Evaluation of synergistic therapeutic effect of shark cartilage extract with artemisinin and glucantime on visceral leishmaniasis in BALB/c mice

Document Type : Original Article

Authors

1 Department of Parasitology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Deputy of Research, Ardabil University of Medical Sciences, Ardabil, Iran

3 Department of Immunology, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran

4 Department of Virology, Iranian Blood Transfusion, Tehran, Iran

Abstract

Objective(s): Because leishmaniasis is related to the impaired functioning of T-cells, the use of an immunomodulator can increase the efficacy of antileishmanial therapy in visceral leishmaniasis. In this study, we used shark cartilage extract with artemisinin and glucantime against visceral leishmaniasis in BALB/c mice, and evaluated the synergistic therapeutic effect.
Materials and Methods: The culturing method and quantitative real-time PCR by using the kDNA gene was used to detect parasite loads in the spleen and liver. INF-γ and IL-4 cytokine levels and survival rates were assayed.
Results: The drug therapy with target drugs reduced parasite burden in the spleen and liver significantly. Although parasite burden was lower in the artemisinin treated group than in the glucantime treated group (P<0.05). The mice survival rate records, throughout the experimental period, showed highly significant survival rates in the test groups compared to the control group (P<0.001). The results of cytokine assay in mice treated with glucantime-shark cartilage extract combination indicated significant increases of IFNγ and IL-4 (P<0.05). Although the increase of IFNγ was more notable than IL-4. The synergistic therapeutic effect is shown in all groups except in the group treated with shark cartilage extract-artemisinin combination. The IFN-γ in glucantime-shark cartilage extract combination treated group was higher than in other groups (P<0.05). The survival rate in this group was more than in other groups too (P<0.05).
Conclusion: Combination therapy with shark cartilage extract as an immunomodulator can increase antileishmanial effects of antimony drugs in VL treatment.

