Leishmanicidal Effects of Allium giganteum Saponin-Rich Fraction on Leishmania major

Document Type : Original paper


1 Department of Parasitology & Mycology, School of Medicine, Isfahan University of Medical Sciences & Health Services, Isfahan, Iran.

2 Department of Pharmacognosy, School of pharmacy, Isfahan University of Medical Sciences & Health Services, Isfahan, Iran.

3 Research Committee, School of Pharmacy, Isfahan University of Medical Science & Health Services, Isfahan, Iran.

4 Department of Parasitology and Mycology, School of Medicine, Isfahan University of Medical Sciences & Health Services, Isfahan, Iran.


Background and objectives: Leishmaniasis is caused by the genus of Leishmania and is one of the important health problems worldwide. Serious side effects, the lack of effective vaccines and the emergence of drug resistance are the major weak points of leishmaniasis treatment. The purpose of this study was to evaluate leishmanicidal effects of Allium giganteum saponin rich fraction, natural compounds with history of antimicrobial properties, on promastigotes and axenic amastigotes of L. major and macrophages cell line J774. Methods: The chloroform-methanol (9:1) extract of the flowers was fractionated by MPLC using an RP-18 column. The saponin-rich fraction was detected by TLC and H-NMR analyses and evaluated for leishmanicidal activity on L. major and macrophages cell line J774 using MTT assay at 24, 48 and 72 h of incubation. Results: At concentrations of 75, 100 and 150 μg/mL, over the time of 24 to 72 h, a significant decrease in the live parasite's rate was observed (p <0.05). At 200 μg/mL concentration, all parasites were killed and maximum leishmanicidal effect was observed. The IC50s for promastigotes and axenic amastigotes were 90.01 ± 13.42 μg/mL and 29.76±17.91 μg/mL, respectively; the value for the J774 macrophage cell line was 33.17±4 μg/mL. Conclusion: The results of this study showed the significant leishmanicidal effect of saponin rich fraction from Allium giganteum on promastigote and axenic amastigote of L. major and macrophage cell line in vitro. Complementary in vivo studies for evaluating the effects of the fraction on leishmaniasis in BALB/c mice is recommended.


