The impact of humus water on inflammatory cytokines of endothelial cells cultured under conditions of hyperglycemia

Abstract

Introduction: Diabetes has become an epidemic and is a risk factor of cardiovascular disease. Hyperglycemia in diabetes is responsible for damaging of the endothelium and increases inflammation on the surface of the vascular lining. The inflammatory process in diabetes is associated with the secretion of inflammatory cytokines by endothelial cells, e.g., tumor necrosis factor-alpha (TNF-α) and interleukin 6 (IL-6) and by a reduction of cell proliferation.Humus waters (HW) containing humus acids with sources in Poland are originally bacteriologically pure, physically and chemically stable. Because of the physicochemical properties of humus acids of adsorption, complexation and ion exchange they present numerous therapeutic activities including anti-inflammatory. Data regarding its molecular mechanisms underlying these phenomena are scarce. The present study reveals that humus water exerts important beneficial effects on endothelial cells under conditions of hyperglycemia.Material and methods: Endothelial cells (HUVEC line) were derived from human umbilical veins using the enzymatic method with collagenase. Cells were cultured in M199 media in accordance the standard method. The experiment was repeated three times with three independent cells isolations. The study was conducted in four groups: 1-control group -culture medium without impact of glucose and humus water; 2 -an appropriate volume of humus water was added to obtain its 1% solution in the culture medium; 3-(30 mM/L) glucose added to the culture medium to imitate hyperglycemic condition; 4 -glucose, humus water in the medium. At the end of experiment, conditioned medium from each well of culture plates was collected and the concentration of TNF-α and IL-6 in the supernatant was measured by ELISA test (Diaclone) according to the manufacturer's instructions.