OPEN-ACCESS PEER-REVIEWED
1Dhanya Shrijith Pillai, 2Bhalchandra Baburao Waykar
1Department of Zoology, Dr. Babasaheb Ambedkar Marathwada University, Chhatrapati Sambhajinagar-431004, Maharashtra, India
2Department of Zoology, Dr. Babasaheb Ambedkar Marathwada University, Chhatrapati Sambhajinagar-431004, Maharashtra, India
Abstract
Methotrexate, an antifolate drug is used to cure various inflammatory illness and cancer at different stages. Although it is an effective drug it has the potency to cause toxic effects on majority of the body organs. Natural antioxidants like honey and bee venom have curative properties against the side effects of the drug. The objective of the research is to examine the short-term immediate response of the drug and the role of honey and bee venom against the drug mediated changes upon the hepatic and renal function biomarkers of wistar albino rats. Rats about 10-12 weeks age and weighing around 200-250g were randomly allocated into four groups. Group 1:(Normal Control): Rats were administered 0.9% saline. Group II (Disease control): Rats were delivered intraperitoneally with methotrexate (300 µg/kg.bwt. /day) for 15 days. Group 111(Test group 1): Rats were orally administered with honey (500mg/kg.bwt/day)following bee venom(0.5mg/kg)injected intraperitoneal for 15 days. Group 1V:(Test group II):Rats were administered methotrexate 300µg/kg b.wt/day) and bee venom daily( 0.5mg/kg )injected intraperitoneally with honey orally fed(500mg/kg bwt/day) through intragastric tube for about 15 days. After concluding the experiment, blood was drawn for serum separation through centrifugation at 3500 rpm for the analysis of liver function parameters like ALT, AST, ALP, Total bilirubin, albumin and kidney function indicators like creatinine, urea and uric acid. Both the organs were subjected to histopathological examination. On comparing the control group with the disease group, the administration of methotrexate to the rats (G-11) led to a significant increase in the levels of ALT, AST, ALP, Bilirubin levels whereas there was a considerable fall in the albumin level. There was also significant renal damage indicated by an increase in the levels of creatinine, urea and uric acid in the disease group. In both of the test groups, group III and group 1V the supplementation of honey plus bee venom and the supplementation of methotrexate with honey and bee venom significantly lowered the enzyme activities of ALT,AST,ALP, and bilirubin levels with an increase in the albumin level compared to disease group. Both of the test groups Group III and Group IV significantly lowered the values of creatinine, urea and uric acid than the disease group. The normal control and both of the test groups had highly significant changes when compared to the disease control group. The present study found that the combined administration of honey and bee venom of Apis mellifera against methotrexate mediated changes in liver and kidney function biomarkers has curative role due to its potent antioxidant, free radical scavenging and anti-oxidative activities and hence recommended against the toxic effects of the drug on different physiological aspects of the body.
Keywords: Wistar male albino rats, liver function, kidney function, methotrexate, honey, bee venom
References
[1]. Puig L (2014): Methotrexate: new therapeutic approaches. Actas Dermosifiliogr 105: 583-589. Proinflammatory cytokines (TNF-alpha and IL-1beta) production by water-soluble sub-fractionated parts from bee (Apis Pharmazie, 56: 239-241.
[2]. Tousson, Ehab, et al. “Abrogation by Ginkgo Byloba leaf extract on hepatic and renal toxicity induced by methotrexate in rats.” Journal of Cancer Research and Treatment 2.3 (2014): 44-51.
[3]. Jolivet J, Cowan KH, Curt GA, Clendeninn NJ, Chabner BA. The pharmacology and clinical use of methotrexate. N Engl J Med 1983;309:1094‑104..
[4]. Cronstein, Bruce N., and Thomas M. Aune. “Methotrexate and its mechanisms of action in inflammatory arthritis.” Nature Reviews Rheumatology 16.3 (2020): 145-154.
[5]. Kamen, B. A., et al. “Methotrexate accumulation and folate depletion in cells as a possible mechanism of chronic toxicity to the drug.” British journal of haematology 49.3 (1981): 355-360.
