t is known that alcohol influences on human health and its effects are widely studied. Adipose tissue (AT) may be an important target of ethanol action. Data about the effect of ethanol on cytokine and pro-inflammatory enzymes` gene expression in adipocytes are scarce. The aim of the present study was to establish its effect on transcription of inflammatory markers IL-6, TNF-α, COX-2 and iNOS in cultured 3T3-L1 cells. MTT test was performed in order to determine ethanol effect on cell viability. Expression of selected genes was measured using two step quantitative PCR. Results were analyzed using 2-ΔΔCt method. Exposure of 3T3-L1 cells to increasing ethanol concentrations (0.125%-0.625%) resulted in gradual cell viability decline. Expression of all the genes studied was up-regulated upon ethanol treatment. 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-α, iNOS and COX-2, respectively, as compared with untreated controls. mRNA levels of the respective genes highly correlated with the applied ethanol concentration. In conclusion, our results show that ethanol treatment may increase the inflammatory potential of AT up-regulating expression of inflammatory cytokines IL-6 and TNF-α and inflammatory enzymes iNOS and COX-2 on transcription level.
Scripta Scientifica Medica 2012; 44(2): 31-35.
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