(MDA), a marker of oxidative stress, was measured by HPLC and the thiobarbituric acid method.Įxpression of inflammatory genes and lipogenic genes was studied by qRT-PCR and Western blotting.
Of human primary adipocytes on DAS treatment was assessed by MTT assay.
We also studied DAS effect on isolated human primary adipocytes. During the last week of feeding protocol, the EtOH-fed rat group was given 200 mg/kg body Control rats were pair-fed with isocaloric diet containing maltodextrin instead ofĮtOH. Methods: Male Wistar rats were fed with 36% Lieber–DeCarli liquid diet containing ethanol This study was designed to test theĮfficacy of diallyl sulfide (DAS) in regulating the alcohol-induced outcomes on adipose tissue. Known to play a significant role in alcohol-induced tissue damage. Adipose tissue apart from functioningĪs an endocrine organ secretes several hormones and cytokines known as adipokines that are Several cellular pathways contribute to alcohol-mediated tissue injury. 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.īackground: Alcohol consumption is the fourth leading cause of death and disability worldwide. Further, 100 μM DAS also improved the levels of lipid accumulation in 3T3L1 adipocytes that was down-regulated by ethanol exposure. Also, 100 μM DAS was found to increase the expression of M2 phenotype-specific genes in ethanol-exposed RAW 264.7 macrophage cells. 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. 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. Expression of inflammatory genes was studied by the qRT-PCR method. Reactive oxygen species (ROS) production was assessed by flow cytometer. MTT assay was performed to determine the DAS effect on cell viability. 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. However, investigations toward developing and testing therapeutic agents that can reduce the tissue injury caused by ethanol are scarce. Alcohol influences and effects on human health have been extensively studied. In conclusion, our results show that ethanol treat-ment may increase the inflammatory potential of AT up-regulating expression of inflammatory cytokines IL-6 and TNF-a and inflammatory enzymes iNOS and COX-2 on transcription level.ĭiallyl sulfide (DAS) has been studied extensively for its alleged role as an anticancer and protective agent. mRNA levels of the respective genes highly correlated with the applied ethanol concentration. In cells treated with 0,5% ethanol, there was a 4,8-(p<0,001), 6,3-(p<0,001), 5,5-(p<0,05) and 3,6-(p<0.05) fold increase of mRNA levels of IL-6, TNF-a, iNOS and COX-2, respectively, as compared with untreated controls. Expression of all the genes studied was up-regulated upon ethanol treatment. Exposure of 3T3-L1 cells to increasing ethanol concentrations (0,125%-0,625%) resulted in gradual cell viability decline. Results were analyzed using 2 -DDCt method. Expression of selected genes was mea-sured using two step quantitative PCR. MTT test was performed in order to determine ethanol effect on cell viability. The aim of the present study was to establish its effect on transcription of inflammatory markers IL-6, TNF-a, COX-2 and iNOS in cultured 3T3-L1 cells. Data about the effect of ethanol on cytokine and pro-inflammatory enzymes' gene expression in adipocytes are scarce. Adipose tissue (AT) may be an important target of ethanol action. It is known that alcohol influences on human health and its effects are widely studied.
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