[6]. Gibson, Rachel J., and Joanne M. Bowen. “Biomarkers of regimen-related mucosal injury.” Cancer treatment reviews 37.6 (2011): 487-493.
[7]. Argyriou AA, et al. (2012): Chemotherapy-induced peripheral neurotoxicity (CIPN): an update. Crit Rev Oncol Hematol. 2012; 8
[8]. Blumenfeld Z. (2012): Chemotherapy and fertility. Best Pract Res Clin Obstet Gynaecol. 2012; 26(3):379–90
[9]. Mohamed, Hanaa K., et al. “Anti-inflammatory, anti-apoptotic, and antioxidant roles of honey, royal jelly, and propolis in suppressing nephrotoxicity induced by doxorubicin in male albino rats.” Antioxidants 11.5 (2022): 1029.
[10]. Kintzel, Polly E. “Anticancer Drug—Induced Kidney Disorders: Incidence, Prevention and Management.” Drug safety 24.1 (2001): 19-38.
[11]. Younis, Nancy S., et al. “Geraniol averts methotrexate-induced acute kidney injury via Keap1/Nrf2/HO-1 and MAPK/NF-κB pathways.” Current Issues in Molecular Biology 43.3 (2021): 1741-1755.
[12]. Mahmoud, Ayman M., et al. “Commiphora molmol protects against methotrexate-induced nephrotoxicity by up-regulating Nrf2/ARE/HO-1 signaling.” Biomedicine & Pharmacotherapy 106 (2018): 499-509.
[13]. Jafaripour, Leila, et al. “Effects of rosmarinic acid on methotrexate-induced nephrotoxicity and hepatotoxicity in wistar rats.” Indian journal of nephrology 31.3 (2021): 218-224.
[14]. Hassan, Osama Abdelaziz, Entesar Farghally Amin, and Rabab Ahmed Moussa. “Protective effect of erdosteine against methotrexateinduced hepatotoxicity in rats.” Tropical Journal of Pharmaceutical Research 19.7 (2020): 1465-1471.
[15]. Al-Motabagani, Mohamed Akram. “Histological and histochemical studies on the effects of methotrexate on the liver of adult male albino rat.” Int J Morphol 24.3 (2006): 417-22.
[16]. Quintin, Emilie, et al. “Rare incidence of methotrexate-specific lesions in liver biopsy of patients with arthritis and elevated liver enzymes.” Arthritis research & therapy 12 (2010): 1-7.
[17]. El-Sheikh, Azza AK, et al. “Mechanisms of thymoquinone hepatorenal protection in methotrexate-induced toxicity in rats.” Mediators of inflammation 2015 (2015).
[18]. Sahindokuyucu-Kocasari, F., et al. “Apigenin alleviates methotrexate-induced liver and kidney injury in mice.” Human & Experimental Toxicology 40.10 (2021): 1721-1731
[19]. Gerber, M.; Boutron-Ruault, M. C.; Hercberg, S.; Riboli, E.; Scalbert, A. and et al. (2002): Food and cancer: state of the art about the protective effect of fruits and vegetables. Bull Cancer, 89: 293-312.
[20]. Helen, N. Saada, and S. Azab Khaled. “Role of lycopene in recovery of radiation induced injury to mammalian cellular organelles.” Die Pharmazie 56.3 (2001): 239-241.
[21]. Son, Dong Ju, et al. “Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds.” Pharmacology & therapeutics 115.2 (2007): 246-270.
[22]. Kang, Seong Soo, Sok Cheon Pak, and Seok Hwa Choi. “The effect of whole bee venom on arthritis.” The American journal of Chinese medicine 30.01 (2002): 73-80.
[23]. Kim, Hyunseong, et al. “Bee venom melittin protects against cisplatin-induced acute kidney injury in mice via the regulation of M2 macrophage activation.” Toxins 12.9 (2020): 574.
[24]. Somwongin, Suvimol, Panuwan Chantawannakul, and Wantida Chaiyana. “Antioxidant activity and irritation property of venoms from Apis species.” Toxicon 145 (2018): 32-39.