Keywords

Main Subjects


1. WHO. Accelerating work to overcome the global impact of neglected tropical diseases: a roadmap for implementation. 2012, Geneva, Switzerland. WHO/HTM/NTD/2012. World Health Organization, Geneva.
2. Alvar J, Vélez ID, Bern C, Herrero M, Desjeux P, Cano J, et al. WHO leishmaniasis control team: Leishmaniasis worldwide and global estimates of its incidence. PLoS One 2012; 7:e35671.
3. Dujardin JC, Campino L, Cañavate C, Dedet JP, Gradoni L, Soteriadou K, et al. Spread of vector-borne diseases and neglect of  leishmaniasis. Europe Emerg Infect Dis 2008; 14:1013–1018.
4. Frezard F, Demicheli C, Ribeiro RR. Pentavalent antimonials: New perspectives for old drugs. Molecules 2009; 14:2317-2336.
5. Meshnick SR. Artemisinin: mechanisms of action, resistance and toxicity. Int J Parasitol 2002; 32:1655-1660.
6. Sundar S, Chatterjee M. Visceral leishmaniasis-current therapeutic modalities. Indian J Med Res 2006; 123:345-352.
7. Yardley V, Craft SL. A comparison of the activities of three amphotericin B lipid formulations against experimental visceral and cutaneous leishmaniasis. Int J Antimicrob Agents 2000; 13:243-248.
8. Sundar S, Murray HW. Availability of miltefosine for the treatment of Kala-azar in India. Bull World Health Organ 2005; 83:394-395.
9. Yang DM, Liew FY. Effects of qinghaosu (artemisinin) and its derivatives on experimental cutaneous leishmaniasis. Parasitology 1993; 106:7–11.
10. Sen R, Chatterjee M. Plant-derived therapeutics for the treatment of leishmaniasis. Phytomedicine 2011; 18:1056–1069.
11. Meshnick SR. From quinine to qinghaosu: Historical perspectives in malaria: parasite biology, pathogenesis, protection. 1st ed. Washington DC: ASM Press; 1998. P. 341–353.
12. Olsson ME, Olofsson LM, Lindahl AL, Lundgren A, Brodelius M, Brodelius PE. Localization of enzymes of artemisinin in biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry 2009; 70:1123–1128.
13. Avery MA, Muraleedharan KM, Desai PV, Bandyopadhyaya AK, Furtado MM, Tekwani BL. Structure-activity relationships of the antimalarial agent artemisinin. 8. design, synthesis, and CoMFA studies toward the development of artemisinin-based drugs against leishmaniasis and malaria. J Med Chem 2003; 46:4244-4258.
14. Ganguly S, Bandyopadhyay S, Bera A, Chatterjee M. Anti-promastigote activity of an ethanolic extract of leaves of Artemisia indicia. Indian J Pharmacol 2006; 38:64–65.
15. Ganguly S, Bandyopadhyay S, Sarkar A, Chatterjee M. Development of a semi-automated colorimetric assay for screening anti-leishmanial agents. J Microbiol Methods 2006; 66:79–86.
16. Isavand– Heidari F, Ghaffarifar F, Dalimi A, Mortazavi- Dehkordi N, Ghasemi- Nikoo S. In vitro study of the effect of artimisinin on promastigotes and amastigotes of Leishmania major. Pathobiology (TMUJ) 2012; 15:33-43. (Persian)
17. Sen R, Bandyopadhyay S, Dutta A, Mandal G, Ganguly S, Saha P, et al. Artemisinin triggers induction of cell-cycle arrest and apoptosis in Leishmania donovani promastigotes. J MED Microbiol 2007; 56:1213–1218.
18. Ghaffarifar F, Esavand-Heydari F, Dalimi A, Hassan ZM, Delavari M, Mikaeiloo M. Evaluation of apoptosis and anti-leishmanial activities of artemisinin on promastigotes and BALB/c mice infected with Leishmania major. Iran J Parasitol 2015; 10:258-267.
19. Sen R, Ganguly S, Saha P, Chatterjee M. Efficacy of artemisinin in experimental visceral leishmaniasis. Int J Antimicrobial Agents 2010; 36:43–49.
20. Kaye PM, Svensson M, Ato M, Maroof A, Polley R, Stager S, et al. The immunopathology of experimental visceral leishmaniasis. Immunol Rev 2004; 201:239–253.
21. Sharma U, Singh S. Immunobiology of leishmaniasis. Indian J Exp Biol 2009; 47:412–423.
22. Anonymous. Shark cartilage. Drug Store News for the Pharmacist 1995;5:41.
23. Dvorkin L, Thu L, Gerzenshtein S. Shark Cartilage. Longwood Herbal Task Force. Http://www.mcp.edu/herbal/default.htm, Revised September 14, 1999.
24. Brem H, Folkman J. Inhibition of tumor angiogenesis mediated by cartilage. J Exp Med 1975; 141:427-439.
25. Kestin M, Miller L, Littlejohn G, Wahlqvist M. The use of unproven remedies for rheumatoid arthritis in Australia. J MED Aust 1985; 143:516-518.
