Abstract
The severe toxicity, exorbitant cost and the emerging resistance of Leishmania spp. against most of the currently used drugs led to the urgent need for exploiting our traditional Ayurvedic knowledge to treat visceral leishmaniasis. The aim of this study was to evaluate the in vitro anti-leishmanial activity of various extracts from ten traditionally used Indian medicinal plants. The methanolic extract from only two plants, Withania somnifera Dunal (ashwagandha) and Allium sativum Linn. (garlic), showed appreciable activity against Leishmania donovani. Further active compounds from these two plants were isolated and purified based on bioactivity-guided fractionation. HPLC-purified fraction A6 of ashwagandha and G3 of garlic showed consistently high activity with 50% inhibitory concentration (IC50) of 12.5 ± 4 and 18.6 ± 3 μg/ml against promastigotes whereas IC50 of 9.5 ± 3 and 13.5 ± 2 μg/ml against amastigote form, respectively. The fraction A6 of ashwagandha was identified as withaferin A while fraction G3 of garlic is yet to be identified, and the work is in progress. Cytotoxic effects of the promising fractions and compounds were further evaluated in the murine macrophage (J774G8) model and were found to be safe. These compounds showed negligible cytotoxicity against J774G8 macrophages. The results indicate that fraction A6 of ashwagandha and fraction G3 of garlic might be potential sources of new anti-leishmanial compounds. The in vivo efficacy study and further optimization of these active compounds are in progress.
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References
Augusti KT (1996) Therapeutic values of onion (Allium cepa L.) and garlic (Allium sativum L.). Indian J Exp Biol 34:634–640
Ballal M, Srujan D, Bhat KK, Shirwaikar A, Shivananda PG (2001) Antibacterial activity of Holarrhena antidysenterica (Kurchi) against the enteric pathogens. Indian J Pharmacol 33:392–393
Birdsall TC, Kelly GS (1997) Berberine: therapeutic potential of an alkaloid found in several medicinal plants. Altern Med Rev 2:94–103
Block E, Ahmad S (1984) (E,Z)-Ajoene: a potent antithrombotic agent from garlic. J Am Chem Soc 106:8295–8296
Croft SL, Coombs GH (2003) Leishmaniasis—current chemotherapy and recent advances in the search for novel drugs. Trends Parasitol 19:502–508
Cui L, Miao J, Liwang C (2007) Cytotoxic effect of curcumin on malaria parasite Plasmodium falciparum: inhibition of histone acetylation and generation of reactive oxygen species. Antimicrob Agents Chemother 51:488–494
Deshpande RG, Khan MB, Bhat DA, Navalkar RG (1993) Inhibition of Mycobacterium avium complex isolates from AIDS patients by garlic (Altium sativum). Antimicrob Agents Chemother 32:623–626
Desjeux P (2004) Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27:305–318
Edzard E (1998) Harmless herbs? A review of the recent literature. Am J Med 104:170–178
Gallwitz H, Bonse S, Martinez-Cruz A, Schlichting I, Schumacher K, Krauth-Siegel RL (1999) Ajoene is an inhibitor and subversive substrate of human glutathione reductase and Trypanosoma cruzi trypanothione reductase: crystallographic, kinetic, and spectroscopic studies. J Med Chem 42:364–372
Ghazanfari T, Hassan ZM, Ebtekar M, Ahmadiani A, Naderi G, Azar A (2000) Garlic induces a shift in cytokine pattern in Leishmania major-infected Balb/c mice. Scand J Immunol 52:491–495
Ghazanfari T, Hassan ZM, Khamesipour A (2006) Enhancement of peritoneal macrophage phagocytic activity against Leishmania major by garlic (Allium sativum) treatment. J Ethnopharmacol 103:333–337
Ghosh M (2009) Purification of a lectin-like antifungal protein from the medicinal herb, Withania somnifera. Fitoterapia 80:91–95
Hoet S, Opperdoes F, Brun R, Adjakidje V, Quetin-Leclercq J (2004) In vitro antitrypanosomal activity of ethnopharmacologically selected Beninese plants. J Ethnopharmacol 91:37–42
Kumar D, Mishra SK, Tandan SK, Tripathi HC (1995) Possible mechanism of antihelminthic action of palasonin on Ascaridia galli. Indian J Pharmacol 27:161–166