[25]. Varanda, Eliana Ap, Rubens Monti, and Denise C. Tavares. “Inhibitory effect of propolis and bee venom on the mutagenicity of some direct‐and indirect‐acting mutagens.” Teratogenesis, carcinogenesis, and mutagenesis 19.6 (1999): 403-413.
[26]. Sumikura, Hiroyuki, et al. “A comparison of hyperalgesia and neurogenic inflammation induced by melittin and capsaicin in humans.” Neuroscience letters 337.3 (2003): 147-150.
[27]. Nam, Kung-Woo, et al. “Inhibition of COX-2 activity and proinflammatory cytokines (TNF-α and IL-1β) production by water-soluble sub-fractionated parts from bee (Apis mellifera) venom.” Archives of Pharmacal Research 26 (2003): 383-388.
[28]. Kim, Hyun-Woo, et al. “Acupoint stimulation using bee venom attenuates formalin-induced pain behavior and spinal cord fos expression in rats.” Journal of veterinary medical science 65.3 (2003): 349-355.
[29]. Orsolic, Nada, and Ivan Basic. “APOPTOSIS AND NECROSIS AS POSSIBLE MECHANISMS FOR ANTITUMOR ACTIVITY OF BEE VENOM.” Mellifera 3.5 (2003).
[30]. Gajski, G. and Garaj-Vrhovac, V. (2009): Radioprotective effects of honey bee venom (Apis mellifera) against 915-MHz microwave radiation-induced DNA damage in Wistar rat lymphocytes: in vitro study. Int. J. Toxicol., 28(2): 88-98.
[31]. Choi, Gwang-Muk, et al. “Bee venom phospholipase A2 alleviates collagen-induced polyarthritis by inducing Foxp3+ regulatory T cell polarization in mice.” Scientific Reports 11.1 (2021): 3511.
[32]. Goo, Bonhyuk, et al. “Bee venom alleviated edema and pain in monosodium urate crystals-induced gouty arthritis in rat by inhibiting inflammation.” Toxins 13.9 (2021): 661.
[33]. Conrad, Vicki J., et al. “Efficacy and safety of honey bee venom (Apis mellifera) dermal injections to treat osteoarthritis knee pain and physical disability: A randomized controlled trial.” The Journal of Alternative and Complementary Medicine 25.8 (2019): 845-855.
[34]. Khalil, M. I., Siti Amrah Sulaiman, and Laïd Boukraa. “Antioxidant properties of honey and its role in preventing health disorder.” The open nutraceuticals journal 3.1 (2010).
[35]. Bhalchandra, Waykar, and Yahya Ali Alqadhi. “Administration of honey and royal jelly ameliorate cisplatin induced changes in liver and kidney function in rat.” Biomedical and Pharmacology Journal 11.4 (2018): 2191-2199.
[36]. Erejuwa, O. O., et al. “Effects of Malaysian tualang honey supplementation on glycemia, free radical scavenging enzymes and markers of oxidative stress in kidneys of normal and streptozotocin-induced diabetic rats.” International Journal of Cardiology 137 (2009): S45.
[37]. Neamatallah, Thikryat, et al. “Honey protects against cisplatin-induced hepatic and renal toxicity through inhibition of NF-κB-mediated COX-2 expression and the oxidative stress dependent BAX/Bcl-2/caspase-3 apoptotic pathway.” Food & function 9.7 (2018): 3743-3754.
[38]. Hamad, Rania, et al. “Honey feeding protects kidney against cisplatin nephrotoxicity through suppression of inflammation.” Clinical and Experimental Pharmacology and Physiology 42.8 (2015): 843-848.
[39]. Hall, Pauline de la M., Mark A. Jenner, and Michael J. Ahern. “Hepatotoxicity in a rat model caused by orally administered methotrexate.” Hepatology 14.5 (1991): 906-910.
[40]. Mousavi, Seyyedeh Mahbubeh, et al. “Effect of Iranian honey bee (Apis mellifera) venom on blood glucose and insulin in diabetic rats.” Journal of arthropod-borne diseases 6.2 (2012): 136.
