Many compounds have the potential to harm pancreatic beta-cells; organochlorine pollutants belong to those compounds. In this work, we aimed to find markers of acute toxicity of p,p'-DDT exposure among proteins expressed in NES2Y human pancreatic beta-cells employing 2-D electrophoresis. We exposed NES2Y cells to a high concentration (150 μM, LC96 after 72 hours) of p,p'-DDT for 24 and 30 hours and determined proteins with changed expression using 2-D electrophoresis. We have found 22 proteins that changed their expression. They included proteins involved in ER stress (GRP78, and endoplasmin), mitochondrial proteins (GRP75, ECHM, IDH3A, NDUS1, and NDUS3), proteins involved in the maintenance of the cell morphology (EFHD2, TCPA, NDRG1, and ezrin), and some other proteins (HNRPF, HNRH1, K2C8, vimentin, PBDC1, EF2, PCNA, biliverdin reductase, G3BP1, FRIL, and HSP27). The proteins we have identified may serve as indicators of p,p'-DDT toxicity in beta-cells in future studies, including long-term exposure to environmentally relevant concentrations.
- MeSH
- 2D gelová elektroforéza MeSH
- beta-buňky cytologie účinky léků metabolismus MeSH
- biologické markery metabolismus MeSH
- buněčné linie MeSH
- DDT toxicita MeSH
- hmotnostní spektrometrie MeSH
- lidé MeSH
- proteomika metody MeSH
- regulace genové exprese účinky léků MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Persistent organochlorine pollutants (POPs) gradually accumulate in the human organism due to their presence in the environment. Some studies have described a correlation between the level of POPs in the human body and the incidence of diabetes, but we know little about the direct effect of POPs on pancreatic beta-cells. We exposed pancreatic beta-cells INS1E to non-lethal concentrations of p,p'-DDT (1,1'-(2,2,2-Trichloroethane-1,1-diyl)bis(4-chlorobenzene)) and p,p'-DDE (1,1'-(2,2-dichloroethene-1,1-diyl)bis(4-chlorobenzene)) for 1 month, and assessed changes in protein expression and the intracellular insulin level. 2-D electrophoresis revealed 6 proteins with changed expression in cells exposed to p,p'-DDT or p,p'-DDE. One of the detected proteins - vitamin D-binding protein (VDBP) - was upregulated in both cells exposed to p,p'-DDT, and cells exposed to p,p'-DDE. Both exposures to pollutants reduced the intracellular level of insulin mRNA, proinsulin, and insulin monomer; p,p'-DDT also slightly reduced the level of hexameric insulin. Overexpression of VDBP caused by the stable transfection of beta-cells with the gene for VDBP decreased both the proinsulin and hexameric insulin level in beta-cells similarly to the reduction detected in cells exposed to p,p'-DDT. Our data suggest that in the cells exposed to p,p'-DDT and p,p'-DDE, the increased VDBP protein level decreased the proinsulin expression in an unknown mechanism.
- MeSH
- beta-buňky účinky léků metabolismus MeSH
- buněčné linie MeSH
- DDT toxicita MeSH
- dichlordifenyldichlorethylen toxicita MeSH
- inzulin metabolismus MeSH
- krysa rodu rattus MeSH
- látky znečišťující životní prostředí toxicita MeSH
- protein vázající vitamin D metabolismus MeSH
- testy subchronické toxicity MeSH
- upregulace účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nano-scale zero-valent iron (nZVI) has been conceived for cost-efficient degradation of chlorinated pollutants in soil as an alternative to e.g permeable reactive barriers or excavation. Little is however known about its efficiency in degradation of the ubiquitous environmental pollutant DDT and its secondary effects on organisms. Here, two types of nZVI (type B made using precipitation with borohydride, and type T produced by gas phase reduction of iron oxides under H2) were compared for efficiency in degradation of DDT in water and in a historically (>45 years) contaminated soil (24 mg kg(-1) DDT). Further, the ecotoxicity of soil and water was tested on plants (barley and flax), earthworms (Eisenia fetida), ostracods (Heterocypris incongruens), and bacteria (Escherichia coli). Both types of nZVI effectively degraded DDT in water, but showed lower degradation of aged DDT in soil. Both types of nZVI had negative impact on the tested organisms, with nZVI-T giving least adverse effects. Negative effects were mostly due to oxidation of nZVI, resulting in O2 consumption and excess Fe(II) in water and soil.
- MeSH
- chemické látky znečišťující vodu chemie toxicita MeSH
- DDT chemie toxicita MeSH
- ekotoxikologie MeSH
- Escherichia coli účinky léků MeSH
- ječmen (rod) účinky léků MeSH
- korýši účinky léků MeSH
- kyslík metabolismus MeSH
- látky znečišťující půdu chemie toxicita MeSH
- len účinky léků MeSH
- Oligochaeta účinky léků MeSH
- půda MeSH
- regenerace a remediace životního prostředí MeSH
- železo chemie toxicita MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Altered gap junctional intercellular communication (GJIC) has been associated with chemical carcinogenesis, where both chemical tumor promoters and chemopreventive agents (CPAs) are known to conversely modulate GJIC. The aim of this study was to investigate whether attenuation of chemically inhibited GJIC represents a common outcome induced by different CPAs, which could be effectively evaluated using in vitro methods. Rat liver epithelial cells WB-F344 were pretreated with a CPA for either 30 min or 24 h, and then exposed to GJIC-inhibiting concentration of a selected tumor promoter or environmental toxicant [12-O-tetradecanoylphorbol-13-acetate (TPA), lindane, fluoranthene, 1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane (DDT), perfluorooctanoic acid (PFOA), or pentachlorophenol]. Out of nine CPAs tested, quercetin and silibinin elicited the most pronounced effects, preventing the dysregulation of GJIC by all the GJIC inhibitors, but DDT. Metformin and curcumin attenuated the effects of three GJIC inhibitors, whereas the other CPAs prevented the effects of two (diallyl sulfide, emodin) or one (indole-3-carbinol, thymoquinone) GJIC inhibitor. Significant attenuation of chemically induced inhibition of GJIC was observed in 27 (50%) out of 54 possible combinations of nine CPAs and six GJIC inhibitors. Our data demonstrate that in vitro evaluation of GJIC can be used as an effective screening tool for identification of chemicals with potential chemopreventive activity.
