Original Article

Anti-Toxoplasma Effect of Hydroalchohlic Extract of Terminalia chebula Retz in Cell Culture and Murine Model


Background: We examined anti-Toxoplasma effect of hydroalcoholic extract of Terminalia chebula Retz (T. chebula) in cell culture and murine model.

Methods: The study was conducted in Shahrekord University of Medical Sciences, Iran in 2017. Half maximal effective (concentration (EC50) of T. chebula extract and pyrimethamine was determined in infected Hela cells by using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) method. In the animal model, BALB/c mice were injected with tachyzoites (104) of T. RH strain intraperitoneally. 24h after the injection, the test groups were orally treated with 100, 200, 400 and 800 mg/kg of T. chebula extract for 7 days. The survival rate of the mice was determined and blood samples were collected to determine the amount of serum Malondialdehyde (MDA) and antioxidant capacity. Then peritoneal fluid of the mice was collected to count the number of tachyzoites and after necropsy, the pathologic changes, including the weight of liver, spleen and kidneys were investigated. The analysis of data was accomplished using SPSS.

Results: EC50 values were 94.7μg/mL and 290.50μg/mL for T. chebula and pyrimethamine respectively. In the animal model, the extract of T. chebula in concentration of 100 mg/kg showed the same anti-Toxoplasma effect as pyrimethamine. This concentration of the extract decreased number of intraperitoneal tachyzoites and increased the survival rate of the mice. This extract reduced the levels of serum MDA and tissue inflammation and increased serum antioxidant capacity.

Conclusion: Regarding the positive effect of extract, after more clinical trials in the animal model and standardization of the extract, it can be used as an alternative or complementary therapy for toxoplasmosis.

