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Evaluating the effect of diallyl sulfide on regulation of inflammatory mRNA expression in 3T3L1 adipocytes and RAW 264.7 macrophages during ethanol treatment

Abstract Diallyl sulfide (DAS) has been studied extensively for its alleged role as an anticancer and protective agent. Alcohol influences and effects on human health have been extensively studied. However, investigations toward developing and testing therapeutic agents that can reduce the tissue injury caused by ethanol are scarce. In this backdrop, this study was designed to explore the potential effect of DAS in reducing alcohol induced damage of 3T3L1 adipocytes and RAW 264.7 macrophages. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed to determine the DAS effect on cell viability. Reactive oxygen species (ROS) production was assessed by flow cytometer. Expression of inflammatory genes was studied by the qRT-PCR method. Our study results showed that DAS at concentrations less than 200 μM was not toxic to the cells and the viability of ethanol-exposed 3T3L1 adipocyte cells was found to be significantly increased when ethanol-exposed cells were treated with DAS. Further, treatment of ethanol-exposed 3T3L1 cells with 100 μM DAS for 24 h was found to reduce ethanol induced ROS production, expression of pro-inflammatory cytokines, and enhance anti-inflammatory cytokine production in the cells. Also, 100 μM DAS was found to increase the expression of M2 phenotype-specific genes in ethanol-exposed RAW 264.7 macrophage cells. Further, 100 μM DAS also improved the levels of lipid accumulation in 3T3L1 adipocytes that was down-regulated by ethanol exposure. Taken together, our study results imply that DAS may be effective in reducing ethanol induced injury of cells thereby suggesting its potential to be used in drug formulations.