The viability of leukocytes and reactive oxygen species generation by them in rats with chronic colitis

Abstract

The aim: To assess reactive oxygen species production by leukocytes and their viability in rats with chronic colitis. Materials and methods: Reactive oxygen species production was estimated in leukocytes, isolated from rats with Dextran Sulfate Sodium-induced chronic colitis and control rats, by flow cytometry using the fluorescent probe 2’,7’-dichlorodihydrofluorescein diacetate. Leukocyte viability and apoptosis stages were assessed by flow cytometry using annexin V and 7-aminoactinomycin D staining. White blood cell counting was carried out with using Hematology Analyzer. Results: The increased fluorescence intensity of 2’,7’-dichlorofluorescein in viable leukocytes by 36.7% was revealed in rats with chronic colitis compared control rats. A significant decrease in the percentage of viable cells and an increase in apoptotic cells were found compared to intact animals. Leukocytes, granulocytes, monocytes, lymphocytes counts in blood of experimental group animals were significantly higher compared to control those. Conclusions: Our findings indicate that Dextran Sulfate Sodium-induced chronic colitis increases an intracellular production of reactive oxygen species by leukocytes. Despite of increased leukopoiesis it reduces viability of white blood cells and promotes their apoptosis via stimulation of oxidative stress.