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Title: Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation

Abstract

Exposure to benzene, toluene and xylene in the human population may pose a health risk. We tested a working hypothesis that these test chemicals cause cellular toxicity to a non-target organism, Drosophila melanogaster. Third instar larvae of D. melanogaster transgenic for hsp70, hsp83 and hsp26 and Oregon R{sup +} strain were exposed to 1.0-100.0 mM benzene, toluene and xylene for 2-48 h to examine the heat shock proteins (hsps), ROS generation, anti-oxidant stress markers and developmental end points. The test chemicals elicited a concentration- and time-dependent significant (p < 0.01) induction of the hsps in the exposed organism in the order of hsp70 > hsp83 {>=} hsp26 as evident by {beta}-galactosidase activity after 24 h. RT-PCR amplification studies in Oregon R{sup +} larvae revealed a similar induction pattern of these genes along with hsp60 in the order of hsp70 > hsp60 > hsp26 {>=} hsp83. Under similar experimental conditions, a significant induction of ROS generation and oxidative stress markers viz. superoxide dismutase, catalase, glutathione S-transferase, thioredoxin reductase, glutathione, malondialdehyde and protein carbonyl content was observed. Sub-organismal response was propagated towards organismal response i.e., a delay in the emergence of flies and their reproductive performance. While hsp70 was predominantly induced inmore » the organism till 24 h of treatment with the test chemicals, a significant or insignificant regression of Hsp70 after 48 h was concurrent with a significant induction (p < 0.01) of hsp60 > hsp83 {>=} hsp26 in comparison to the former. A significant positive correlation was observed between ROS generation and these hsps in the exposed organism till 24 h and a negative correlation between ROS generation and hsp70 in them after 48 h indicating a modulatory role of ROS in the induction of hsps. The study suggests that among the tested hsps, hsp70 may be used as an early bioindicator of cellular toxicity against benzene, toluene and xylene and D. melanogaster as an alternative animal model for screening the risk posed by environmental chemicals.« less

Authors:
 [1];  [2];  [3];  [4];  [1];  [5]
  1. Embryotoxicology Section, Indian Institute of Toxicology Research, P.O. Box No. 80, Mahatma Gandhi Marg, Lucknow 226 001, Uttar Pradesh (India)
  2. Analytical Chemistry Section, Indian Institute of Toxicology Research, P.O. Box No. 80, Mahatma Gandhi Marg, Lucknow 226 001, Uttar Pradesh (India)
  3. Epidemiology Section, Indian Institute of Toxicology Research, P.O. Box No. 80, Mahatma Gandhi Marg, Lucknow 226 001, Uttar Pradesh (India)
  4. (India)
  5. Embryotoxicology Section, Indian Institute of Toxicology Research, P.O. Box No. 80, Mahatma Gandhi Marg, Lucknow 226 001, Uttar Pradesh (India), E-mail: dkarchowdhuri@rediffmail.com
Publication Date:
OSTI Identifier:
21182750
Resource Type:
Journal Article
Resource Relation:
Journal Name: Toxicology and Applied Pharmacology; Journal Volume: 235; Journal Issue: 2; Other Information: DOI: 10.1016/j.taap.2008.12.002; PII: S0041-008X(08)00505-X; Copyright (c) 2008 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BENZENE; BIOLOGICAL INDICATORS; CARBONYLS; CATALASE; DROSOPHILA; GALACTOSIDASE; GENES; GLUTATHIONE; HEALTH HAZARDS; HEAT-SHOCK PROTEINS; LARVAE; OXIDATION; OXIDIZERS; POLYMERASE CHAIN REACTION; SUPEROXIDE DISMUTASE; TOLUENE; TOXICITY; XYLENES