Owais M, Sharad KS, Shehbaz A, Saleemuddin M (2005) Antibacterial efficacy of Withania somnifera (ashwagandha) an indigenous medicinal plant against experimental murine salmonellosis. Phytomedicine 12:229–235
Patricia S, Samanta PA, Marcia SCM, Frederico OP, Andre GT (2007) Isolation of anti-leishmanial sterol from the fruits of Cassia fistula using bioguided fractionation. Phytother Res 21:644–647
Politi M, Alvaro-Blanco J, Groves P, Prieto A, Antonio Leal J, Javier Canada F, Jimenez-Barbero J (2006) Screening of garlic water extract for binding activity with cholera toxin B pentamer by NMR spectroscopy—an old remedy giving a new surprise. European J Org Chem 9:2067–2073
Rajani M, Gupta SK, Singh N, Mathur S, Kochupillai V, Velpandian T (2006) Evaluation of the effect of Withania somnifera root extracts on cell cycle and angiogenesis. J Ethnopharmacol 105:336–341
Rani P, Khullar N (2004) Antimicrobial evaluation of some medicinal plants for their anti-enteric potential against multi-drug resistant Salmonella typhi. Phytother Res 18:670–673
Rates SMK (2001) Plants as source of drugs. Toxicon 39:603–613
Redhu NS, Dey A, Balooni V, Singh S (2006) Leishmania–HIV co-infection: an emerging problem in India. AIDS 20:1213–1215
Schlein Y, Jacobson RL (1994) Mortality of Leishmania major in Phlebotomus papatasi caused by plant feeding of the sand flies. Am J Trop Med Hyg 50:20–27
Selma RP, Shirley SM, Figueiredo MR, Kaplan MAC (2003) Plumbaginales: a pharmacological approach. Floresta Ambient 10:98–105
Sen N, Banerjee B, Das BB, Ganguly A, Sen T, Pramanik S, Mukhopadhyay S, Majumder HK (2007) Apoptosis is induced in leishmanial cells by a novel protein kinase inhibitor withaferin A and is facilitated by apoptotic topoisomerase I–DNA complex. Cell Death Differ 14:358–367
Singh S, Sivakumar R (2004) Challenges and new discoveries in the treatment of leishmaniasis. J Infect Chemother 10:307–315
Sivakumar R, Sharma P, Singh S (2006) Cloning, expression and purification of a novel recombinant antigen from Leishmania donovani. Protein Expr Purif 46:156–165
Tada H, Shiho O, Kuroshima K, Koyama M, Tsukamoto K (1986) An improved colorimetric assay for interleukin 2. J Immunol Methods 93:157–165
Takahashi T, Kokubo R, Sakaino M (2004) Antimicrobial activities of eucalyptus leaf extracts and flavonoids from Eucalyptus maculata. Lett Appl Microbiol 39:60–64
Tiuman TS, Ueda-Nakamura T, Cortez DAG, Dias Filho BP, Morgado-Díaz JA, Souza W, Nakamura CV (2005) Anti-leishmanial activity of parthenolide, a sesquiterpene lactona isolated from Tanacetum parthenium. Antimicrob Agents Chemother 49:176–182
Urbina JA, Marchan E, Lazardi K, Visbal G, Apitz-Castro R, Gil F, Aguirre T, Piras MM, Piras R (1993) Inhibition of phosphatidylcholine biosynthesis and cell proliferation in Trypanosoma cruzi by ajoene, an antiplatelet compound isolated from garlic. Biochem Pharmacol 45:2381–2387
Ushimaru PI, Nogueira da Silva MT, Claudio Di Stasi L, Barbosa L, Fernandes Junior A (2007) Antibacterial activity of medicinal plant extracts. Braz J Microbiol 38:717–719
Venugopal PV, Venugopal TV (1994) Antidermatophytic activity of Neem (Azadirachta indica) leaves in vitro. Indian J Pharmacol 26:141–143
Acknowledgments
This study was supported by a grant from Central Council for Research in Unani Medicine, Department of AYUSH, Ministry of Health and F.W., Government of India, to SS. The financial assistance in the form of research fellowship from the Indian Council of Medical Research to US, New Delhi, is also acknowledged.
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Sharma, U., Velpandian, T., Sharma, P. et al. Evaluation of anti-leishmanial activity of selected Indian plants known to have antimicrobial properties. Parasitol Res 105, 1287–1293 (2009). https://doi.org/10.1007/s00436-009-1554-2
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DOI: https://doi.org/10.1007/s00436-009-1554-2