Evaluation of Antidepressant Activity of Ethanolic Extract of Celery Leaves (Apium graveolens L.) Using Behavioral Tests in Male Mice

Nicolaas Panji Wailan Saerang; Min Rahminiwati; Sara Nurmala

Authors

Nicolaas Panji Wailan Saerang Universitas Pakuan
Min Rahminiwati Institut Pertanian Bogor
Sara Nurmala Universitas Pakuan

Keywords:

Apium graveolens, antidepressant, depression, FST, monoamine oxidase-A

Abstract

Depression is a prevalent mental disorder associated with dysregulation of monoaminergic neurotransmission and chronic stress exposure. Apium graveolens L. (celery) leaves have been reported to possess neuroactive properties, including monoamine oxidase A (MAO-A) inhibition, suggesting potential antidepressant activity. The objective of this study was to evaluate the antidepressant-like effects of a 70% ethanolic extract of Apium graveolens leaves (EDS) in male Deutch Democratic Yokohama (ddY) mice using the Forced Swim Test (FST) following chronic mild stress (CMS) induction, and to determine an effective dose. Male ddY mice were randomly divided into five groups: negative control, positive control (amitriptyline 5.05 mg/kg body weight), and three EDS-treated groups receiving doses of 100, 150, and 200 mg/kg body weight. The treatments were administered orally in a once-daily. The presence of depressive-like behavior was evaluated through the utilization of the FST, with the duration of immobility being identified as the primary outcome measure. The data were analyzed using one-way analysis of variance (ANOVA) followed by appropriate post hoc tests. The results showed no significant differences in immobility time among groups during the initial treatment phase, indicating that short-term administration did not produce immediate antidepressant-like effects. However, after repeated administration, EDS at a dose of 200 mg/kg body weight significantly reduced immobility time and exhibited an antidepressant-like effect comparable to that of the positive control. Lower doses produced only partial behavioral responses. These findings indicate that the antidepressant-like activity of EDS is both dose-dependent and time-dependent.

References

Alizadeh, A., Pourfallah-Taft, Y., Khoshnazar, M., Safdari, A., Komari, S. V., Zanganeh, M., Sami, N., Valizadeh, M., Faridzadeh, A., Alijanzadeh, D., Mazhari, S. A., Khademi, R., Kheirandish, A., & Naziri, M. (2024). Flavonoids against depression: a comprehensive review of literature. Frontiers in pharmacology, 15, 1411168. https://doi.org/10.3389/fphar.2024.1411168.

Alqurashi, G. K. (2022). The Impact of Chronic Unpredictable Mild Stress-Induced. MDPI Behavioral Sciences. https://www.mdpi.com/2076-328X/12/6/166.

Becker, M., Pinhasov, A., & Ornoy, A. (2021). Animal Models of Depression: What Can They Teach Us about the Human Disease?. Diagnostics (Basel, Switzerland), 11(1), 123. https://doi.org/10.3390/diagnostics11010123.

Belzung, C., Willner, P., & Philippot, P. (2015). Depression: From psychopathology to pathophysiology. Current Opinion in Neurobiology, 30, 24–30. https://doi.org/10.1016/j.conb.2014.08.013.

Boonruamkaew, P., Sukketsiri, W., Panichayupakaranant, P., Kaewnam, W., Tanasawet, S., Tipmanee, V., Hutamekalin, P., & Chonpathompikunlert, P. (2017). Apium graveolens extract influences mood and cognition in healthy mice. Journal of natural medicines, 71(3), 492–505. https://doi.org/10.1007/s11418-017-1077-6.

Brunton, L. L., Hilal-Dandan, R., & Knollmann, B. C. (2023). Goodman & Gilman’s the pharmacological basis of therapeutics (14th ed.). McGraw-Hill.

Can, A., Dao, D. T., Arad, M., Terrillion, C. E., Piantadosi, S. C., & Gould, T. D. (2012). The mouse forced swim test. Journal of Visualized Experiments, (59), e3638. https://doi.org/10.3791/3638.

Canenguez Benitez, J. S., Hernandez, T. E., Sundararajan, R., Sarwar, S., Arriaga, A. J., Khan, A. T., Benitez, G. A. (2022). Advantages and disadvantages of using St. John’s wort as a treatment for depression. Cureus, 14(9), e29468. https://doi.org/10.7759/cureus.29468.

