Protective effect of catechin on pentachlorophenol-induced cytotoxicity and genotoxicity in isolated human blood cells.

Abstract

Pentachlorophenol (PCP) is an organochlorine compound that is used as pesticide, biocide, and wood preservative. PCP is highly toxic and carcinogenic. It has been detected in food and several consumable products. The toxicity of PCP is thought to be due to generation of oxidative stress in cells. We examined whether the dietary antioxidant catechin can attenuate or protect human erythrocytes and lymphocytes against PCP-induced cytotoxicity and genotoxicity, respectively. Human erythrocytes were treated with increasing concentrations of catechin (0.05-2.5mM) for 30min followed by addition of 0.75mM PCP and further incubation for 4h at 37°C. Hemolysates were prepared and assayed for various biochemical parameters. Treatment with PCP alone increased the generation of reactive oxygen and nitrogen species, lipid and protein oxidation, and damaged the plasma membrane, when compared to PCP untreated (control) cells. It significantly decreased glutathione level, total sulfhydryl content, and cellular antioxidant power. PCP treatment lowered the activity of antioxidant enzymes and inhibited enzymes of glucose metabolism. However, prior incubation with catechin attenuated the PCP-induced changes in all these parameters in a catechin concentration-dependent manner. Scanning electron microscopy of erythrocytes confirmed these biochemical results. PCP treatment converted the normal discoidal erythrocytes to irregularly contracted cells, acanthocytes, and echinocytes but the presence of catechin inhibited these morphological changes and erythrocytes retained their biconcave shape to a large extent. Genotoxicity was studied in human lymphocytes by single-cell gel electrophoresis (comet assay). It showed strand breaks and longer comet tail length in PCP alone treated cells. The comet tail length was reduced in the catechin +PCP-treated lymphocytes showing that catechin protected cells from PCP-induced DNA damage. These results show that catechin protects human blood cells against PCP-induced oxidative damage.