Holothuria leucospilota Extract Induces Apoptosis in Leishmania major Promastigotes
AbstractBackground: The present study aimed to survey antileishmanial activity of methanolic Holothuria leucospilota extract against Leishmania major promastigotes in vitro.Methods: Promastigotes were cultured in RPMI 1640 and after reaching the stationary phase, the study was conducted with different concentrations of the extract. Afterwards, MTT colorimetric assay for the obtaining of 50% inhibitory concentration (IC50) was utilized. Furthermore, in order to determine the possible induction of apoptosis in L. major promastigotes, flow cytometry and DNA fragmentation methods were employed using annexin-V FLUOS staining kit and DNA ladder kit, respectively.Results: The IC50 value of H. leucospilota extract at three time points of 24, 48, and 72 h was estimated 2000, 300 and 85 µg/ml, respectively. In addition, the extract revealed a dose and time-dependent antileishmanial activity. Furthermore, various characteristics of apoptosis appeared after L. major promastigotes treatment, which included cell shrinkage, formation of apoptotic bodies, blebbing of the cell membrane, and externalization of phosphatidylserine, although no laddering pattern was observed.Conclusion: The methanolic extract of H. leucospilota possesses lethal effect on L. major promastigotes and induces the apoptosis in parasites. Further studies are required to address the apoptosis mechanism in vivo.
Desjeux P. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis. 2004;27(5):305-18.
Akhoundi M, Kuhls K, Cannet A, Votypka J, Marty P, Delaunay P, et al. A historical overview of the classification, evolution, and dispersion of Leishmania parasites and sandflies. PLoS Negl Trop Dis. 2016;10(3):e0004349.
Alvar J, Velez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 2012;7(5):e35671.
Karimi A, Hanafi-Bojd AA, Yaghoobi-Ershadi MR, Akhavan AA, Ghezelbash Z. Spatial and temporal distributions of phlebotomine sand flies (Diptera: Psychodidae), vectors of leishmaniasis, in Iran. Acta Trop. 2014;132:131-9.
Postigo JA. Leishmaniasis in the World Health Organization Eastern Mediterranean Region. Int J Antimicrob Agents. 2010;36 Suppl 1:S62-5.
Khademvatan S, Salmanzadeh S, Foroutan-Rad M, Bigdeli S, Hedayati-Rad F, Saki J, et al. Spatial distribution and epidemiological features of cutaneous leishmaniasis in southwest of Iran. Alexandria J Med. 2016. doi: 10.1016/j.ajme.2016.03.001.
Singh N, Kumar M, Singh RK. Leishmaniasis: current status of available drugs and new potential drug targets. Asian Pac J Trop Med. 2012;5(6):485-97.
Tiuman TS, Santos AO, Ueda-Nakamura T, Filho BP, Nakamura CV. Recent advances in leishmaniasis treatment. Int J Infect Dis. 2011;15(8):e525-32.
Croft SL, Seifert K, Yardley V. Current scenario of drug development for leishmaniasis. Indian J Med Res. 2006;123(3):399-410.
Sundar S, More DK, Singh MK, Singh VP, Sharma S, Makharia A, et al. Failure of pentavalent antimony in visceral leishmaniasis in India: report from the center of the Indian epidemic. Clin Infect Dis. 2000;31(4):1104-7.
Albakhit S, Khademvatan S, Doudi M, Foroutan-Rad M. Antileishmanial activity of date (Phoenix dactylifera L) fruit and pit extracts in vitro. J Evid Based Complementary Altern Med. 2016. doi: 10.1177/2156587216651031.
Foroutan-Rad M, Hazrati Tappeh K, Khademvatan S. Antileishmanial and immunomodulatory activity of Allium sativum (Garlic): a review. J Evid Based Complementary Altern Med. 2015. doi: 10.1177/2156587215623126.
Khademvatan S, Eskandari A, Saki J, Foroutan-Rad M. Cytotoxic activity of Holothuria leucospilota extract against Leishmania infantum in vitro. Adv Pharmacol Sci. 2016;2016:8195381.
WHO. A report of the consultation meeting on traditional and modern medicine: harmonizing two approaches, 22–26 November 1999. Beijing, China, West Pacific Region. 2000.
Saki J, Khademvatan S, Pazyar N, Eskandari A, Tamoradi A, Nazari P. In vitro activity of Cordia myxa Mucilage extract against Leishmania major and L. infantum promastigotes. Jundishapur J Microbiol. 2015;8(3):e19640.
Yousefi E, Eskandari A, Gharavi MJ, Khademvatan S. In vitro activity and cytotoxicity of Crocus sativus extract against Leihmania major (MRHO/IR/75/ER). Infect Disord Drug Targets. 2014;14(1):56-60.
Sadeghi-Nejad B, Saki J. Effect of aqueous Allium cepa and Ixora brachiata root extract on Leishmania major promastigotes. Jundishapur J Nat Pharm Prod. 2014;9(2):e15442.
Allahdin S, Khademvatan S, Hashemitabar M, Eskandari A. In vitro activity of Camellia sinensis extracts against L. major and L. infantum promastigotes using the colorometric MTT assay. Urmia Med J. 2014;25(10):893-900.
Sadeghi-Nejad B, Saki J, Khademvatan S, Nanaei S. In vitro antileishmanial activity of the medicinal plant - Satureja khuzestanica Jamzad. J Med Plant Res. 2011;5(24):5912-5.
Harsha NS, Kumar G, Al-wesali MS, Rasool ST, Ibrahim FAS. Exploring the anti-leishmanial activity of Phoenix dactylifera (Date palm). Pharmacologyonline. 2009;3:791-9.