Castrén, E., & Monteggia, L. M. (2021). Brain-Derived Neurotrophic Factor Signaling in Depression and Antidepressant Action. Biological Psychiatry, 90(2), 128–136. https://doi.org/https://doi.org/10.1016/j.biopsych.2021.05.008.

Chauhan, S., Tiwari, A., Verma, A., Padhan, P. K., Verma, S., & Gupta, P. C. (2024). Exploring the Potential of Saffron as a Therapeutic Agent in Depression Treatment: A Comparative Review. The Yale journal of biology and medicine, 97(3), 365–381. https://doi.org/10.59249/XURF4540.

Chen, X., Mao, R., Zou, Y., Yue, W., Dong, W., & Zhang, Y. (2025). Preliminary Exploration on the Effects of a Novel Antidepressant Formula Food in a Mouse Model. Foods, 14(9), 1640. https://doi.org/10.3390/foods14091640.

Commons, K. G., Cholanians, A. B., Babb, J. A., & Ehlinger, D. G. (2017). The rodent forced swim test measures stress-coping strategy, not depression-like behavior. ACS Chemical Neuroscience, 8(5), 955–960. https://doi.org/10.1021/acschemneuro.7b00042.

Cryan, J. F., & Holmes, A. (2015). The ascent of mouse: Advances in modelling human depression and anxiety. Nature Reviews Drug Discovery, 14(5), 317–336. https://doi.org/10.1038/nrd4505.

Cui, L., Li, S., Wang, S., Wu, X., Liu, Y., Yu, W., Wang, Y., Tang, Y., Xia, M., Li, B. Major depressive disorder: hypothesis, mechanism, prevention and treatment. Sig Transduct Target Ther 9, 30 (2024). https://doi.org/10.1038/s41392-024-01738-y.

Cui, Y., Ursuliak, Z., & Rupasinghe, H. P. V. (2025). Mechanistic insights into the antidepressant potential of plant-derived flavonoids: A preclinical review. Progress in Neuro-Psychopharmacology and Biological Psychiatry, 143, 111567. https://doi.org/https://doi.org/10.1016/j.pnpbp.2025.111567.

Fazilat, S., Tahmasbi, F., Mirzaei, M. R., Sanaie, S., Yousefi, Z., Asnaashari, S., Yaqoubi, S., Banagozar Mohammadi, A., & Araj-khodaei, M. (2024). A systematic review on the use of phytotherapy in managing clinical depression. BioImpacts, 15, 30532. https://doi.org/10.34172/bi.30532.

Frost, R., Zamri, A., Mathew, S., Salame, A., Bhanu, C., Bhamra, S. K., Bazo-Alvarez, J. C., Heinrich, M., & Walters, K. (2025). Understanding the research landscape of over-the-counter herbal products, dietary supplements, and medications evaluated for depressive symptoms in adults: a scoping review. Frontiers in Pharmacology, Volume 16-2025. https://doi.org/10.3389/fphar.2025.1609605.

Gençtürk, S., & Ünal, S. (2024). Rodent tests of depression and anxiety: Construct validity and translational relevance. Cognitive, Affective & Behavioral Neuroscience. https://doi.org/10.3758/s13415-024-01171-2.

Gómez-Patiño, M. B., Estrada-Reyes, R., Hernández-Mendoza, H. H., Suarez-Rojas, Á., & Arrieta-Baez, D. (2025). Antidepressant- and Anxiolytic-like Effects in Mice of Alkaloids from Aerial Parts of Argemone platyceras Link & Otto. Pharmaceuticals, 18(1), 49. https://doi.org/10.3390/ph18010049.

Kang, D., Dong, H., Shen, Y., Ou, J., & Zhao, J. (2023). The clinical application of Chinese herbal medication to depression: A narrative review. Frontiers in Public Health, 11, Article 1120683. https://doi.org/10.3389/fpubh.2023.1120683.

Kasper, S., Volz, H.-P., Möller, H.-J., Schläfke, S., Klement, S., Anghelescu, I.-G., & Seifritz, E. (2024). Lavender oil preparation Silexan is effective in mild-to-moderate major depression: A randomized, placebo- and reference-controlled trial. European Archives of Psychiatry and Clinical Neuroscience. Advance online publication. https://doi.org/10.1007/s00406-024-01783-2.