1. Dubey J, Lindsay D, Speer C. Structures of Toxoplasma gondii tachyzoites, bradyzoites, and sporozoites and biology and development of tissue cysts. Clin Microbiol Rev. 1998;11(2):267-299.
2. Duncanson P, Terry RS, Smith JE, et al. High levels of congenital transmission of Toxoplasma gondii in a commercial sheep flock. Int J Parasitol. 2001;31(14):1699-703.
3. Dubey JP. Strategies to reduce transmission of Toxoplasma gondii to animals and humans. Vet Parasitol. 1996;64(1-2):65-70.
4. Heukelbach J, Meyer-Cirkel V, Moura RCS, et al. Waterborne toxoplasmosis, northeastern Brazil. Emerg Infect Dis. 2007;13(2):287-289.
5. Dubey J, Jones J. Toxoplasma gondii infection in humans and animals in the United States. Int J Parasitol. 2008;38(11):1257-78.
6. Tenter AM, Heckeroth AR, Weiss LM. Toxoplasma gondii: from animals to humans. Int J Parasitol. 2000;30(12):1217-58.
7. Kurokawa H, Kato K, Iwanaga T, et al. Identification of Toxoplasma gondii cAMP dependent protein kinase and its role in the tachyzoite growth. PLoS One. 2011;6(7):e22492.
8. Goldstein EJ, Montoya JG, Remington JS. Management of Toxoplasma gondii infection during pregnancy. Clin Infect Dis. 2008;47(4):554-66.
9. Ryan P, Hurley SF, Johnson AM, et al. Tumours of the brain and presence of antibodies to Toxoplasma gondii. Int J Epidemiol. 1993;22(3):412-9.
10. Sukthana Y. Toxoplasmosis: beyond animals to humans. Trends Parasitol. 2006;22(3):137-42.
11. Fuller NJ, Bates CJ, Hayes RJ, et al. The effects of antimalarials and folate supplements on haematological indices and red cell folate levels in Gambian children. Ann Trop Paediatr. 1988;8(2):61-7.
12. Schmidt DR, Hogh B, Andersen O, et al. Treatment of infants with congenital toxoplasmosis: tolerability and plasma concentrations of sulfadiazine and pyrimethamine. Eur J Pediatr 2006;165(1):19-25.
13. Jiang J-H, Jin C-M, Kim Y-C, et al. Anti-toxoplasmosis effects of oleuropein isolated from Fraxinus rhychophylla. Bio and Pharma Bul. 2008;31(12):2273-6.
14. Rathinamoorthy R, Thilagavathi G. Terminalia chebula-review on pharmacological and biochemical studies. Int J Pharmtech Res. 2014;6(1):97-6.
15. Shinde SL, More S, Junne S, et al. The antifungal activity of five Terminalia species checked by paper disk method. Int J Pharma Res Dev. 2011;3:36-40.
16. Sohni YR, Bhatt RM. Activity of a crude extract formulation in experimental hepatic amoebiasis and in immunomodulation studies. J Ethnopharmacol. 1996;54(2-3):119-24.
17. Sahar E, Meshelhy MR, Kusumoto IT, et al. Inhibitory effects of egyptian folk medicines oh human immunodeficiency virus (HIV) reverse transcriptase. Chem Pharm Bull (Tokyo). 1995;43(4):641-8.
18. Cheng H-Y, Lin T-C, Yu K-H, et al. Antioxidant and free radical scavenging activities of Terminalia chebula. Biol Pharm Bull. 2003;26(9):1331-5.
19. Arora S, Kaur K, Kaur S. Indian medicinal plants as a reservoir of protective phytochemicals. Teratog Carcinog Mutagen. 2003; 1:295-300.
20. Reddy DB, Reddy T, Jyotsna G, et al. Chebulagic acid, a COX–LOX dual inhibitor isolated from the fruits of Terminalia chebula Retz., induces apoptosis in COLO-205 cell line. J ethnopharmacol. 2009;124(3):506-12.
21. Pinmai K, Hiriote W, Soonthornchareonnon N, et al. In vitro and in vivo antiplasmodial activity and cytotoxicity of water extracts of Phyllanthus emblica, Terminalia chebula, and Terminalia bellerica. J Med Assoc Thai. 2010; 7:S120-6.
22. Kavitha N, Noordin R, Chan K-L, et al . In vitro anti-Toxoplasma gondii activity of root extract/fractions of Eurycoma longifolia Jack. BMC Complement Altern Med. 2012;12:91.
23. Malekzadeh F, Ehsanifar H, Shahamat M, et al. Antibacterial activity of black myrobalan (Terminalia chebula Retz) against Helicobacter pylori. Int J Antimicrob Agents. 2001;18(1):85-8.
24. Rabbani M, Sajjadi S, Jalali A. Hydroalcohol extract and fractions of Stachys lavandulifolia vahl: effects on spontaneous motor activity and elevated plus‐maze behaviour. PhytotherRes. 2005;19(10):854-8.
25. Choi K-M, Gang J, Yun J. Anti-Toxoplasma gondii RH strain activity of herbal extracts used in traditional medicine. Int J Antimicrob Agents. 2008;32(4):360-2.
26. Yang X, Huang B, Chen J, et al. In vitro effects of aqueous extracts of Astragalus membranaceus and Scutellaria baicalensis GEORGI on Toxoplasma gondii. Parasitol Res. 2012;110(6):2221-7.
27. Choi W, Jiang M, Chu J. Antiparasitic effects of Zingiber officinale (Ginger) extract against Toxoplasma gondii. Journal of Applied Biomedicine. 2013;11(1):15-26.
28. de Oliveira TC, Silva DAO, Rostkowska C, et al. Toxoplasma gondii: effects of Artemisia annua L. on susceptibility to infection in experimental models in vitro and in vivo. Exp Parasitol. 2009;122(3):233-41.
29. Romand S, Pudney M, Derouin F. In vitro and in vivo activities of the hydroxy naphthoquinone atovaquone alone or combined with pyrimethamine, sulfadiazine, clarithromycin, or minocycline against Toxoplasma gondii. Antimicrob Agents Chemother. 1993;37(11):2371-8.
30. Khoshzaban F, Ghafarifar F, Sharafi M, et al. Effects of Peganum harmala on acute toxoplasmosis in mice. Daneshvar Medicine. 2008;15(75):27-36.
31. Irandoost H, Yazdinezhad A, Fazaeli A. The Effects of Ethanol Extract of Propolis on Acute Toxoplasmosis in Mouse Model. J Zanjan Univ Med Sci. 2015; 23(100): 86-94.
32. Khoshzaban F, Ghafarifar F, Sharafi M, et al. The effect of garlic extract on acute toxoplasmosis in mice. Iranian Journal Of Medicinal And Aromatic Plants. 2007;23(3):295-306.
33. Hagege D, Nouvelot A, Boucaud J, et al. Malondialdehyde titration with thiobarbiturate in plant extracts: avoidance of pigment interference. Phytochem Anal. 1990;1(2):86-89.
34. Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Analy Biochem. 1996;239(1):70-76.
35. Luque‐Sierra A, Alvarez‐Amor L, Kleemann R, et al. Extra‐Virgin Olive Oil with Natural Phenolic Content Exerts an Anti‐Inflammatory Effect in Adipose Tissue and Attenuates the Severity of Atherosclerotic Lesions in Leiden Mice. Mol Nutr Food Res. 2018;62(13):1800295.
36. Mitchell SM, Zajac AM, Davis WL, et al. Efficacy of ponazuril in vitro and in preventing and treating Toxoplasma gondii infections in mice. J Parasitol. 2004;90(3):639-42.
37. Montazeri M, Sharif M, Sarvi S, et al. A systematic review of in vitro and in vivo activities of anti-Toxoplasma drugs and compounds (2006–2016). Front Microbiol. 2017;8:25.
38. Jafari M, Naeini KM, Lorigooini Z, et al. Oral acute and sub-acute toxic effects of hydroalcoholic Terminalia chebula Retz and Achillea wilhelmsii extracts in BALB/c mice. BioMedicine. 2019;9(4):25.
39. Rafieian-Kopaei M, Baradaran A, Rafieian M. Oxidative stress and the paradoxical effects of antioxidants. Int J Res Med Sci. 2013;18(7):629.
40. Karaman U, Celik T, Kiran TR, et al. Malondialdehyde, glutathione, and nitric oxide levels in Toxoplasma gondii seropositive patients. Korean J Parasitol. 2008;46(4):293-295.
41. Bag A, Bhattacharyya SK, Chattopadhyay RR. The development of Terminalia chebula Retz.(Combretaceae) in clinical research. Asian Pac J Trop Biomed. 2013;3(3):244-252.
IssueVol 16 No 4 (2021) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v16i4.7876
Toxoplasma gondii Terminalia chebula Retz Murine Tetrazolium salts (MTT) Hela cells

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Jafari M, Lorigooini Z, Kheiri S, Manouchehri Naeini K. Anti-Toxoplasma Effect of Hydroalchohlic Extract of Terminalia chebula Retz in Cell Culture and Murine Model. Iran J Parasitol. 2021;16(4):631-640.