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Pdf (428.57 K) Egyptian Journal of Occupational Medicine, 2020; 44 (2) : 663 - 678 Oxidative DNA Damage Due to Occupational Exposure to Polycyclic Aromatic Hydrocarbons among Coal Tar Workers BY Shehata RA1, Helal SF1, Rashed LA2 and Rakha AM1 1Department of Occupational and Environmental Medicine, 2Department of Biochemistry, Faculty of Medicine, Cairo University. Corresponding author: Shehata RA: [email protected] Abstract Introduction: Polycyclic Aromatic hydrocarbons (PAHs) are compounds of occupational and environmental concern as they are toxic, carcinogenic, mutagenic, and/or teratogenic (causing birth defects). Occupational exposure during coal tar distillation process may cause numerous symptoms and diseases through generation of coal tar fumes that contain principally polycyclic aromatic hydrocarbons (PAHs). Benzopyrene is the most important member of those PAHs. Aim of work: To assess the health effect due to occupational exposure to PAHs; specifically their effects on DNA damage and oxidative stress status among coal tar workers. Materials and methods: A comparative cross sectional study was carried out on two groups, exposed and unexposed group. The exposed group was consisted of 40 coal tar workers and the unexposed group was also composed of 40 matched individuals not occupationally exposed to coal tar .All examined individuals were subjected to full occupational history and estimation of 1-hydroxypyrene (1-OHPyrene) level in urine, blood levels of Benzopyrene-DNA adducts, oxidative stress enzymes (superoxide dismutase and glutathione peroxidase), Cytochrome P450 and Complete blood picture. Results: There was a statistically significant higher urinary level of 1-OHPyrene and higher blood levels of benzopyrene DNA adducts at the exposed workers compared to unexposed ones, in the other hand there were significant reduction in serum levels of superoxide dismutase and glutathione peroxidase among workers exposed to PAHs compared to the unexposed group that might be consumed by oxidative stress status. Conclusion: Oxidative stress and DNA damage might be one of the pathogenesis that may cause adverse health effects associated with occupational exposure to PAHs among coal tar workers. Keywords: Coal tar production, Polycyclic aromatic hydrocarbons, 1-Hydroxypyrene, DNA adducts and Oxidative stress enzymes. 664 Shehata et al. Introduction Individual PAHs such as benzo[a] Polycyclic aromatic hydrocarbons pyrene and mixtures containing various (PAHs) are a group of chemical PAHs have been determined to be compounds that is formed whenever carcinogenic to humans and animals. anything with a carbon base is burned, The route of exposure can influence from wood and gasoline to cigarettes and the site of tumour induction although meat. PAHs are also in automobile tires tumours can be present in other and coal tar production which involves locations such as lung cancers after heating of carbon-based materials. PAHs are of environmental concern because dermal exposures ( CRCE, 2008). they are toxic, mutagenic, teratogenic Benzo[a]pyrene and other polycyclic (causing birth defects) and harmful to aromatic hydrocarbons (PAHs) execute aquatic life, and even they are probable their carcinogenicity via formation human carcinogens (Abdel-Shafy and of benzopyrene DNA adducts. Mansour , 2016). Quantitation of DNA adducts in human Studies of coke-oven pitch tissues has been achieved with highly detected that coal tar pitch contained aromatic hydrocarbon ring systems sensitive techniques based on adduct including benzo[a]pyrene (BaP) and radio-labelling, antisera specific for benzo[e] pyrene, benzofluoranthenes, DNA adducts. Studies were used to perylene and heterocyclic compounds. correlate DNA adducts with cancer Several studies showed evidence risk, and elucidation of opportunities of carcinogenicity in human from to reduce human DNA adduct levels occupational exposures during paving formation (Poirier, 2004). and roofing with coal tar pitch (Betts, 2000). Benzopyerene DNA adduct was Benzo[a]pyrene (BaP) is one of the observed in different cell types. The most toxic components of the PAHs. levels were similar in various cell BaP rarely exists in the air on its own types; that suggested that DNA in many but rather is associated with a large human tissues is continuously damaged number of other PAHs present in both from known exposure of humans to vapour phase or as particles. In some BaP and other PAH. Moreover, DNA regulatory settings, BaP is used as an indicator for this group of chemically adducts formed from BaP are persistent diverse air pollutants ( Alexandrov et in lung and brain specifically ( Boysen al.,2010). and Hecht, 2003). Oxidative DNA and Occupational Exposure to Polycyclic Aromatic Hydrocarbons 665 Free