Gharavi Mj, Nobakht M, Khademvatan Sh, Bandani E, Bakhshayesh M, Roozbehani M
Department of Medical Parasitology, Tehran University of Medical Sciences, Tehran, Iran & Research Institute for Islamic & Complementary Medicine, Tehran University of Medical Sciences, Tehran, Iran.
Iran J Parasitol. 2011 Aug;6(3):74-81.
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.
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.
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.
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.
本研究旨在揭示大蒜水提取物（AGE）对硕大利什曼原虫中IFNγ和iNOS基因表达的分子机制影响。
将硕大利什曼原虫前鞭毛体（MRHO/IR/75/ER）加入体外培养的J774细胞系中，细胞孵育72小时。向感染细胞中加入不同浓度的大蒜提取物（9.25、18.5、37、74、148毫克/毫升）。采用MTT法检测细胞增殖情况。孵育72小时后，收集上清液，并从感染细胞中提取总RNA。通过RT-PCR法研究IFNγ和iNOS基因的表达。
孵育3天后的比色MTT分析显示大蒜提取物具有细胞毒性作用，IC50为37毫克/毫升。此外，RT-PCR检测IFNγ和iNOS基因表达表明，大蒜提取物可导致这些基因在感染硕大利什曼原虫的J774细胞系中过表达。
大蒜提取物对感染的J774细胞系具有细胞毒性作用。此外，大蒜可通过提高IFNγ和iNOS基因的表达来改善细胞免疫这一假设得到了证实。