[41]. Ibrahim, Abdelazim, Mabrouk A. Abd Eldaim, and Mohamed M. Abdel-Daim. “Nephroprotective effect of bee honey and royal jelly against subchronic cisplatin toxicity in rats.” Cytotechnology 68.4 (2016): 1039-1048.
[42]. Abd Ali,Abeer R.,and Sajida H.Ismail.”The protective effect of honey against amikacin-induced nephrotoxicity in rats.Iraqi J Pharm Sci 21.2(20120:85-93
[43]. Roghani, Mozhdeh, et al. “Alleviation of liver dysfunction, oxidative stress and inflammation underlies the protective effect of ferulic acid in methotrexate-induced hepatotoxicity.” Drug design, development and therapy (2020): 1933-1941.
[44]. Karlsson Sundbaum, Johanna, et al. “Methotrexate treatment in rheumatoid arthritis and elevated liver enzymes: A long‐term follow‐up of predictors, surveillance, and outcome in clinical practice.” International journal of rheumatic diseases 22.7 (2019): 1226-1232.
[45]. Kremer, Joel M., et al. “Methotrexate metabolism analysis in blood and liver of rheumatoid arthritis patients: association with hepatic folate deficiency and formation of polyglutamates.” Arthritis & Rheumatism: Official Journal of the American College of Rheumatology 29.7 (1986): 832-835.
[46]. Bedoui, Yosra, et al. “Methotrexate an old drug with new tricks.” International journal of molecular sciences 20.20 (2019): 5023
[47]. Ezhilarasan, Devaraj. “Hepatotoxic potentials of methotrexate: Understanding the possible toxicological molecular mechanisms.” Toxicology 458 (2021): 152840.
[48]. Vaghasiya, Jitendra, Yagnik Bhalodia, and Shivkumar Rathod. “Drug induced hepatotoxicity: effect of polyherbal formulation.” Pharmacognosy Magazine 5.19 (2009).
[49]. Giannini, Edoardo G., and Markus Peck-Radosavljevic. “Platelet dysfunction: status of thrombopoietin in thrombocytopenia associated with chronic liver failure.” Seminars in Thrombosis and Hemostasis. Thieme Medical Publishers, 2015.
[50]. Walker, T. M., P. C. Rhodes, and C. Westmoreland. “The differential cytotoxicity of methotrexate in rat hepatocyte monolayer and spheroid cultures.” Toxicology in vitro 14.5 (2000): 475-485.
[51]. Chauhan, Prerna, et al. “Protective effects of Glycyrrhiza glabra supplementation against methotrexate-induced hepato-renal damage in rats: An experimental approach.” Journal of Ethnopharmacology 263 (2020): 113209.
[52]. Dalaklioglu, S. E. L. V. İ. N. A. Z., et al. “Resveratrol ameliorates methotrexate-induced hepatotoxicity
[53]. Vardi, Nigar, et al. “Protective effect of β-carotene on methotrexate–induced oxidative liver damage.” Toxicologic pathology 38.4 (2010): 592-597.
[54]. Hersh, Evan M., et al. “Hepatotoxic effects of methotrexate.” Cancer 19.4 (1966): 600-606.
[55]. Sakeran, Mohamed I., et al. “Abrogation by Trifolium alexandrinum root extract on hepatotoxicity induced by acetaminophen in rats.” Redox Report 19.1 (2014): 26-33.
[56]. ŞENER, GÖKSEL. “Amelioration of methotrexate-induced enteritis by melatonin in rats.” (2004).
[57]. Drotman, R. B., and G. T. Lawhorn. “Serum enzymes as indicators of chemically induced liver damage.” Drug and chemical toxicology 1.2 (1978): 163-171.
[58]. Kadikoylu, G., et al. “The effects of desferrioxamine on cisplatininduced lipid peroxidation and the activities of antioxidant enzymes in rat kidneys.” Human & experimental toxicology 23.1 (2004): 29-34.