radicals and reactive oxygen Study sample: The studied species (ROS) that are generated during population included 80 subjects divided metabolism of polycyclic aromatic into 40 exposed and 40 unexposed hydrocarbons (PAHs) can cause severe male workers. The exposed workers damage to cellular macromolecules (e.g. were taken from the section of coal DNA, proteins, and lipids) via oxidative tar distillation of coal manufacture stress which have been reported to be at Helwan district, with duration of involved in the development of diseases employment from 5 to 35 years. These including cardiovascular, diabetes, and workers were occupationally exposed cancers for adult and birth defects for to dust and fumes produced during fetus (Patri et al., 2010). coal tar distillation at 8 hours day shift for 5 days/week, the place is poorly Biochemical estimation of radical ventilated or illuminated with lack of scavenging antioxidant enzymes, like good housing .The other 40 unexposed superoxide dismutase and glutathione males were selected from the outpatient peroxidase, gives information on the clinics in Kasr Al-Ainy hospital with possible imbalance between oxidant no history of occupational exposure and antioxidant systems resulting on to coal tar dust or fumes. Both groups oxidative stress status responsible were matched in age, sex, smoking for the developing DNA damage and and socioeconomic standard with the carcinogenic mechanism (Risom et al., exposed group. Inclusion criteria: 2005). male workers exposed to polyaromatic Aim of the work hydrocarbons in their occupation for more than 5 years. Exclusion criteria The aim of this work is to assess involved cases with history of any type the health effects due to occupational of cancer and moderate to heavy smokers exposure to PAHs; specifically their (Smoking Index=20- 40 pack\year) (as effects on DNA damage and oxidative tobacco smoke is a major source of stress status among coal tar workers. human exposure to polycyclic aromatic Materials and methods hydrocarbons (PAHs) and increase Study design: Comparative cross cancer risk( Yan et al., 2006). Also presence of cancer have been reported sectional study. to be associated by oxidative damage to Place and duration of the study: biological molecules including DNA at The study was conducted at section of adults (Patri et al., 2010)). So smoking coal tar distillation of coal manufacture or cancer might affect the results of the at Helwan district, from October 2015 investigations of the present study. to June 2016. 666 Shehata et al. Study Methods: Superoxide dismutase The studied groups were subjected (SOD) activity was measured to the following: spectrophotometrically. Briefly, xanthine–xanthine oxidase was I- Questionnaire: Specially designed utilized to generate a superoxide flux. questionnaire was used included: Reduction of nitrobluetetrazolium medical, present, past and family (NBT) by superoxide anion to blue histories with emphasis on general formazon was determined at 560 toxic and respiratory manifestations. nm. One unit of enzyme activity was II-Clinical examination for: defined as the amount of protein causing • Vital signs: pulse, blood pressure, 50% inhibition in NBT reduction by respiratory rate and temperature. superoxide (Rekhadevi et al., 2009). Glutathione peroxidase (GPx) • Skin examination: redness, dryness, activity in serum was measured cracks, itching marks and blisters. spectrophotometrically. The enzyme • Chest examination: breath sounds, reaction was initiated by the addition wheezes and crepitations. of hydrogen peroxide (H2O2) to the III-Laboratory investigations: reaction medium and the rate of 1) Collection of blood samples About nicotinamide adenine dinucleotide 10 ml of venous blood was collected phosphate (NADPH) oxidation was once from each individual in the followed at 340 nm. The amount of examined groups using K EDTA enzyme that oxidizes 1 µmol NADPH 2 per minute was considered to be one plastic tubes. unit (Rekhadevi et al., 2009). Sampling process: Cytochrome P450 (CYP450) in a) Antioxidant enzyme assays serum activity was measured using (Superoxide dismutase and ELISA method. This assay employs Glutathione peroxidase) the quantitative sandwich enzyme A total of 1.5 ml of the total blood immunoassay technique. Antibody collected was processed to obtain specific for Cytochrome P450 has been pre-coated onto a microplate. Standards serum for biochemical analysis. Protein and samples are pipetted into the wells determination was done in all samples and any Cytochrome P450 present is (Rekhadevi et al., 2009). Oxidative DNA and Occupational Exposure to Polycyclic Aromatic Hydrocarbons 667 bound by the immobilized antibody. 2) Collection of urine samples: Urine After removing any unbound substances,
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