Secretion and gene expression of proatherosclerotic cytokines IL6 and IL8 by endothelial cells exposed to mutagen

Atherosclerosis is a leader in morbidity and mortality among all cardiovascular pathologies both in Russia and around the world. Nowadays there are evidences that somatic mutations in addition to classical risk factors can play a role in the development of endothelial dysfunction - the initial stage of atherogenesis, and atherosclerotic plaques are characterized by increased level of various DNA adducts. At the same time, a fundamental base of the contribution of mutagenesis to atherogenesis is still unstudied thought it is important for industrial regions with high genotoxic risk The aim of this research was to study the secretion and gene expression of proatherosclerotic cytokines IL-6 and IL-8 by primary human coronary- and internal thoracic artery endothelial cells characterized by different sensitivity to atherogenesis and exposed to alkylating mutagen mitomycin C. Concentration of the studied cytokines in culture medium and mRNA level of the corresponding genes were measured using quantitative polymerase chain reaction and enzyme-linked immunosorbent assay at two time points - immediately after 6 hours of cell incubation with mitomycin C and after 6 hours of cell incubation with mutagen followed by 24 hours incubation in the complete media without mitomycin C. It was found that at the first time point, a significant decrease in the secretion and gene expression of IL-8, as well as a decrease in the mRNA level of the IL-6 gene in the exposed culture was discovered. At the second time point, on the contrary, the concentration and mRNA level of IL-8, as well as expression of the IL6 gene sharply increased in cultures exposed to mitomycin C in comparison with control (p <0.01), and the coronary artery cells were more sensitive to mutagenic effects than the cells of the internal thoracic artery. Thus, for the first time in in vitro experiment, mutagen-induced changes in the secretion and gene expression of proatherosclerotic cytokines IL-6 and IL-8 in primary human endothelial cells derived from various arteries have been shown.

mutagenesis, atherosclerosis, endothelial cells, coronary artery, internal thoracic artery, mitomycin C, cytokines, gene expression, ELISA

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