Citation Formats

Singh, Mahendra Pratap, Reddy, M.M. Krishna., Mathur, N., Council of Scientific and Industrial Research, New Delhi, Saxena, D.K., and Chowdhuri, D. Kar. Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation. United States: N. p., 2009. Web. doi:10.1016/j.taap.2008.12.002.
Singh, Mahendra Pratap, Reddy, M.M. Krishna., Mathur, N., Council of Scientific and Industrial Research, New Delhi, Saxena, D.K., & Chowdhuri, D. Kar. Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation. United States. doi:10.1016/j.taap.2008.12.002.
Singh, Mahendra Pratap, Reddy, M.M. Krishna., Mathur, N., Council of Scientific and Industrial Research, New Delhi, Saxena, D.K., and Chowdhuri, D. Kar. 2009. "Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation". United States. doi:10.1016/j.taap.2008.12.002.
@article{osti_21182750,
title = {Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation},
author = {Singh, Mahendra Pratap and Reddy, M.M. Krishna. and Mathur, N. and Council of Scientific and Industrial Research, New Delhi and Saxena, D.K. and Chowdhuri, D. Kar},
abstractNote = {Exposure to benzene, toluene and xylene in the human population may pose a health risk. We tested a working hypothesis that these test chemicals cause cellular toxicity to a non-target organism, Drosophila melanogaster. Third instar larvae of D. melanogaster transgenic for hsp70, hsp83 and hsp26 and Oregon R{sup +} strain were exposed to 1.0-100.0 mM benzene, toluene and xylene for 2-48 h to examine the heat shock proteins (hsps), ROS generation, anti-oxidant stress markers and developmental end points. The test chemicals elicited a concentration- and time-dependent significant (p < 0.01) induction of the hsps in the exposed organism in the order of hsp70 > hsp83 {>=} hsp26 as evident by {beta}-galactosidase activity after 24 h. RT-PCR amplification studies in Oregon R{sup +} larvae revealed a similar induction pattern of these genes along with hsp60 in the order of hsp70 > hsp60 > hsp26 {>=} hsp83. Under similar experimental conditions, a significant induction of ROS generation and oxidative stress markers viz. superoxide dismutase, catalase, glutathione S-transferase, thioredoxin reductase, glutathione, malondialdehyde and protein carbonyl content was observed. Sub-organismal response was propagated towards organismal response i.e., a delay in the emergence of flies and their reproductive performance. While hsp70 was predominantly induced in the organism till 24 h of treatment with the test chemicals, a significant or insignificant regression of Hsp70 after 48 h was concurrent with a significant induction (p < 0.01) of hsp60 > hsp83 {>=} hsp26 in comparison to the former. A significant positive correlation was observed between ROS generation and these hsps in the exposed organism till 24 h and a negative correlation between ROS generation and hsp70 in them after 48 h indicating a modulatory role of ROS in the induction of hsps. The study suggests that among the tested hsps, hsp70 may be used as an early bioindicator of cellular toxicity against benzene, toluene and xylene and D. melanogaster as an alternative animal model for screening the risk posed by environmental chemicals.},
doi = {10.1016/j.taap.2008.12.002},
journal = {Toxicology and Applied Pharmacology},
number = 2,
volume = 235,
place = {United States},
year = 2009,
month = 3
}
  • Monocyclic aromatic hydrocarbons (MAHs) such as benzene, toluene and xylene are being extensively used for various industrial and household purposes. Exposure to these hydrocarbons, occupationally or non-occupationally, is harmful to organisms including human. Several studies tested for toxicity of benzene, toluene and xylene, and interestingly, only a few studies looked into the attenuation. We used Drosophila model to test the genotoxic and apoptotic potential of these compounds and subsequently evaluated the efficiency of two phytochemicals, namely, quercetin and curcumin in attenuating test chemical induced toxicity. We exposed third instar larvae of wild type Drosophila melanogaster (Oregon R{sup +}) to 1.0-100.0more » mM benzene, toluene or xylene, individually, for 12, 24 and 48 h and examined their apoptotic and genotoxic potential. We observed significantly (P < 0.001) increased apoptotic markers and genotoxicity in a concentration- and time-dependent manner in organisms exposed to benzene, toluene or xylene. We also observed significantly (P < 0.001) increased cytochrome P450 activity in larvae exposed to test chemicals and this was significantly reduced in the presence of 3',4'-dimethoxyflavone, a known Aryl hydrocarbon receptor (AhR) blocker. Interestingly, we observed a significant reduction in cytochrome P450 activity, GST levels, oxidative stress parameters, genotoxic and apoptotic endpoints when organisms were exposed simultaneously to test chemical along with quercetin or curcumin. The study further suggests the suitability of D. melanogaster as an alternate animal model for toxicological studies involving benzene, toluene and xylene and its potential in studying the protective role(s) of phytochemicals.« less
  • To investigate the use of stress proteins hsp60 and hsp70 as sublethal biomarkers for contaminant exposure in sediments, two infaunal (Ampelisca abdita, estuarine; Rhepoxynius abronius, marine) and one epifaunal (Hyalella azteca, freshwater) amphipod species were exposed for 24 h to solutions of the heavy metal cadmium, the pesticides diazinon and dieldrin, and the polycyclic aromatic hydrocarbon fluoranthene. All three species are routinely used in standard sediment toxicity tests. Analysis of hsp60 and hsp70 was performed using western blotting techniques with subsequent comparative quantification by densitometry. Results demonstrated compound and species-specific induction of stress protein synthesis. Whereas one member of themore » hsp70 protein family showed the most sensitive response to xenobiotic compounds in H. azteca, several members of the hsp60 protein family were the main proteins induced in A. abdita and R. abronius. Sensitivity of the detected stress protein response was highest in H. azteca with significant effects at concentrations 110-, 50-, >1,000-, and >1-fold lower than LC50 values for cadmium, diazinon, dieldrin, and fluoranthene, respectively. The corresponding values were >5 (cadmium), 0.7 (diazinon), >1 (dieldrin), and 2.9 (fluoranthene) for A. abdita, and >2 (cadmium), 3.1 (diazinon), > 100 (dieldrin), and >2.9 (fluoranthene) for R. abronius.« less
  • Due to the intensive commercial application of silver nanoparticles (Ag NPs), risk assessment of this nanoparticle is of great importance. Our previous in vitro study demonstrated that Ag NPs caused DNA damage and apoptosis in mouse embryonic stem cells and fibroblasts. However, toxicity of Ag NPs in vivo is largely lacking. This study was undertaken to examine the toxic effects of well-characterized polysaccharide coated 10 nm Ag NPs on heat shock stress, oxidative stress, DNA damage and apoptosis in Drosophila melanogaster. Third instar larvae of D. melanogaster were fed a diet of standard cornmeal media mixed with Ag NPs atmore » the concentrations of 50 and 100 mug/ml for 24 and 48 h. Ag NPs up-regulated the expression of heat shock protein 70 and induced oxidative stress in D. melanogaster. Malondialdehyde level, an end product of lipid peroxidation was significantly higher while antioxidant glutathione content was significantly lower in Ag NPs exposed organisms. Activities of antioxidant enzyme superoxide dismutase and catalase were also significantly higher in the organisms exposed to Ag NPs. Furthermore, Ag NPs up-regulated the cell cycle checkpoint p53 and cell signaling protein p38 that are involved in the DNA damage repair pathway. Moreover, activities of caspase-3 and caspase-9, markers of apoptosis were significantly higher in Ag NPs exposed organisms. The results indicate that Ag NPs in D. melanogaster induce heat shock stress, oxidative stress, DNA damage and apoptosis. This study suggests that the organism is stressed and thus warrants more careful assessment of Ag NPs using in vivo models to determine if chronic exposure presents developmental and reproductive toxicity.« less