Med.15:251. doi: 10.1186/s12906-015-0740-2.

• Nikolova P, Softova E., Kavaldzhieva B. 1994. The functional and morphological changes in the liver and kidneys of white rats treated with aluminum. Eksp. Med. Morfol. 32: 52–61.

• Oršolić N, Bašić I. 2005. Antitumor, hematostimulative and radioprotective action of water-soluble derivative of propolis (WSDP) Biomed Pharmacother. 2005;59:561–570. doi: 10.1016/j.biopha.2005.03.013.

• Pepys M B, Hirschfield G M. 2003. C-reactive protein: a critical update. J Clin Invest. 111: 1805-12.

• Retiman S, Frankel S.1957. Colorimetric method for the determination of serum transaminase activity. Am. J. Clin. Path. 28: 56-59.

• Rizk S M, Zaki H F, Mina M A. 2014. Propolis attenuates doxorubicin-induced testicular toxicity in rats, Food Chem. Toxicol. 67: 176–186.

• Rosen H R, Keeffe H R. 2000. Evaluation of abnormal liver enzymes, use of liver test and the serology of viral hepatitis, In: Bacon BR, Bisceglie AM. Liver Disease. Diagnosis and management, New York, Churchill Livingstone. 24-35.

• Roth R A, Ganey P E. 2011. Animal models of idiosyncratic druginduced liver injury—current status. Crit. Rev. Toxicol. 41: 723–739.

• Salas A L, AlbertoMR,Zampini I C, CuelloAS, MaldonadoL, Ríos JL, Schmeda-HirschmannG,Isla MI. 2016. Biological activities of polyphenols-enriched propolis from Argentina arid regions.Phytomedicine.23(1):27-31.

• Schmiegelow, K. (2009). Advances in individual prediction of methotrexate toxicity: A review. Br. J. Haematol. 146: 489–503.

• Sener G, Ekşioğlu-Demiralp E, Cetiner M, Ercan F, Yeğen B C. 2006. Beta-glucan ameliorates methotrexate-induced oxidative organ injury via its antioxidant and immunomodulatory effects. Eur J Pharmacol. 542(1-3):170-8.

• Sönmeza M F, Çilenka K T, Karabuluta D, Ünalmışa S, Deligönülb E, Öztürkc I, Kaymaka E. 2016. Protective effects of propolis on methotrexate-induced testis injury in rat. Biomedicine & Pharmacotherapy 79. 44–51.

• Swelm V R P, Laarakkers C M, Kooijmans-Otero M, de Jong E M, Masereeuw R, Russel F G. 2013. Biomarkers for methotrexate-induced liver injury: Urinary protein profiling of psoriasis patients. Toxicol Lett. 221(3):219–24.

• Talas ZS, Dundar SP, Gulhan MF, Orun I,Kakoolaki S. 2012. Effects of propolis on some blood parameters and enzymes in carp exposed to arsenic. Iran J Fish Sci. 11: 405–414.

• Tian H, Cronstein B N. 2007. Understanding the mechanisms of action of methotrexate. ull NYU HospJt Dis. 65(3):168–73.

• Wali A F, Avula B, Ali Z, Khan IA, MushtaqA, RehmanMU, Akbar S, MasoodiMH. 2015. Antioxidant, Hepatoprotective Potential and Chemical Profiling of Propolis Ethanolic Extract from Kashmir Himalaya Region Using UHPLC-DAD-QToF-MS.Biomed Res Int. 2015:393462. doi: 10.1155/2015/393462.

• Whitfield J B, Moss D W, Neale G, Orme M, Breckenridge A. 1973. Changes in plasma -glutamyltranspeptidase activity associated with alterations in drug metabolism in man.Br. Med. J., 1 (5849): 316-318.

• WidedK, HassibaR, MesbahL. 2014.Polyphenolic fraction of Algerian propolis reverses doxorubicin induced oxidative stress in liver cells and mitochondria.Pak J Pharm Sci.27(6):1891-7.




How to Cite

Badr, G. M. (2016). Ameliorative Effect of Propolis Extract on Hepatotoxicity Induced by Methotrexate in Mice. Asian Journal of Applied Sciences, 4(4). Retrieved from