Butyric acid, the product of bacterial fermentation of complex carbohydrates in the large intestine, is a preferred energy source for normal colonocytes and stimulator of their proliferation in vivo. In contrast, butyrate has been shown to inhibit proliferation and induce apoptosis in a number of colorectal tumor cell lines. Among the possible mechanisms by which butyrate may exert anti-carcinogenic effects, its antioxidant activity has recently been suggested. Reactive oxygen species (ROS) are involved in many physiological and pathological processes, such as cell proliferation control, apoptosis and gene expression induction. An overproduction of ROS or their limited inactivation may cause cellular damage resulting in genomic changes and carcinogenesis process induction.

The aim of this study was to evaluate the impact of sodium butyrate (NaB) on antioxidative enzymes glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity in colon cancer cell line Caco-2. Cells (1x10⁶/dish) were cultured in Minimum Essential Medium supplemented with 10% fetal bovine serum, 100 units/ml penicillin, 100 μg/ml streptomycin, and 10 mM HEPES at 37 ^oC in a humidified atmosphere with 5% CO₂. The cells were incubated for 3 days to adhere to the plates, and then they were exposed for 4 days to 1mM and 10 mM NaB. Afterwards, the adherent cells were washed with phosphate-buffered saline, harvested and sonicated. Cell supernatants were used for GPx and SOD activity assay with the use of Ransod and Ransel kits (RANDOX). Morphological analysis of cell nuclei was evaluated based on their characteristic changes visualized after staining with 5 μg/ml acridine orange and observed under fluorescence microscope (Olympus BX-60). Cellular differentiation of NaB-treated cells was evaluated by measuring alkaline phosphatase (AP) activity.

The results of this study showed that butyrate changed GPx activity in Caco-2 cells. The highest activity of GPx was observed in colonocytes incubated with 1 mM butyrate.

The highest SOD and AP activities were observed in Caco-2 cells treated with 10 mM butyrate.

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