Althunibat OY, Hashim RB, Taher M, Daud JM, Ikeda M-A, Zali B. In vitro antioxidant and antiproliferative activities of three Malaysian sea cucumber species. Eur J Sci Res. 2009;37(3):376-87.
Adibpour N, Nasr F, Nematpour F, Shakouri A, Ameri A. Antibacterial and antifungal activity of Holothuria leucospilota isolated from Persian Gulf and Oman Sea. Jundishapur J Microbiol. 2014;7(1):e8708.
Bordbar S, Anwar F, Saari N. High-value components and bioactives from sea cucumbers for functional foods--a review. Mar Drugs. 2011;9(10):1761-805.
Kerr AM, Kim J. Phylogeny of Holothuroidea (Echinodermata) inferred from morphology. Zool J Linn Soc. 2001;133(1):63-81.
Dube A, Singh N, Saxena A, Lakshmi V. Antileishmanial potential of a marine sponge, Haliclona exigua (Kirkpatrick) against experimental visceral leishmaniasis. Parasitol Res. 2007;101(2):317-24.
Ben Kahla-Nakbi A, Haouas N, El Ouaer A, Guerbej H, Ben Mustapha K, Babba H. Screening of antileishmanial activity from marine sponge extracts collected off the Tunisian coast. Parasitol Res. 2010;106(6):1281-6.
Jimenez-Ruiz A, Alzate JF, Macleod ET, Luder CG, Fasel N, Hurd H. Apoptotic markers in protozoan parasites. Parasit Vectors. 2010;3:104.
Khademvatan S, Saki J, Gharavi MJ, Rahim F. Allium sativum extract induces apoptosis in Leishmania major (MRHO/IR/75/ER) promastigotes. J Med Plant Res. 2011;5(16):3725-32.
Khademvatan S, Gharavi MJ, Rahim F, Saki J. Miltefosine-induced apoptotic cell death on Leishmania major and L. tropica strains. Korean J Parasitol. 2011;49(1):17-23.
Feily A, Saki J, Maraghi S, Moosavi Z, Khademvatan S, Siahpoosh A. In vitro activity of green tea extract against Leishmania major promastigotes. Int J Clin Pharmacol Ther. 2012;50(3):233-6.
Khademvatan S, Adibpour N, Eskandari A, Rezaee S, Hashemitabar M, Rahim F. In silico and in vitro comparative activity of novel experimental derivatives against Leishmania major and Leishmania infantum promastigotes. Exp Parasitol. 2013;135(2):208-16.
Ghaffarifar F, Esavand Heydari F, Dalimi A, Hassan ZM, Delavari M, Mikaeiloo H. Evaluation of apoptotic and antileishmanial activities of Artemisinin on promastigotes and BALB/C mice infected with Leishmania major. Iran J Parasitol. 2015;10(2):258-67.
Huerta S, Goulet EJ, Huerta-Yepez S, Livingston EH. Screening and detection of apoptosis. J Surg Res. 2007;139(1):143-56.
Fumarola L, Spinelli R, Brandonisio O. In vitro assays for evaluation of drug activity against Leishmania spp. Res Microbiol. 2004;155(4):224-30.
Sugawara T, Zaima N, Yamamoto A, Sakai S, Noguchi R, Hirata T. Isolation of sphingoid bases of sea cucumber cerebrosides and their cytotoxicity against human colon cancer cells. Biosci Biotechnol Biochem. 2006;70(12):2906-12.
Janakiram NB, Mohammed A, Zhang Y, Choi CI, Woodward C, Collin P, et al. Chemopreventive effects of Frondanol A5, a Cucumaria frondosa extract, against rat colon carcinogenesis and inhibition of human colon cancer cell growth. Cancer Prev Res (Phila). 2010;3(1):82-91.
Fredalina BD, Ridzwan BH, Abidin AA, Kaswandi MA, Zaiton H, Zali I, et al. Fatty acid compositions in local sea cucumber, Stichopus chloronotus, for wound healing. Gen Pharmacol. 1999;33(4):337-40.
Yaacob H, Kim K, Shahimi M, Jamalulail S. Water extract of Stichopus sp. improves wound healing. J Perubatan Univ Kebangsaan Malays. 1994;16:19-29.
San Miguel-Ruiz JE, Garcia-Arraras JE. Common cellular events occur during wound healing and organ regeneration in the sea cucumber Holothuria glaberrima. BMC Dev Biol. 2007;7(1):115.
Zhao Q, Wang J-f, Xue Y, Wang Y, Gao S, Lei M, et al. Comparative study on the bioactive components and immune function of three species of sea cucumber. J Fishery Sci China. 2008;1:022.
4Ogushi M, Yoshie-Stark Y, Suzuki T. Apoptosis-inducing activity of hot water extracts from the sea cucumber in human colon tumor cells. Food Sci Technol Res. 2006;12(4):290-4.
Kalinin VI, Aminin DL, Avilov SA, Silchenko AS, Stonik VA. Triterpene glycosides from sea cucucmbers (holothurioidea, echinodermata). Biological activities and functions. Stud Nat Prod Chem. 2008;35:135-96.
Zou Z-R, Yi Y-H, Wu H-M, Wu J-H, Liaw C-C, Lee K-H. Intercedensides AC, three new cytotoxic triterpene glycosides from the sea cucumber Mensamaria intercedens Lampert. J Nat Prod. 2003;66(8):1055-60.
Singh N, Kumar R, Gupta S, Dube A, Lakshmi V. Antileishmanial activity in vitro and in vivo of constituents of sea cucumber Actinopyga lecanora. Parasitol Res. 2008;103(2):351-4.