Experimental animal Biology

In collaboration with Payame Noor University and Iranian Society of Physiology and Pharmacology

Current Issue

By Issue

By Author

By Subject

Author Index

Keyword Index

About Journal

Aims and Scope

Editorial Board

Publication Ethics

Indexing and Abstracting

Related Links

FAQ

Peer Review Process

Journal Metrics

News

The Influences of Hydro-Alcoholic Extract of Melilotus officinalis ‎on Blood Liver Enzymes in Adult Male Rats Under the Chronic ‎Stress

Document Type : Article

Authors

1 Department of Biology, Faculty of ‎Basic Sciences, Shahrekord ‎University, Shahrekord, Iran‎

2 Department of Biology, Iz.C., Islamic ‎Azad University, Izeh, Iran

10.30473/eab.2026.77209.2028

Abstract

The liver enzymes are the liver health indicators in which their increase uncovers the liver damage. This experimental research was designed to examine the hydro-alcoholic extract' effect of M. officinalis plant on blood liver enzymes in the adult male rats, under the chronic stress. To do so, 40 male Wistar rats were randomly classified into the control, vehicle-treated, and three experimental groups that received chronic stress and treated with dosages of 25, 50 and 100 (mg/Kg) of M. officinalis extract. To induce chronic immobility stress, animals were daily placed in the restraint device for two hrs during 21 days. The plant extract was injected intraperitoneally 30 minutes before stress induction. After the deep anesthesia and blood sampling from the animals' hearts, the blood serum samples were taken to evaluate the liver enzymes' level. In the vehicle-treated group, AST, ALT, and ALP levels showed significant increases compared to the control group (p<0.05, p<0.01, p<0.001, respectively). The M. officinalis extract at 50 mg/kg significantly reduced AST and ALP levels compared to the vehicle-treated group (p<0.001, p<0.05) and significantly reduced ALT levels compared to both vehicle-treated and control groups (p<0.001, p<0.05). Administration of 25 and 100 mg/kg of the extract significantly lowered ALT levels relative to the vehicle-treated group (p<0.001, p<0.01), while no significant changes were observed in the other two enzymes.As a conclusion, the medium dose of M. officinalis extract could reduce high level of the liver blood enzymes in this stress.