Main Subjects

[1] Eskandarian A, Mohaghegh M, Yousefi H. In vitro antileishmanial activity of Falcaria                   vulgaris fractions on Leishmania major.  Jundishpur J Nat Pharm Prod. 2017; Article ID 63754.
[2] Shirzadi M, Esfahania S, Mohebalia M, Ershadia M, Gharachorlo F, Razavia M, Postigo JAR. Epidemiological status of leishmaniasis in the Islamic Republic of Iran, 1983-2012. East Mediterr Health J. 2015; 21(10): 736-742.
[3] Sundar S, Goyal N. Molecular mechanisms of antimony resistance in Leishmania. J Med Microbiol. 2007; 56(2): 143-153.
[4] Hefnawy A, Berg M, Dujardin JC, De Muylder G. Exploiting knowledge on Leishmania drug resistance to support the quest for new drugs. Trends Parasitol. 2017; 33(3): 162-174.
[5] Ait-Oudhia K, Gazanion E, Oury B, Vergnes B, Sereno D. The fitness of antimony-resistant Leishmania parasites: lessons from the field. Trends Parasitol. 2011; 27(4): 141-142.
[6] Khatami A, Firooz A, Gorouhi F, Dowlati Y. Treatment of acute old world cutaneous leishmaniasis: a systematic review of the randomized controlled trials. J Am Acad Dermatol. 2007; 57(2): 1-29.
[7] Hadighi R, Mohebali M, Boucher P, Hajjaran H, Khamesipour A, Ouellette M. Unresponsiveness to glucantime treatment in Iranian cutaneous leishmaniasis due to drug-resistant Leishmania tropica parasites. PLoS Med. 2006; Article ID 16605301.
[8] Mishra BB, Kale RR, Singh RK, Tiwari VK. Alkaloids: future prospective to combat leishmaniasis. Fitoterapia. 2009; 80(2): 81-90.
[9] Santos WR, de Lima VM, de Souza EP, Bernardo RR, Palatnik M, de Sousa CBP. Saponins, IL12 and BCG adjuvant in the FML-vaccine formulation against murine visceral leishmaniasis. Vaccine. 2002; 21(1-2): 30-43.
[10] Maes L, Berghe DV, Germonprez N, Quirijnen L, Cos P, De Kimpe N, Puyvelde LV. In vitro and in vivo activities of a triterpenoid saponin extract (PX-6518) from the plant Maesa balansae against visceral Leishmania species. Antimicrob Agents Chemother. 2004; 48(1): 130-136.
[11] Ahmad I, Beg AZ. Antimicrobial and phytochemical studies on 45 Indian medicinal plants against multi-drug resistant human pathogens. J Ethnopharmacol. 2001; 74(2): 113-123.
[12] Hostettmann K, Marston A. Saponins. 1st ed. Cambridge: Cambridge University Press, 1995.
[13] Tava A, Avato P. Chemical and biological activity of triterpene saponins from Medicago      species. Nat Prod Commun. 2006: 1(12): 1159-1180.
[14] Tabassum Khan N. Therapeutic properties of saponins. Int Invent Sci J. 2019; 3(1): 409-411.
[15] Dutta A, Ghoshal A, Mandal D, Mondal N, Banerjee S, Sahu N, Mandal Ch. Racemoside A, an anti-leishmanial, water-soluble, natural steroidal saponin, induces programmed cell death in Leishmania donovani. J Med Microbiol. 2007; 56(9): 1196-1204.
[16] Arote S, Dahikar S, Yeole P. Phytochemical screening and antibacterial properties of leaves of Pongamia pinnata Linn. (Fabaceae) from India. African J Biotech. 2009; 8(22): 6393-6396.
[17] Cioaca C, Margineanu C, Cucu V. The saponins of Hedera helix with antibacterial activity. Die Pharmazie. 1978; 33(9): 609-610.
[18] Barile E, Bonanomi G, Antignani V, Zolfaghari B, Sajjadi SE, Scala F, Lanzotti V. Saponins from Allium minutiflorum with antifungal activity. Phytochemistry. 2007; 68(5): 596-603.
[19] Baghalian K, Naghavi MR, Ziai SA, Badi HN. Post-planting evaluation of morphological characters and allicin content in Iranian garlic (Allium sativum L.) ecotypes. Sci Hortic. 2006; 107(4): 405-410.
[20] Azwanida N. A review on the extraction methods use in medicinal plants, principle, strength and limitation. Med Aromat Plants. 2015; 4(3): 1-6.
[21] Teixeira M, de Jesus Santos R, Sampaio R, Pontes-de-Carvalho L, dos-Santos WL. A simple and reproducible method to obtain large numbers of axenic amastigotes of different Leishmania species. Parasitol Res. 2002; 88(11): 963-968.
[22] Young FM, Phungtamdet W, Sanderson BJS. Modification of MTT assay conditions to examinethe cytotoxic effects of amitraz on the humanlymphoblastoid cell line, WIL2NS. Toxicol In Vitro. 2005; 19(8): 1051-1059. 
[23] Sadeghi M, Zolfaghari B, Troiano R, Lanzotti V. 3-Keto umbilicagenin A and B, new sapogenins from Allium umbilicatum Boiss. Fitoterapia. 2015; 102(1): 198-202.
[24] Rezaee F, Zolfaghari B, Dinani MS. Isolation of dioscin-related steroidal saponin from the bulbs of Allium paradoxum L. with leishmanicidal activity. Res Pharm Sci. 2018; 13(5): 469-475.
[25] Norouzinezhad F, Ghaffari F, Norouzinejad A, Kaveh F, Gouya MM. Cutaneous leishmaniasis in Iran: results from an epidemiological study in urban and rural provinces. Asian Pacific J Tropic Biomed. 2016; 6(7): 614-619.
[26] Sajjadi SE, Eskandarian AA, Shokoohinia Y, Yousefi HA, Mansourian M, Asgarian-Nasab H, Mohseni N. Antileishmanial activity of prenylated coumarins isolated from Ferulago angulata and Prangos asperula. Res Pharm Sci. 2016; 11(4): 324-331.
[27] Mojarrab M, Shiravand A, Delazar A, Heshmati Afshar F. Evaluation of in vitro antimalarial activity of different extracts of Artemisia aucheri Boiss. and A. armeniaca Lam. and fractions of the most potent extracts. Sci World J. 2014; Article ID 825370.
[28] Gharirvand Eskandari E, Doudi M. Investigation of antileishmanial effect of alcoholic extract and essential oil of medicinal plant leaf black alfalfa (Medicago lupulina), on the number of clinical isolates of Leishmania major promastigotes in vitro. J Shahid Sadoughi Univ Med Sci. 2016; 24(2): 174-184.
[29] Ghazanfari T, Hassan ZM, Khamesipour A. Enhancement of peritoneal macrophage phagocytic activity against Leishmania major by garlic (Allium sativum) treatment. J Ethnopharmacol. 2006; 103(3): 333-337.
[30] Hejazi SH, Makvandi S, Abedi S, Soleimanifard S. The study of the inhibitory effects of aqueous and ethanolic extracts of Ziziphora tenuior L. on the proliferation of Leishmania major amastigotes. Iran J Dermatol. 2018; 21(3): 71-76.
[31] Emami SA, Rabe SZT, Ahi A, Mahmoudi M. Inhibitory activity of eleven Artemisia species from Iran against Leishmania major parasites. Iran J Basic Med Sci. 2012; 15(2): 807-811.
[32] Barati M, Sharifi L, Sharififar F. Antileishmanial activity of Artemisia aucheri, Ferula asafoetida and Gossypium hirsutum extracts on Leishmania major promastigotes in vitro. Ann Mill Health Sci Res. 2010; 8(31): 166-172.
[33] Rees L, Minney S, Plummer N, Slater J, Skyrme D. A quantitative assessment of the antimicrobial activity of garlic (Allium sativum). World J Microbiol Biotech. 1993; 9(3): 303-307.
[34] Khademvatan S, Saki J, Gharavi MJ, Rahim F. Allium sativum extract induces apoptosis in Leishmania major (MRHO/IR/75/ER) promastigotes. J Med Plants Res. 2011; 5(16): 3725-3732.
[35] Gamboa‐León M, Aranda‐González I, Mut‐Martín M, García‐Miss M, Dumonteil E. In vivo and in vitro control of Leishmania mexicana due to garlic‐induced NO production. Scand J Immunol. 2007; 66(5): 508-514.
[36] Ghazanfari T, Hassan Z, Ebtekar M, Ahmadiani A, Naderi G, Azar A. Garlic induces a shift in cytokine pattern in Leishmania major‐infected Balb/c mice. Scand J Immunol. 2000; 52(5): 491-495.
[37] Kuroda M, Mimaki Y, Kameyama A, Sashida Y, Nikaido T. Steroidal saponins from Allium chinense and their inhibitory activities on cyclic AMP phosphodiesterase and Na+ K+ ATPase. Phytochemistry. 1995; 40(4): 1071-1076.
[38] Patil P, Patil J, Mahale J, Patel J, Surana S. Evaluation of antiulcer activity of the terpenoid fraction from the leaves of Thespesia populnea (L.) (Malvaceae) in albino rats. Res J Pharm Biol Chem Sci. 2010; 1(4): 495-513.