Kraeuter, A. K., Guest, P. C., & Sarnyai, Z. (2019). The forced swim test for depression-like behavior in rodents. Methods in Molecular Biology, 1916, 75–80. https://doi.org/10.1007/978-1-4939-8994-2_5.

Kusuma, I. W., Arung, E. T., Rosamah, E., & Kim, Y. U. (2018). Sedative activity of fractions of Apium graveolens leaves in mice. Pharmaceutical Biology, 56(1), 140–146. https://doi.org/10.1080/13880209.2018.1433185.

Lu, X. X., Tang, H., Li, X. H. (2025). Selecting an appropriate stress model of depression in rodents. World J Psychiatry, 15(12). https://doi.org/10.5498/wjp.v15.i12.113433.

Markov, D. D., & Novosadova, E. V. (2022). Chronic Unpredictable Mild Stress Model of Depression: Possible sources of poor reproducibility and latent variables. Biology, 11(11), Article 1621. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9687170/.

Mehrdad, S., Najmeh, A., Hossein, A., Zahra, M., Ali, A., Zahra, L.G. (2019). Phytochemical Study and Antidepressant Effect of Essential Oil of Apium graveolens L. Journal of Pharmaceutical Research International, 27(4), 1–10. https://doi.org/10.9734/jpri/2019/v27i430179.

Meyer, J. H., Ginovart, N., Boovariwala, A., Sagrati, S., Hussey, D., Garcia, A., Young, T., Praschak-Rieder, N., Wilson, A. A., & Houle, S. (2006). Elevated monoamine oxidase a levels in the brain: an explanation for the monoamine imbalance of major depression. Archives of general psychiatry, 63(11), 1209–1216. https://doi.org/10.1001/archpsyc.63.11.1209.

Mezadri, T. J., Batista, G. M., Portes, A. C., Marino-Neto, J., & Lino-de-Oliveira, C. (2011). Repeated rat-forced swim test: Reducing the number of animals to evaluate gradual effects of antidepressants. Journal of Neuroscience Methods, 195(2), 200–205. https://doi.org/https://doi.org/10.1016/j.jneumeth.2010.12.015.

Ministry of Health of the Republic of Indonesia. (2017). Indonesian herbal pharmacopoeia (2nd ed.). Jakarta: Directorate General of Pharmaceutical and Medical Devices.

Molendijk, M. L., & de Kloet, E. R. (2022). Forced swim stressor: Trends in usage and mechanistic consideration. The European journal of neuroscience, 55(9-10), 2813–2831. https://doi.org/10.1111/ejn.15139.

Momin, R. A., & Nair, M. G. (2002). Antioxidant, cyclooxygenase and topoisomerase inhibitory compounds from Apium graveolens Linn. seeds. Phytomedicine : international journal of phytotherapy and phytopharmacology, 9(4), 312–318. https://doi.org/10.1078/0944-7113-00131.

Nollet, M. (2021). Models of Depression: Unpredictable Chronic Mild Stress in Mice. Current Protocols, Wiley Periodicals LLC. https://pubmed.ncbi.nlm.nih.gov/34406704/.

Nurfahanum, R. (2022). Adverse effects and treatment resistance associated with long-term antidepressant use. Journal of Clinical Pharmacy Research, 5(1), 33–41.

Olasehinde, T. A., & Olaokun, O. O. (2024). The Beneficial Role of Apigenin against Cognitive and Neurobehavioural Dysfunction: A Systematic Review of Preclinical Investigations. Biomedicines, 12(1), 178. https://doi.org/10.3390/biomedicines12010178.

Olayinka, J. N., Akawa, O. B., Ogbu, E. K., Eduviere, A. T., Ozolua, R. I., & Soliman, M. (2023). Apigenin attenuates depressive-like behavior via modulating monoamine oxidase A enzyme activity in chronically stressed mice. Current research in pharmacology and drug discovery, 5, 100161. https://doi.org/10.1016/j.crphar.2023.100161.

Patel P, Saadabadi A. Monoamine Oxidase Inhibitors (MAOIs). In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2025 Jan. Available from: https://www.ncbi.nlm.nih.gov/books/NBK539848/.

Petit-Demoulière, B., Chenu, F., & Bourin, M. (2005). Forced swimming test in mice: A review of antidepressant activity. Methods and Findings in Experimental and Clinical Pharmacology, 27(8), 635–638. https://doi:10.1358/mfec.2005.27.8.955017.

