Original Article

Cytotoxicity and Anti-Plasmodium berghei Activity of Emodin Loaded Nanoemulsion


Background: Malaria parasites cause a tremendous burden of disease in both the tropics and subtropics areas. Growing of drugs resistance in parasites is one of the most threats to malaria control. The aim of study was to investigate the anti-malarial activity of nano-emodin isolated from Rhamnus cathartica on Plasmodium berghei in mice to evaluate parasites inhibition rate using in-vivo test.

Methods: The study was conducted in the School of Public Health, Tehran University of Medical Sciences, during 2020. Nano- emodin particles were prepared from Rhamnus cathartica, and confirmed by Zeta Potential Analyzer, DLS and electron microscopy techniques. Mice were infected with P. berghei and treated by emodin nanoparticles. Parasitemia was evaluated in each group in comparison with control group. Toxicity test was done using twice the highest concentration of emodin extract on a separate group of mice and ED50 was calculated.

Results: Emodin extract was significantly effective in all concentrations on D4 (P<0.05). The most effective on parasitemia was observed in 400 mg/kg of Liquid Nano-emodin and solid (non-Nano) emodin. ED50 for emodin extract was determined 220 mg/kg. Toxicity test showed no toxic effect on the subjects.

Conclusion: The emodin extract is safe, lack of side effects. So, it can be used for more and longer period of time and in higher doses. Emodin extract, either in form of liquid and nanoparticle or in a solid form, has the same therapeutic effect on P. berghei in infected Balb/c mice.

