Articles

The Effect of Garlic Extract on Expression of INFγ And Inos Genes in Macrophages Infected with Leishmania major

Abstract

Background: The study was aimed to show the effect of molecular mechanism of Aqueous Garlic Extract (AGE) on expression of IFNγ and iNOS genes in Leishmania major.

Methods: Leishmania major promastigotes (MRHO/IR/75/ER) were added to the in-vitro cultured J774 cell line, the cells were incubated for 72 hours. Various concentrations of garlic extract (9.25, 18.5, 37, 74, 148 mg/ml) were added to the infected cells. MTT assay was applied for cellular proliferation. After 72 hours of incubation, supernatants were collected and total RNA was extracted from the infected cells. The express of IFNγ and iNOS genes were studied by RT-PCR method.

Results: The colorimetric MTT assay after 3 days of incubation showed cytotoxic effect of garlic extract with an IC50 of 37 mg/ml. In addition, IFNγ and iNOS genes expression by RT-PCR indicated that garlic extract lead to over expression of these genes in J774  cell line infected with L. major.

Conclusion: Garlic extract exerts cytotoxic effect on infected J774 cell line. In addition, the hypothesis that garlic can improve cellular immunity with raising the expression of IFNγ and of iNOS genes con­firmed.

Modabber F. Leishmaniasis. In:. Walgate R, Simpson K, editors. Tropical diseases research progress Geneva: World Health Organization. 1995. p.77–88.

Khademvatan SH, Gharavi MJ, Rahim F, Saki J. Miltefosine induced apoptotic cell death on Leishmania major and L. tropica strains. Korean J Parasitology. 2011; 49(1):1-8.

Hepburn NC, Siddique I, Howie AF, Beckett GJ, Hayes PC. Hepatotoxicity of sodium stibogluconate therapy for American cutaneous leishmaniasis. Trans R Soc Trop Med Hyg. 1994;88:453–455.

Delaha EC, Garagusi VF. Inhibition of mycobacteria by garlic extract (Allium sativum). J Antimicrob Chemother. 1985; 27(4): 485-48.

Appleton J, Tansey MR. Inhibition of growth of zoopathogenic fungi garlic extract. J Mycologia. 1975; 67:882-5.

Kandil OM, Abdullah TH. Garlic and the immune system in humans: Its effect on killer cells. Fed Proc. 1987; 46: 41-6.

Panosian CB, Sypek JP, Wyler DJ. Cell contact-mediated macrophage activation for antileishmania defence. I. Lymphocyte effector mechanism that is contact dependent and noncytotoxic. J Immunol. 1984; 133:3358-65.

Ghazanfari T, Hassan ZM, 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): 491-5.

Khademvatan SH, Gharavi MJ, Akhlaghi L. Induction of apoptosis by miltefosine in Iranian strain of Leishmania infantum promastigotes. Iran J Parasitol. 2009; 4(2):23-30.

Olliaro PL, Bryceson AD. Practical progress and new drugs for changing patterns of leishmaniasis. Parasitol Today. 1993;9:323–32.

Herwaldt BL, Berman JD. Recommendations for treating leishmaniasis with sodium stibogluconate (Pentostam) and review of pertinent clinical studies. AmJ TropMedHyg 1992;46:296–306.

Ahmadi-Renani K, Mahmoodzadeh A, Cheraghali AM, Esfahani A. Effect of Garlic Extract on Cutaneous Leishmaniasis and the Role of Nitric Oxide. IJMS. 2002; 27(3): 97-100.

Khalid NM, Mohomed HE, Toum AM, Mubark MA, Magzoub MA. Treatment of Cutaneous leishmaniasis with some Local Sudanese Plants (Neem, Garad & Garlic). Türkiye Parazitoloji Dergisi 2004; 28 (3): 129-132.

Mattner J, Wandersee-Steinhäuser A, Pahl A, Röllinghoff M, Majeau GR, Hochman PS, Bogdan C. Protection against Progressive Leishmaniasis by IFN-β1. J Immunol. 2004;172: 7574-82.

Bogdan C, Gessner A, Rollinghoff M. Cytokines in leishmaniasis: a complex network of stimulatory and inhibitory interactions. J Immunobiology. 1993; 189:356-361.

Stenger S, Donhaauser N, Thuring H, Rollinghoff M, Bogdan C. Reactivation of latent leishmaniasis by inhibition of inducible nitric oxide synthase. J Exp Med. 1996; 183:1501-6.

Ruhland A, Kima PE. Activation of PI3K/Akt signaling has a dominant negative effect on IL-12 production by macrophages infected with Leishmania amazonensis promastigotes. Exp Parasitol. 2009; 122 (1):28–36.

Gazzinelli RT, Oswald IP, James SL, Sher A. IL-10 inhibits parasite killing and nitrogen oxide production by IFN-gamma-activated macrophages. J Immunol. 1992; 148:1792–6.

Umar S, van der Laarse A. Nitric oxide and nitric oxide synthase isoforms in the normal, hypertrophic and failing heart. J Mol Cell Biochem. 2010; 333(2):191-201.

Dutta A, Mandal G, Mandal C, Chatterjee M. In vitro antileishmanial activity of Aloe vera leaf exudate: a potential herbal therapy in leishmaniasis. Glycoconj J. 2007; 24(1):81-6.

Trun W, Kiderlen AF, Kolodziej H. Nitric oxide synthase and cytokines gene expression analyses in Leishmania- infected RAW 264.7 cells treated with an extract of Pelargonium sidoides. Phytomedicine. 2006; 13 (8):570-5.

Isaac-Marquez AP, Lezama-Davila CM. Detection of pathogenic bacteria in skin lesions of patients with chiclero’s ulcer. Reluctant response to antimonial treatment. Mem Inst Oswaldo Cruz. 2003;98:1093–5.

Tsao SM, Hsu C, Yin MC. Garlic extract and two diallyl sulphides inhibit methicillin resistant Staphylococ-cus aureus infection in BALB/cA mice. J Antimicrob Chemother. 2003; 52: 974–980.

Files
IssueVol 6 No 3 (2011) QRcode
SectionArticles
Keywords
Aqueous Garlic Extract (AGE) Leishmania major Macrophage Gene Expression

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Gharavi M, Nobakht M, Khademvatan S, Bandani E, Bakhshayesh M, Roozbehani M. The Effect of Garlic Extract on Expression of INFγ And Inos Genes in Macrophages Infected with Leishmania major. Iran J Parasitol. 1;6(3):74-81.