- MeSH
- antikarcinogenní látky farmakologie MeSH
- DDT toxicita MeSH
- epitelové buňky účinky léků MeSH
- fluoreny toxicita MeSH
- fluorokarbony toxicita MeSH
- hexachlorcyklohexan toxicita MeSH
- játra cytologie účinky léků MeSH
- kapryláty toxicita MeSH
- karcinogeny toxicita MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- kurkumin farmakologie MeSH
- metformin farmakologie MeSH
- mezerový spoj účinky léků metabolismus MeSH
- mezibuněčná komunikace účinky léků MeSH
- potkani inbrední F344 MeSH
- tetradekanoylforbolacetát toxicita MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Pollution of the environment represents one of less explored potential reasons for the worldwide epidemic of type 2 diabetes. One of the most prevalent organochlorine pollutants remains the pesticide DDT and its degradation product DDE. Despite some epidemiologic correlations between levels of DDT and DDE in human organism and the prevalence of diabetes, there is almost no information about the exact targets of these compounds inside pancreatic beta cells. To detect functional areas of pancreatic beta cells that could be affected by exposure to DDT and DDE, we analyzed changes in protein expression in the NES2Y human pancreatic beta cell line exposed to three sublethal concentrations (0.1 μM, 1 μM, 10 μM) of DDT and DDE for 1 month. Protein separation and identification was achieved using high-resolution 2D-electrophoresis, computer analysis and mass spectrometry. With these techniques, four proteins were found downregulated after exposure to 10 μM DDT: three cytoskeletal proteins (cytokeratin 8, cytokeratin 18 and actin) and one protein involved in glycolysis (alpha-enolase). Two proteins were downregulated after exposure to 10 μM DDE: cytokeratin 18 and heterogenous nuclear ribonucleoprotein H1 (HNRH1). These changes correlate with previously described effects of other stress conditions (e.g. exposure to palmitate, hyperglycemia, imidazoline derivative, and cytokines) on protein expression in pancreatic beta cells. We conclude that cytoskeletal proteins and their processing, glucose metabolism, and mRNA processing may represent targets affected by exposure to conditions hostile to pancreatic beta cells, including exposure to DDT and DDE.
- MeSH
- beta-buňky účinky léků metabolismus MeSH
- buněčné linie MeSH
- cytoskeletální proteiny metabolismus MeSH
- DDT toxicita MeSH
- dichlordifenyldichlorethylen toxicita MeSH
- glukosa metabolismus MeSH
- látky znečišťující životní prostředí toxicita MeSH
- lidé MeSH
- messenger RNA metabolismus MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- Anopheles parazitologie růst a vývoj MeSH
- antimalarika imunologie škodlivé účinky terapeutické užití MeSH
- DDT dějiny škodlivé účinky toxicita MeSH
- epidemiologie trendy MeSH
- lidé MeSH
- malárie diagnóza etiologie terapie MeSH
- Plasmodium parazitologie růst a vývoj MeSH
- vakcína proti malárii aplikace a dávkování imunologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
DDT je organochlorový insekticid, kdysi intenzivně používaný v celém světě v zemědělství a při kontrole hmyzu přenášejícího malárii a tyfus. V roce 1972 bylo zakázáno používání DDT ve Spojených státech a podobně i v některých dalších zemích. V mnoha rozvojových zemích je DDT i nadále používán při regulaci komárů přenášejících malárii. DDT je považován za perzistentní organický polutant přetrvávající dlouhodobě v životním prostředí a v živočiších, schopný šíření na velké vzdálenosti. DDT je považován Světovou zdravotnickou organizací za pravděpodobný karcinogen. V důsledku stability a schopnosti akumulovat se v tucích byl nalezen v lidských tkáních a v současné době není na celé planetě jediný organismus, který by neobsahoval DDT. Ačkoliv existují přesvědčivé experimentální důkazy o karcinogenitě DDT a jeho hlavních metabolitů DDE a DDD, epidemiologické studie jsou neprůkazné a převažují negativní výsledky. Možný příspěvek DDT ke zvýšení rizika různých druhů rakoviny a jeho možného negativního účinku na endokrinní systémy si vyžádá ještě další studie.
DDT is an organic chlorinated insecticide, formerly intensively used throughout the world in agriculture and in the control of insects transmitting malaria and typhus. In 1972, the use of DDT was prohibited in the USA and similarly also in some other countries. In many developing countries, DDT is ever being used in the control of mosquitoes transmitting malaria. DDT is considered as a persisting organic pollutant that shows its long-term presence in the environment and in animals that can be propagated to long distances. DDT is considered as probably carcinogenic substance by the World Health Organization. Due to its stability and ability to be accumulated in lipids, it was found in human typhus, and there is currently no organism throughout the planet that would not contain DDT. In spite of existing convincing experimental demonstrations of the carcinogenicity of DDT and of its main metabolites DDE and DDD, epidemiological studies are not demonstrable, with a prevalence of negative results. Further studies will be necessary concerning possible contributions of DDT to enhancing the risk of different types of cancer diseases and its possible negative effects on the endocrine system.