[59]. Reiss, Samantha N., et al. “Hypoalbuminemia is significantly associated with increased clearance time of high dose methotrexate in patients being treated for lymphoma or leukemia.” Annals of hematology 95 (2016): 2009-2015.
[60]. Mohassel, Leila, et al. “Evaluation of Methotrexate Clearance in Adult and Pediatric Patients with Hypoalbuminemia.” Blood 134 (2019): 2906.
[61]. Slouma, Maroua, et al. “Associated factors with liver fibrosis in rheumatoid arthritis patients treated with methotrexate.” Clinical Rheumatology 43.3 (2024): 929-938.
[62]. Furuya, Takefumi, et al. “Prevalence of high and low serum alkaline phosphatase levels and the associated factors in patients with rheumatoid arthritis: Results from the IORRA cohort study.” Modern Rheumatology (2024): roae025.
[63]. Mahmoud, Ayman M., et al. “Methotrexate hepatotoxicity is associated with oxidative stress, and down-regulation of PPARγ and Nrf2: Protective effect of 18β-Glycyrrhetinic acid.” Chemico-biological interactions 270 (2017): 59-72.
[64]. Hadi, Najah R., Fadhil G. Al-Amran, and Asma Swadi. “Metformin ameliorates methotrexate-induced hepatotoxicity.” Journal of Pharmacology and Pharmacotherapeutics 3.3 (2012): 248-253.
[65]. Meligi, Noha M., Suzan Alaa Ismail, and Nagy S. Tawfik. “Protective effects of honey and bee venom against lipopolysaccharide and carbon tetrachloride-induced hepatoxicity and lipid peroxidation in rats.” Toxicology Research 9.5 (2020): 693-705.
[66]. Naji, Khalid Mohammed, et al. “Hepatoprotective activity of melittin on isoniazid-and rifampicin-induced liver injuries in male albino rats.” BMC Pharmacology and Toxicology 22.1 (2021): 39.
[67]. Lee, Woo-Ram, Sok Cheon Pak, and Kwan-Kyu Park. “The protective effect of bee venom on fibrosis causing inflammatory diseases.” Toxins 7.11 (2015): 4758-4772.
[68]. Kim, Jung-Yeon, Jaechan Leem, and Hyo-Lim Hong. “Melittin ameliorates endotoxin-induced acute kidney injury by inhibiting inflammation, oxidative stress, and cell death in mice.” Oxidative medicine and cellular longevity 2021 (2021): 1-14
[69]. Onochie, Maureen, Chuemere Arthur Nwafor, and Ilochi Ogadinma. “Hepatoprotective potential of honey, coffee and vitamin E in male wistar rats.” Eur J Pharm Sci 5 (2018): 47-51.
[70]. Muhammad, M. M. A., M. Mouchira, and R. A. Naglaa. “Physiological effects of bee venom and propolis on irradiated albino rats.” Danish J Agricult Animal Sci 2015 (2015): 11-21.
[71]. Glantzounis, G. K., et al. “Uric acid and oxidative stress.” Current pharmaceutical design 11.32 (2005): 4145-4151.
[72]. Asci, Halil, et al. “The impact of gallic acid on the methotrexate-induced kidney damage in rats.” journal of food and drug analysis 25.4 (2017): 890-897Mahmoud, Ayman M., et al. “Commiphora molmol protects against methotrexate-induced nephrotoxicity by up-regulating Nrf2/ARE/HO-1 signaling.” Biomedicine & Pharmacotherapy 106 (2018): 499-509.
[73]. Kanbay, Mehmet, et al. “The role of uric acid in the pathogenesis of human cardiovascular disease.” Heart 99.11 (2013): 759-766.
[74]. Kobayashi, Takehito, et al. “Elevated uric acid and adenosine triphosphate concentrations in bronchoalveolar lavage fluid of eosinophilic pneumonia.” Allergology International 66.Supplement. 1 (2017): S27-S34.
[75]. Kawaguchi, Shinichiro, et al. “Risk factors for high-dose methotrexate-induced nephrotoxicity.” International Journal of Hematology 114 (2021): 79-84.
