اثر عصاره سیر بر بیان ژن‌های iNOSو IFNγ در رده سلولی J774 آلوده به لیشمانیا ماژور

  Background and Objectives : The protozoa of leishmania lead to cutaneus and visceral leishmaniasis. Activation macrophages phagocyte parasites and this mechanism can be done with production of active radicals oxygen and nitric oxide, That is done through immunomodulator , as a basic process for production of the new drug. In this study, the effect of garlic extract as an immunomodulator on J774 cells infected with Leishmania major and iNOS gene expression and IFNγ has been investigated.   Materials and Methods: Leishmania major promastigotes (MRHO/IR/75/ER) were added to the in-vitro cultured macrophages (J774 cells) and these 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 prolifration. After 72 hours of incubation, supernatants were collected and total RNA was extracted from the infected cells. The expression of IFNγ and INOS genes were studied by RT-PCR method. Untreated sample with garlic was used as control. All the experiments were repeated at least three times, and representative results were analyzed. Statistical significance (P < 0.05) was analyzed by Student’s t-test using SPSS version16.   Results: The result of this study, using MTT method showed that, the IC50 concentration of garlic extract was 37 mg/ml. Also the expression of IFNγ and iNOS genes by RT-PCR indicated that garlic extract lead to rise of expression of these genes in the J774 cells infected with L.major.   Conclusion: Based on the findings and importance of the cellular immunity cytokines in developing and importance of the responses, the hypothesis that the effect of AGE in cellular immunity is due to rising the IFNγ cytokines and expression of iNOS genes is confirmed.   Key words: Aqueous Garlic Extract (AGE), IFNγ and iNOS genes, Leishmania major, J774 cell line   Funding: This study was supported by Research Institute for Islamic & Complementry Medicine of Tehran University of Medical Sciences.   Conflict of interest: None declared.   Ethical approval: The Ethics Committee of Research Institute for Islamic & Complementry Medicine of Tehran University of Medical Sciences approved the study.   How to cite this article : Gharavi MJ, Nobakht M, Khademvatan Sh, Bandani E, Roozbehani M, Bakhshayesh M, Samadi Koochaksaraei A . he Effect of Garlic Extract on Expression of IFN-γ and iNOS Genes in Macrophages Infected with Leishmania Major. J Rafsanjan Univ Med scie 2012 11(4): 313-22. [Farsi] .