Gutkha chewing induces oxidative disturbances and cellular DNA damage in humans

Abstract

Chewing of gutkha is a risk factor for several diseases. The purpose of this study was to determine the effects of gutkha on oxidative damage and antioxidant enzyme status in serum, lymphocyte, and neutrophils. Male subjects were randomly selected from university students (21–24 years) in a rural community in the District of Midnapore, West Bengal, India. In order to examine oxidative damage and antioxidant enzyme status, superoxide radical generation (), nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was measured in lymphocytes and neutrophils; and in serum, neutrophils, and lymphocytes the levels of lipid peroxidation (MDA), reduced glutathione (GSH), and oxidized glutathione (GSSG) as well as activities of lactate dehydrogenase (LDH), catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) were measured. Deoxyribonucleic acid (DNA) damage in lymphocytes and neutrophils was determined by diphenylamine (DPA) assay and 1.2% gel electrophoresis. There was a significant increase in generation, NADPH oxidase, LDH activity, and DPA in lymphocytes and neutrophils, and a rise in the levels of MDA and GSSG in serum, lymphocytes, and neutrophils of gutkha chewers. GSH levels, redox ratio, and the activity of SOD, CAT, GPx, and GR of serum, lymphocyte, and neutrophils in gutkha chewers were significantly decreased. Agarose gel electrophoresis showed a smeary pattern of DNA. These findings suggest that chewing of gutkha produced oxidative stress and impaired antioxidant defense system.