Keywords

Main Subjects

References
Ahmadi, A., Servat-Yari, K., Kashefi, H., Monbari, M. N., Rostami, A., & Moloudi, M. R. (2016). The effect of hydroalcoholic extract of alfalfa on liver function tests, blood biochemical factors and coagulation system in male rats. Scientific Journal of Kurdistan University of Medical Sciences, 21(6), 16-26.
Ahsan, H., Khan, K. U., Khan, I. U., Saleem, M., Khadija, H. A., & Sharif, A. (2017). Evaluation of hepatoprotective activity of Melilotus officinalis L. against paracetamol and carbon tetrachloride induced hepatic injury in mice. Acta Poloniae Pharmaceutica – Drug Research, 74(3), 903–909.
Avci, G., Kupeli, E., Eryavuz, A., Yesilada, E., & Kucukkurt, I. (2006). Antihypercholesterolaemic and antioxidant activity assessment of some plants used as remedy in Turkish folk medicine. Journal of Ethnopharmacology,107(3),418–423.
Bajaj, J. S., Ellwood, M., Ainger, T., Burroughs, T., Fagan, A., Gavis, E. A., …, & Wade, J. B. (2017). Mindfulness-based stress reduction therapy improves patient and caregiver-reported outcomes in cirrhosis.Clinical and Translational Gastroenterology, 8(7), e108.
Bathaie, S. Z., & Mousavi, S. Z. (2010). New applications and mechanisms of action of saffron and its important ingredients. Critical Reviews in Food Science and Nutrition,50(8),761–786.
Bradic, J., Petrovic, A., Nikolic, M., Nedeljkovic, N., Andjic, M., Kladar, N., ..., & Kocovic, A. (2024). Newly Developed Semi-Solid Formulations Containing Mellilotus officinalis Extract: Characterization, Assessment of Stability, Safety, and Anti-Inflammatory Activity. Pharmaceutics, 16(8), 1003.
Bradic, J., Petrovic, A., Nikolic, M., Nedeljkovic, N., Andjic, M., Kladar, N., Bolevich, S., Jakovljevic, V., & Kocovic, A. (2024). Newly developed semi-solid formulations containing Melilotus officinalis extract: Characterization, assessment of stability, safety, and anti-inflammatory activity. Pharmaceutics, 16(8), 1003.
Bunney, S., Stodola, J., Volák, J., & Severa, F. (1992). The illustrated encyclopedia of herbs: Their medicinal and culinary uses. Chancellor Press.
Chorepsima, S., Tentolouris, K., Dimitroulis, D., & Tentolouris, N. (2013). Melilotus: Contribution to wound healing in the diabetic foot. Journal of Herbal Medicine, 3(3), 81–86.
Dröge, W. (2002). Free radicals in the physiological control of cell function. Physiological Reviews, 82(1),47–95.
Furukawa, S., Fujita, T., Shimabukuro, M., Iwaki, M., Yamada, Y., Nakajima, Y., …, & Shimomura, I. (2017). Increased oxidative stress in obesity and its impact on metabolic syndrome. The Journal of Clinical Investigation, 114(12), 1752–1761.
Hassani, M., Soleimani, M., Esmaeilzadeh, E., Zare-Abdollahi, D., & Khorshid, H. R. (2020). Healing influence of Melilotus officinalis herbal extract on experimental autoimmune encephalomyelitis in C57BL/6 mice. Iranian Journal of Pharmaceutical Research, 19(4), 321-332.
Ishtiaq, S. M., Khan, J. A., & Arshad, M. I. (2018). Psychosocial-stress, liver regeneration and weight gain: A conspicuous pathophysiological triad. Cellular Physiology andBiochemistry, 46(1),1–8.
Jasicka-Misiak, I., Makowicz, E., & Stanek, N. (2017). Polish yellow sweet clover (Melilotus officinalis L.) honey, chromatographic fingerprints, and chemical markers. Molecules, 22(1),138.
Joung, J. Y., Cho, J. H., Kim, Y. H., Choi, S. H., & Son, C. G. (2019). A literature review for the mechanisms of stress‐induced liver injury. Brain and Behavior, 9(3), e01235. https://doi.org/10.1002/brb3.1235
Kang, D., Zhao, D., Ryu, S., Guallar, E., Cho, J., Lazo, M., …, & Sung, E. (2020). Perceived stress and non-alcoholic fatty liver disease in apparently healthy men and women. Scientific Reports, 10(1), 38.
Karimi Dehkordi, M., Rezaei, F., & Almasi, M. (2021). Investigating the effect of lemon balm and nail extracts on liver function and physiological changes in rats. Journal of Animal Biology, 14(3), 99-109(in Persian).