[76]. Yang, Shi-Long, et al. “Methotrexate associated renal impairment is related to delayed elimination of high-dose methotrexate.” The Scientific World Journal 2015 (2015).
[77]. Howard, Scott C., et al. “Preventing and managing toxicities of high-dose methotrexate.” The oncologist 21.12 (2016): 1471-1482.
[78]. Heidari, Reza, et al. “Mitochondrial dysfunction and oxidative stress are involved in the mechanism of methotrexate-induced renal injury and electrolytes imbalance.” Biomedicine & Pharmacotherapy 107 (2018): 834-840.
[79]. Elmansy, Rasha A., et al. “Rebamipide potentially mitigates methotrexate‐induced nephrotoxicity via inhibition of oxidative stress and inflammation: A molecular and histochemical study.” The Anatomical Record 304.3 (2021): 647-661.
[80]. Devrim, Erdinç, et al. “Methotrexate causes oxidative stress in rat kidney tissues.” Renal failure 27.6 (2005): 771-773.
[81]. Donate-Correa, Javier, et al. “Klotho, oxidative stress, and mitochondrial damage in kidney disease.” Antioxidants 12.2 (2023): 239.
[82]. Zaaba, Nur Elena, et al. “Catalpol attenuates oxidative stress and inflammation via mechanisms involving sirtuin-1 activation and NF-κB inhibition in experimentally-induced chronic kidney disease.” Nutrients 15.1 (2023): 237.
[83]. Gyurászová, Marianna, et al. “Oxidative stress in the pathophysiology of kidney disease: implications for noninvasive monitoring and identification of biomarkers.” Oxidative medicine and cellular longevity 2020 (2020).
[84]. Kunat-Budzyńska, Magdalena, et al. “Chemical composition and antimicrobial activity of new honey varietals.” International Journal of Environmental Research and Public Health 20.3 (2023): 2458.
[85]. Jaganathan, Saravana Kumar, and Mahitosh Mandal. “Antiproliferative effects of honey and of its polyphenols: a review.” BioMed Research International 2009 (2009).
[86]. Kim, Jung-Yeon, Jaechan Leem, and Kwan-Kyu Park. “Antioxidative, antiapoptotic, and anti-inflammatory effects of apamin in a murine model of lipopolysaccharide-induced acute kidney injury.” Molecules 25.23 (2020): 5717.
[87]. Kim, Jung-Yeon, et al. “Protective effects of bee venom against endotoxemia-related acute kidney injury in mice.” Biology 9.7 (2020): 154.
[88]. Elbakry, KADRY A., CAMELIA A. ABDEL Malak, and MAHMOUD M. Howas. “Immunomodulatory role of honey and propolis on carbon tetrachloride (CCl4) injected rats.” Int J Pharm Pharm Sci 7.12 (2015): 259-262.
[89]. Castro, Henry J., et al. “A phase I study of the safety of honeybee venom extract as a possible treatment for patients with progressive forms of multiple sclerosis.” Allergy & Asthma Proceedings. Vol. 26. No. 6. 2005.
[90]. El Adham, Eithar K., Amal I. Hassan, and M. M. A. Dawoud. “Evaluating the role of propolis and bee venom on the oxidative stress induced by gamma rays in rats.” Scientific Reports 12.1 (2022): 2656.
[91]. Osama, Hasnaa, et al. “Effect of honey and royal jelly against cisplatin-induced nephrotoxicity in patients with cancer.” Journal of the American college of nutrition 36.5 (2017): 342-346.
[92]. Muhammad, M. M. A., M. Mouchira, and R. A. Naglaa. “Physiological effects of bee venom and propolis on irradiated albino rats.” Danish J Agricult Animal Sci 2015 (2015): 11-21.
[93]. Eteraf-Oskouei T, Najafi M. Traditional and modern uses of natural honey in human diseases: a review. Iranian journal of basic medical sciences. 2013 Jun;16(6):731.
[94]. Abd El-Rahim, Abeer H., et al. “Inhibitory effect of bee venom against potassium bromate causing genetic toxicity and biochemical alterations in mice.” Journal of The Arab Society for Medical Research 13.2 (2018): 89-98.