Popović, M., Kaurinović, B., Trivić, S., Mimica-Dukić, N., & Bursać, M. (2006). Effect of celery (Apium graveolens) extracts on some biochemical parameters of oxidative stress in mice treated with carbon tetrachloride. Phytotherapy research : PTR, 20(7), 531–537. https://doi.org/10.1002/ptr.1871.

Remali, J., & Aizat, W. M. (2024). Medicinal plants and plant-based traditional medicine: Alternative treatments for depression and their potential mechanisms of action. Heliyon, 10(20), e38986. https://doi.org/10.1016/j.heliyon.2024.e38986.

Rosas-Sánchez, G. U., Germán-Ponciano, L. J., Guillen-Ruiz, G., Cueto-Escobedo, J., Limón-Vázquez, A. K., Rodríguez-Landa, J. F., & Soria-Fregozo, C. (2024). Neuroplasticity and Mechanisms of Action of Acute and Chronic Treatment with Antidepressants in Preclinical Studies. Biomedicines, 12(12), 2744. https://doi.org/10.3390/biomedicines12122744.

Sahin Z. (2023). Assessment of commonly used tests in experimental depression studies according to behavioral patterns of rodents. Medical review (2021), 3(6), 526–531. https://doi.org/10.1515/mr-2023-0018.

Sahin, Z., Solak, H., Koc, A., Ozen Koca, R., Ozkurkculer, A., Cakan, P., Solak Gormus, Z. I., Kutlu, S., & Kelestimur, H. (2019). Long-term metabolic cage housing increases anxiety/depression-related behaviours in adult male rats. Archives of physiology and biochemistry, 125(2), 122–127. https://doi.org/10.1080/13813455.2018.1441314.

Sánchez-Mateo, C. C., Bonkanka, C. X., Hernández-Pérez, M., & Rabanal, R. M. (2012). Evaluation of antidepressant activity of plant extracts in rodents. Journal of Ethnopharmacology, 142(1), 36–43. https://doi.org/10.1016/j.jep.2012.04.021.

Serefko, A., Lachowicz-Radulska, J., Szopa, A., Herbet, M., Czylkowska, A., Ignatiuk, K., Dołoto, A., Szewczyk, B., Wośko, S., Wróbel, A., Szponar, J., Wlaź, P., Skałecki, P., Wróbel, J., Słotwińska, W., & Poleszak, E. (2025). The Novel Imipramine–Magnesium Complex Exerts Antidepressant-like Activity in Mice Subjected to the Forced Swim Test and the Tail Suspension Test. Molecules, 30(3), 519. https://doi.org/10.3390/molecules30030519.

Slattery, D. A., & Cryan, J. F. (2017). Modelling depression in animals: At the interface of reward and stress pathways. Psychopharmacology, 234(9–10), 1451–1465. https://doi.org/10.1007/s00213-017-4550-3.

Stahl, S. M. (2023). Stahl’s essential psychopharmacology (5th ed.). Cambridge University Press.

Tan, S. H., Ahmad, R., & Ismail, Z. (2023). Phytochemical profiling and neuropharmacological potential of Apium graveolens. Journal of Herbal Medicine, 38, 100646. https://doi.org/10.1016/j.hermed.2023.100646.

Tan, T. Y. C., Lim, X. Y., Norahmad, N. A., Chanthira Kumar, H., Teh, B. P., Lai, N. M., & Syed Mohamed, A. F. (2023). Neurological Applications of Celery (Apium graveolens): A Scoping Review. Molecules (Basel, Switzerland), 28(15), 5824. https://doi.org/10.3390/molecules28155824.

Taylor, D. M., Barnes, T. R. E., & Young, A. H. (2021). The Maudsley prescribing guidelines in psychiatry (14th ed.). Wiley-Blackwell.

Tie, D., Wang, Y., Zhou, J., Zhang, Y., Ji, H., Yu, Y., Han, H., Xiang, Z., & Li, W. (2025). Baihe Dihuang Tang Exerts Antidepressant Effects via Modulation of MAOA-Mediated Serotonin Metabolism and Synaptic Plasticity. Pharmaceuticals, 18(12), 1786. https://doi.org/10.3390/ph18121786.

World Health Organization. (2017). Depression and other common mental disorders: Global health estimates. Geneva: World Health Organization. https://www.who.int/publications/i/item/depression-global-health-estimates.