26. Dupont E, Savard PE, Jourdain C, Juneau C, Thibodeau A, Ross N, et al. Antiangiogenic properties of a novel shark cartilage extract: potential role in the treatment of psoriasis. J Cutan Med Surg 1998; 2:146-152.
27. Merly L, Simjee S, Smith SL. Induction of inflammatory cytokines by cartilage extracts. Int Immunopharmacol 2007; 7:383-391.
28. Feyzi R, Hassan ZM, Mostafaie A. Modulation of CD4(+) and CD8(+) tumor infiltrating lymphocytes by a fraction isolated from shark cartilage: shark cartilage modulates anti-tumor immunity. Int Immunopharmacol 2003; 3:922-926.
29. Molaie S, Ghaffarifar F, Hasan ZM, Dalimi A. Enhancement effect of artemisinin with glucantime and shark cartilage extract on the killing factors and apoptosis of Leishmania infantum in vitro condition. IJPR 2017. ID: 10131 (Accepted, not published)
30. Nemati S, Nahrevanian H, Haniloo A, Farahmand M. Investigation on nitric oxide and C- reactive protein involvement in anti-leishmanial effects of artemisinin and glucantime on cutaneous leishmaniasis. Adv Stud Biol 2013; 5:27–36.
31. Hassan ZM, Feyzi R, Sheikhian A, Bargahi A, Mostafaie A, Mansouri K, et al. Low molecular weight fraction of shark cartilage can modulate immune responses and abolish angiogenesis. Int Immunopharm. 2005; 5:961– 970.
32. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem1976; 72:248–254.
33. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685.
34. Mortazavi- Dehkordi N, Ghaffarifar F, Hasan ZH, Esavand- Heydari F. In vitro and in vivo studies of anti-leishmanial effect of artemether on Leishmania infantum. Jundishapur J Microbiol 2013; 6:e6379.
35. Ulbricht C, Boon H, Philippe-Szapary O, Basch E. Shark cartilage monograph: A clinical decision support tool. J Herbal Pharmacotherap 2002;  2:7-13.
36. Zheng L, Ling P, Wang Z, Niu R, Hu CH, Zhang T, et al. A novel polypeptide from shark cartilage with potent anti-angiogenic activity. Cancer Biol Ther 2007;  6:775-780.
37. Ahmed S, Colmenares M, Soong L, Goldsmith-Pestana K, Munstermann L, Molina R, et al. Intradermal infection model for pathogenesis and vaccine studies of murine visceral leishmaniasis. Infect Immun 2003;71:401–410.
38. Srivastava A, Sweat JM, Azizian A, Vesely B, Kyle DE. Real-time PCR to quantify Leishmania donovani in hamsters. J Parasitol 2013; 99:145-50.
39. Verma S, Kumar R, Katara GK, Singh LC, Negi NS, Ramesh V, et al. Quantification of parasite load in clinical samples of leishmaniasis patients: IL-10 level correlates with parasite load in visceral leishmaniasis. PLoS One 2010; 5: e10107.
40. Maroufi Y, Ghaffarifar F, Dalimi A, Sharifi Z. Leishmaniasis treated with Cantharidin. Jundishapur J Microbiol 2014; 7: e10907.
41. Johan-van G, Manica B, Filip M, Jorge A, Lutgarde L, Marleen B. Combination therapy for visceral leishmaniasis. Lancet Infect Dis 2010; 10:184–194. 42. Gazanion E, Vergnes B, Seveno M, Garcia D, Oury B, Ait-Oudhia K, et al. In vitro activity of nicotinamide anti-leishmanial drug combinations. Parasitol Int 2011; 60:19-24.
43. Alvar J, Croft S, Olliaro P. Chemotherapy in the treatment and control of leishmaniasis. Adv Parasitol 2006; 61: 223–274.
44. Golenser J, Waknine JH, Krugliak M, Hunt NH, Grau GE. Current perspectives on the mechanism of action of artemisinins. Int J Parasitol 2006; 36:1427-1441.
45. Wang J, Zhou H, Zheng J, Cheng J, Liu W, Ding G, et al. The antimalarial artemisinin synergizes with antibiotics to protect against lethal live Escherichia coli challenge by decreasing proinflammatory cytokine release. Antimicrob Agents Chemother  2006; 50:2420–2427.
46. Vanessa Ribeiro M, Luís Cláudio LJ, Rossy-Eric PS, Luis Douglas MS, Bruno Araújo SP, Maria Norma M, Antonio Marcus AP, Silma Regina FP. Meglumine antimoniate (Glucantime) causes oxidative stress-derived DNA damage in BALB/c mice infected by Leishmania infantum. Antimicrob Agents Chemother 2017; 81:1-9.
47. Piu S, Debanjan M, Mitali Ch. Immunomodulation by chemotherapeutic agents against leishmaniasis. Int Immunopharmacol 2011;11: 1668–1679.
48. Handman E, Bullen DV. Interaction of leishmania with the host macrophage. Trends Parasitol 2002;18:332-334.