1. Sintasath DM, Ghebremeskel T, Lynch M, et al. Malaria prevalence and associated risk factors in Eritrea. Am J Trop Med Hyg. 2005; 1;72(6):682-7.
2. Mairet-Khedim M, Leang R, Marmai C, et al. Clinical and in-vitro resistance of Plasmodium falciparum to artesunate-amodiaquine in Cambodia. Clin Infect Dis. 2021;73(3): 406-413.
3. Hanafi-Bojd AA, Vatandoost H, Jafari R. Susceptibility status of Anopheles dthali and An. fluviatilis to commonly used larvicides in an endemic focus of malaria, southern Iran. J Vector Borne Dis. 2006;43(1):34.
4. Willcox ML, Bodeker G. Traditional herbal medicines for malaria. BMJ.2004; 329(7475), 1156-1159.
5. Edwin GT, Korsik M, Todd MH. The past, present and future of anti-malarial medicines. Malar J. 2019; 18(1):1-21.
6. Burrows JN, Duparc S, Gutteridge WE, et al. New developments in anti-malarial target candidate and product profiles. Malar J. 2017;16(1):1-29.
7. Phanouvong S, Raymond C, Krech L, et al.The quality of anti-malarial medicines in western Cambodia: a case study along the Thai-Cambodian border. Southeast Asian J Trop Med Public Health.2013;44(3):349-62.
8. Panda S, Rout JR, Pati P, et al. Anti-malarial activity of Artemisia nilagirica against Plasmodium falciparum.2018; J Parasit Dis. 2018;42(1):22-7.
9. Izhaki I. Emodin–a secondary metabolite with multiple ecological functions in higher plants. New Phytol.2002;155(2):205-17.
10. Pourhajibagher M, Rahimi-Esboei B, Ahmadi H, et al. The anti-biofilm capability of nano-emodin-mediated sonodynamic therapy on multi-species biofilms produced by burn wound bacterial strains. Photodiagnosis Photodyn Ther. 2021;34:102288.
11. Chabra A, Rahimi-Esboei B, Habibi E, et al. Effects of some natural products from fungal and herbal sources on Giardia-lamblia in vivo. Parasitology. 2019;146(9):1188-98.
12. Wang J, Zhang Y, Zhu Q, et al. Data on the radioprotective effect of emodin in vivo and vitro via inhibition of apoptosis and modulation of p53. Data Brief. 2017;11:290-5.
13. Liu X, Shan K, Shao X, et al. Nanotoxic effects of silver nanoparticles on normal HEK-293 cells in comparison to cancerous HeLa cell line. Int J Nanomedicine.2021;16:753.
14. Rashidzadeh H, Tabatabaei Rezaei SJ, Adyani SM, et al. Recent advances in targeting malaria with nano-technology-based drug carriers. Pharm Dev Technol. 2021;26(8),807-823.
15. Dong X, Zeng Y, Liu Y, et al. Aloe‐emodin: a review of its pharmacology, toxicity, and pharmacokinetics. Phytother Res. 2020;34(2):270-81.
16. Xia S, Ni Y, Zhou Q, et al. Emodin attenuates severe acute pancreatitis via antioxidant and anti-inflammatory activity. Inflammation. 2019;42(6):2129-38.
17. Pourhajibagher M, Hodjat M, Bahador A. Sonodynamic excitation of nanomicelle curcumin for eradication of Streptococcus mutans under sonodynamic anti-microbial chemotherapy: Enhanced anti-caries activity of nanomicelle curcumin. Photodiagnosis Photodyn Ther. 202;30:101780.
18. Wolfensohn S, Lloyd M. Handbook of laboratory animal management and welfare. John Wiley & Sons. 2008; pages21-35.
19. Jongwutiwes S, Buppan P, Kosuvin R, et al. Plasmodium knowlesi malaria in humans and macaques, Thailand Emerg Infect Dis. 2011;17(10):1799.
20. Mazhari N, Nateghpour M, Heydarian P, et al. In-vivo Anti-Malarial Activity of Heracleum persicum Fruit Extract, in Combination with Chloroquine against Chloroquine–Sensitive Strain of Plasmodium berghei. Iran J Public Health. 2018;47(6), 868-74.
21. Frita R, Carapau D, Mota MM, et al. In vivo hemozoin kinetics after clearance of Plasmodium berghei infection in mice. Malar Res Treat. 2012, 2012:373086.
22. Legesse M, Erko B, Balcha F. Increased parasitaemia and delayed parasite clearance in Schistosoma mansoni and Plasmodium berghei co-infected mice. Acta Trop. 2004; 91(2):161-6.
23. Ken-Ezihuo SU, Itatat SS, Bartimaeus EAS. Comparative Study of Two Different Rapid Diagnostic Tests with Microscopy Method for Malaria Parasite Detection. J Adv Med. 2019; 1-8.
24. Peters W, Robinson B. The chemotherapy of rodent malaria. XLVII. Studies on pyronaridine and other Mannich base antimalarials. Ann Trop Med Parasitol.1992; 86(5):455-65.
25. Evans BC, Nelson CE, Shann SY, et al. Ex-vivo red blood cell hemolysis assay for the evaluation of pH-responsive endosomolytic agents for cytosolic delivery of bio-macromolecular drugs. J Vis Exp.2013; (73):e50166.
26. Fievet CJ, Gigandet MP, Ansel HC. Hemolysis of erythrocytes by primary pharmacologic agents. Am J Hosp Pharm.1976;28(12):961-6.
27. Jensen JB, Trager W. Plasmodium falciparum in culture: use of outdated erythrocytes and description of the candle jar method. J Parasitol. 1977; 883-6.
28. Karbalaei Pazoki Z, Nateghpour M, Maghsood A, et al. Comparison between the effects of ethanolic extract of Artemisia annua and chloroquine on Plasmodium berghei in white mice. Sci J Kurd Univ Med Sci. 2014;19(2):9-20.
29. Batista MN, Braga ACS, Campos GRF, et al. Natural products isolated from oriental medicinal herbs inactivate Zika virus. Viruses. 2019;11(1):49.
30. Nateghpour M, Farivar L, Souri E, et al. The effect of Otostegia persica in combination with chloroquine on chloroquine-sensitive and chloroquine-resistant strains of Plasmodium berghei using in-vivo fixed ratios method. Iran J Pharm Res. 2012;11(2):583.
31. Heydarian P, Nateghpour M, Mazhari N, et al. Evaluation of Effectiveness of Ethanolic-Extract of Curcuma longa, discretely and in Combination with Chloroquine against Chloroquine-Sensitive Strain of Plasmodium berghei. Herb Med J. 2019;3(4):133-8.
IssueVol 17 No 3 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v17i3.10624
Malaria Plasmodium berghei Nano emodin Rhamnus cathartica In-vivo test

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
Bayat F, Motevalli Haghi A, Nateghpour M, Rahimi-Esboei B, Rahimi Foroushani A, Amani A, Farivar L, Sayyad Talaee Z, Faryabi A. Cytotoxicity and Anti-Plasmodium berghei Activity of Emodin Loaded Nanoemulsion. Iran J Parasitol. 2022;17(3):339-348.