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Published on: March 16, 2018
The Effect of Garlic Extract on Expression of INFγ And Inos Genes in Macrophages Infected with Leishmania major
Mj Gharavi1, M Nobakht, Sh Khademvatan
1Department of Medical Parasitology, Tehran University of Medical Sciences, Tehran, Iran & Research Institute for Islamic & Complementary Medicine, Tehran University of Medical Sciences, Tehran, Iran.
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.
Insights
Aqueous Garlic Extract (AGE) demonstrated a cytotoxic effect on Leishmania major-infected cells and boosted the expression of IFNγ and iNOS genes, suggesting enhanced cellular immunity.
Area of Science:
- Immunology
- Molecular Biology
- Parasitology
Background:
- Investigating the molecular mechanisms of Aqueous Garlic Extract (AGE) against Leishmania major infections.
- Understanding the impact of AGE on host immune gene expression.
Purpose of the Study:
- To determine the effect of AGE on the expression of IFNγ and iNOS genes in Leishmania major-infected cells.
- To evaluate the cytotoxic effects of AGE on infected host cells.
Main Methods:
- In vitro culture of Leishmania major promastigotes with J774 cell line.
- Treatment with various concentrations of AGE.
- MTT assay for cellular proliferation and cytotoxicity.
- RT-PCR for gene expression analysis of IFNγ and iNOS.
Main Results:
- AGE exhibited a cytotoxic effect with an IC50 of 37 mg/ml.
- RT-PCR analysis revealed overexpression of IFNγ and iNOS genes in infected cells treated with AGE.
- AGE treatment led to increased expression of key immune response genes.
Conclusions:
- Aqueous Garlic Extract (AGE) possesses cytotoxic properties against Leishmania major-infected J774 cells.
- AGE enhances cellular immunity by upregulating IFNγ and iNOS gene expression.
- Garlic extract shows potential as an immunomodulatory agent in leishmaniasis treatment.