Mesquita, M., Pedroso, T. F., Oliveira, C. S., Oliveira, V. A., dos Santos, R. F., Bizzi, C. A., & Pereira, M. E. (2016). Effects of zinc against mercury toxicity in female rats 12 and 48 hours after HgCl2 exposure. EXCLI Journal, 15, 256–271. https://doi.org/10.17179/excli2016-107
Moradi, M., Shakerian, S., & Nikbakht, M. (2019). The effect of eight weeks of high-intensity interval training and crocin consumption on oxidative stress in liver tissue of male rats induced by chronic doxorubicin. Feiz Journal of Kashan University of Medical Sciences, 23(5), 485-494. (in Persian)
Nameni, F., Akbari, G., Hadavand, V., Entezari, H., Molaii, S., & Ghadami, N. (2024). Effect of Melilotus Extract and Exercise on Oxidative Stress and Neuroinflammation in an Alzheimer‘s Mouse Model. Jentashapir Journal of Cellular and Molecular Biology, 15(2), e146344.
Noushad, S., Ahmed, S., Ansari, B., Mustafa, U. H., Saleem, Y., & Hazrat, H. (2021). Physiological biomarkers of chronic stress: A systematic review. International Journal of Health Sciences, 15(5), 46–59.
Pu, X., Lu, C., Yang, X., He, H., Chen, X., Wang, R., ..., & Li, Y. (2025). Unveiling the hepatoprotective mechanisms of Desmodium heterocarpon (L.) DC: Novel flavonoid identification and Keap1/Nrf2 pathway activation. Phytomedicine, 136, 156323.
Raeeszadeh, M., Moradi, M., Ayar, P., & Akbari, A. (2021). The antioxidant effect of Medicago sativa L. (alfalfa) ethanolic extract against mercury chloride (HgCl2) toxicity in rat liver and kidney: an in vitro and in vivo study. Evidence-Based Complementary and Alternative Medicine, 2021, Article ID.
Raeeszadeh, M., Mortazavi, P., & Atashin-Sadafi, R. (2021). The antioxidant, anti-inflammatory, pathological, and behavioural effects of Medicago sativa L. (Alfalfa) extract on brain injury caused by nicotine in male rats. Evidence-Based Complementary and Alternative Medicine, 2021, Article ID.
Roshanas, F., Ashrafi, N., Amin Lari, M., & Talibanzadeh, S. (2018). Investigating the effect of saffron aqueous extract on oxidative stress parameters and important biochemical enzymes of liver tissue in streptozotocin-treated diabetic rats. Journal of Arak University of Medical Sciences, 21(5), 77-87.
Sadeghi, T., Shariati, M., & Mokhtari, M. (2016). The effect of hydroalcoholic extract of Passiflora leaf on liver function tests in adult male rats. Journal of Animal Biology, 8(4), 85-96. (in Persian)
Schepetova, E., Abdurakhmanova, N., Lomteva, N., Kondratenko, E., & Kasimova, S. (2021). Pharmacological effects at the chronic administration of Melilotus officinalis L. extract. IOP Conference Series: Earth and Environmental Science, 848(1), 012345.
Servatyari, K., Ahmadi, A., Kashefi, H., Manbari, M. N., Rostami, A., & Moulodi, M. R. (2017). The effect of hydroalcoholic extract of Medicago sativa on liver function tests, blood biochemical factors and coagulation system in male rats. Scientific Journal of Kurdistan University of Medical Sciences, 21(6), 16–26. (In Persian)
Shen, Y., et al. (2025). Schisandrin C Improves Chronic Stress-Induced Dyslipidemia in Mice by Regulating Pyroptosis and Autophagy Levels. Journal of Microbiology and Biotechnology, 35, e2501041.
Soto-Angona, Ó., Anmella, G., Valdés-Florido, M. J., De Uribe-Viloria, N., Carvalho, A. F., Penninx, B. W., & Berk, M. (2020). Non-alcoholic fatty liver disease (NAFLD) as a neglected metabolic companion of psychiatric disorders: Common pathways and future approaches. BMC Medicine, 18(1), 261.
Unsal, V. (2018). Natural phytotherapeutic antioxidants in the treatment of mercury intoxication—A review. Advanced Pharmaceutical Bulletin, 8(3), 365–376.
Yener, M. D., Çolak, T., Özsoy, Ö. D., & Eraldemir, F. C. (2024). Alterations in Catalase, Superoxide Dismutase, Glutathione Peroxidase and Malondialdehyde Levels in Serum and Liver Tissue Under Stress Conditions. Journal of Istanbul Faculty of Medicine, 87(2), 145-152.
Experimental animal Biology
Volume 14, Issue 3 - Serial Number 55
March 2026
Pages 37-47
Files
Share
How to cite
Statistics