[95]. Aouadi L, Dahdouh F, Djebar–berrabeh H. Possible curative effect of venom collected from Algerian bees (Apis mellifera intermissa) on adenine-induced chronic kidney damage in mice. Egyptian Journal of Basic and Applied Sciences. 2024 Dec 31;11(1):135-47.
[96]. Darwish, Samar F., et al. “Targeting TNF-α and NF-κB activation by bee venom: role in suppressing adjuvant induced arthritis and methotrexate hepatotoxicity in rats.” PLoS One 8.11 (2013): e79284.
[97]. El-Sheikh, Azza AK, et al. “Mechanisms of thymoquinone hepatorenal protection in methotrexate-induced toxicity in rats.” Mediators of inflammation 2015 (2015).
[98]. Sahindokuyucu-Kocasari, F., et al. “Apigenin alleviates methotrexate-induced liver and kidney injury in mice.” Human & Experimental Toxicology 40.10 (2021): 1721-1731.
[99]. Al-Abkal, Faten, et al. “Protective effect of pycnogenol against methotrexate-induced hepatic, renal, and cardiac toxicity: An in vivo study.” Pharmaceuticals 15.6 (2022): 674.
[100]. Fadel, Maab A., et al. “Protective effect of propolis on liver and kidney injury caused by methotrexate in chicks.” (2022): 1061-1067.
[101]. Erdogan, Esra, et al. “Rutin ameliorates methotrexate induced hepatic injury in rats.” Acta cirurgica brasileira 30 (2015): 778-784.
[102]. Abdel-Moneim, Wafaa M., and Hemmat H. Ghafeer. “The potential protective effect of natural honey against cadmium-induced hepatotoxicity and nephrotoxicity.” Mansoura Journal of Forensic Medicine and Clinical Toxicology 15.2 (2007): 75-98.
[103]. Korkmaz, Aslı, and Dürdane Kolankaya. “Anzer honey prevents N-ethylmaleimide-induced liver damage in rats.” Experimental and Toxicologic Pathology 61.4 (2009): 333-337.
[104]. Halawa, Heba M., et al. “Evaluation of honey protective effect on lead induced oxidative stress in rats.” Jasmr 4.2 (2009): 197-209.
[105]. Al-AwarM S A, Gumaih H S A and Al-Ameri D A A. The protective effect of Sider honey and Zinc on imidacloprid induced hepatorenal and hematological toxicity in rats. Journal of Natural Sciences, Life and Applied sciences AJSRP. 2018: (2): (1)
[106]. El-Badawi, M. G., et al. “Histological changes following high-dose methotrexate and cisplatinum administration and the influence of dosage scheduling.” Chemotherapy 33.4 (1987): 278-286.
[107]. Wadi, Siham A., et al. “PROTECTIVE EFFECT OF PUMPKIN SEED OIL ON METHOTREXATE-INDUCED NEPHROTOXICITY IN RATS.” Biochemical & Cellular Archives 21.2 (2021).
[108]. Rizk, Fatma H., et al. “Metformin ameliorated methotrexate-induced hepatorenal toxicity in rats in addition to its antitumor activity: two birds with one stone.” Journal of Inflammation Research (2018): 421-429.
[109]. Alharbi, Fahad Hamadan M., et al. “Severe Renal Impairment in a Patient with Recent Rheumatoid Arthritis Diagnosis following Methotrexate Initiation: A Case Report.” Journal of Pharmacy and Bioallied Sciences 16.Suppl 2 (2024): S1878-S1882.
[110]. Wasfey, Eman F., et al. “Infliximab Ameliorates Methotrexate-Induced Nephrotoxicity in Experimental Rat Model: Impact on Oxidative Stress, Mitochondrial Biogenesis, Apoptotic and Autophagic Machineries.” Cell Biochemistry and Biophysics 81.4 (2023): 717-726.
[111]. Shi, Peiying, et al. “Pharmacological effects and mechanisms of bee venom and its main components: recent progress and perspective.” Frontiers in pharmacology 13 (2022): 1001553.