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

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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 (0.05–2.5 mM) for 30 min followed by addition of 0.75 mM PCP and further incubation for 4 h 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.

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Abbreviations

ABTS:

2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)

AChE:

Acetylcholinesterase

AFR:

Ascorbate free radical

AMP:

Adenosine 5′-monophosphate

AO:

Antioxidant

AOs:

Antioxidants

AOPP:

Advanced oxidation protein products

ATP:

Adenosine 5′-triphosphate

ATPase:

Adenosine triphosphatase

CTN:

Catechin

CUPRAC:

Cupric reducing antioxidant capacity

DHE:

Dihydroethidium

DCFH-DA:

2,7-dichlorodihydrofluorescein diacetate

DPPH:

2,2-diphenyl-1-picrylhydrazyl

DTNB:

5,5′-dithiobisnitrobenzoic acid

EDTA:

Ethylenediaminetetraacetic acid

FRAP:

Ferric reducing antioxidant power

GSH:

Reduced glutathione

GSSG:

Oxidized glutathione

G6PD:

Glucose 6-phosphate dehydrogenase

GR:

Glutathione reductase

GST:

Glutathione-S-transferase

GPx:

Glutathione peroxidase

Hb:

Hemoglobin

H2O2 :

Hydrogen peroxide

LDH:

Lactate dehydrogenase

LPO:

Lipid peroxidation

NADP+ and NADPH:

Oxidized and reduced nicotinamide adenine dinucleotide phosphate

NAD+ and NADH:

Oxidized and reduced nicotinamide adenine dinucleotide

Na+K+ATPase:

Sodium potassium ATPase

PCP:

Pentachlorophenol

PK:

Pyruvate kinase

PMRS:

Plasma membrane redox system

RBC:

Red blood cells

RNS:

Reactive nitrogen species

ROS:

Reactive oxygen species

SH:

Sulfhydryl

SOD:

Cu-Zn superoxide dismutase

TBS:

Tris buffered saline

TR:

Thioredoxin reductase

Tris:

Tris(hydroxymethyl)aminomethane

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Acknowledgment

Financial support to the Department of Biochemistry from the UGC-SAP-DRS III, DST-PURSE II, and DBT-BUILDER programs is gratefully acknowledged. NM is the recipient of Senior Research Fellowship from CSIR, New Delhi.

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  1. Department of Biochemistry, Faculty of Life Sciences, Aligarh Muslim University, Aligarh, U. P., 202002, India

    Nikhil Maheshwari & Riaz Mahmood

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  1. Nikhil Maheshwari
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Maheshwari, N., Mahmood, R. Protective effect of catechin on pentachlorophenol-induced cytotoxicity and genotoxicity in isolated human blood cells. Environ Sci Pollut Res 27, 13826–13843 (2020). https://doi.org/10.1007/s11356-020-07969-0

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