hrvatski

Citotoksični, genotoksični i citogenetički učinci klorpirifosa na ljudske limfocite periferne krvi u uvjetima in vitro / Evaluation of the cytotoxic, genotoxic and cytogenetic effects of chlorpyrifos in human peripheral blood lymphocytes in vitro

Klorpirifos je jedan od najčešće korištenih organofosfornih insekticida. Zbog visoke lipofilnosti, molekula klorpirifosa lako ulazi u citoplazmu i oštećuje stanične makromolekule. U uvjetima in vitro istražili smo utjecaj klorpirifosa na apoptozu i nekrozu, oštećenja DNA te citogenetička oštećenja u ljudskim limfocitima periferne krvi, izlaganim 4 i 24 h koncentracijama koje odgovaraju: razini na kojoj je uočen štetan učinak, prihvatljivom dnevnom unosu te referentnoj razini izloženosti. Preživljenje stanica praćeno je primjenom testa s fluorescencijskim bojama. Primarna oštećenja DNA procijenjena su komet‐ testom, a citogenetička oštećenja mikronukleus‐ testom. Klorpirifos pri testiranim koncentracijama izaziva porast smrtnosti limfocita (najviše apoptozom), ovisno o koncentraciji i dužini izlaganja. Rezultati komet‐testa upućuju na koncentracijski ovisan porast razine primarnih oštećenja DNA. Pomoću mikronukleus‐testa utvrđena su značajna oštećenja kromosoma i diobenog vretena te poremećaji stanične kinetike i proliferacije limfocita. Citotoksični, genotoksični i citogenetički učinci klorpirifosa, uočeni čak i pri njegovim dopuštenim koncentracijama, pozivaju na povećani oprez, osobito jer neke namirnice mogu sadržavati visoke koncentracije ostataka ovog pesticida. Stoga je, radi smanjenja rizika od izloženosti klorpirifosu, potrebno nastaviti istraživanja s ciljem boljeg razjašnjenja mehanizama njegove toksičnosti. Chlorpyrifos is one of the most widely used organophosphate insecticides. Due to its lipophilicity, chlorpyrifos reaches the cytoplasm easily and damages cellular macromolecules. This in vitro study investigated the ability of chlorpyrifos to induce apoptosis/necrosis, DNA lesions and cytogenetic damage in human lymphocytes treated for 4 and 24 h with chlorpyrifos at doses equal to its observable effect level, acceptable daily intake, and reference exposure level. Cytotoxicity was tested using a viability assay with fluorescence dyes. Levels of primary DNA and cytogenetic damage were studied by comet assay and cytokinesis‐blocked micronucleus (CBMN) assay, respectively. Following treatment, lymphocyte viability decreased in a time‐ and concentration‐ dependent manner. Apoptosis dominated over necrosis. The levels of primary DNA damage were also concentration‐ dependent, but longer exposure resulted in slightly lower values, both of comet tail lengths and of tail DNA. The findings of CBMN assay indicate significant DNA damage, aneugenic potential, and disturbances in cell‐ cycle kinetics. The fact that chlorpyrifos was, even at concentrations tolerable for human exposure, able to induce significant cytotoxic, genotoxic and cytogenetic effects calls for particular attention, since some food items often contain high residual levels of this pesticide. In this sense, further studies aimed at clarifying the mechanisms involved in chlorpyrifos toxicity to a greater extent are advised. *Acknowledgement: This work was financially supported by the Project No. 8366 Organic Pollutants in Environment - Markers and Biomarkers of Toxicity (OPENTOX), funded by the Croatian Science Foundation.

toksičnost; oštećenja DNA; pesticid; ljudske krvne stanice

nije evidentirano

engleski

Evaluation of the cytotoxic, genotoxic and cytogenetic effects of chlorpyrifos in human peripheral blood lymphocytes in vitro

Klorpirifos je jedan od najčešće korištenih organofosfornih insekticida. Zbog visoke lipofilnosti, molekula klorpirifosa lako ulazi u citoplazmu i oštećuje stanične makromolekule. U uvjetima in vitro istražili smo utjecaj klorpirifosa na apoptozu i nekrozu, oštećenja DNA te citogenetička oštećenja u ljudskim limfocitima periferne krvi, izlaganim 4 i 24 h koncentracijama koje odgovaraju: razini na kojoj je uočen štetan učinak, prihvatljivom dnevnom unosu te referentnoj razini izloženosti. Preživljenje stanica praćeno je primjenom testa s fluorescencijskim bojama. Primarna oštećenja DNA procijenjena su komet‐ testom, a citogenetička oštećenja mikronukleus‐ testom. Klorpirifos pri testiranim koncentracijama izaziva porast smrtnosti limfocita (najviše apoptozom), ovisno o koncentraciji i dužini izlaganja. Rezultati komet‐testa upućuju na koncentracijski ovisan porast razine primarnih oštećenja DNA. Pomoću mikronukleus‐testa utvrđena su značajna oštećenja kromosoma i diobenog vretena te poremećaji stanične kinetike i proliferacije limfocita. Citotoksični, genotoksični i citogenetički učinci klorpirifosa, uočeni čak i pri njegovim dopuštenim koncentracijama, pozivaju na povećani oprez, osobito jer neke namirnice mogu sadržavati visoke koncentracije ostataka ovog pesticida. Stoga je, radi smanjenja rizika od izloženosti klorpirifosu, potrebno nastaviti istraživanja s ciljem boljeg razjašnjenja mehanizama njegove toksičnosti. Chlorpyrifos is one of the most widely used organophosphate insecticides. Due to its lipophilicity, chlorpyrifos reaches the cytoplasm easily and damages cellular macromolecules. This in vitro study investigated the ability of chlorpyrifos to induce apoptosis/necrosis, DNA lesions and cytogenetic damage in human lymphocytes treated for 4 and 24 h with chlorpyrifos at doses equal to its observable effect level, acceptable daily intake, and reference exposure level. Cytotoxicity was tested using a viability assay with fluorescence dyes. Levels of primary DNA and cytogenetic damage were studied by comet assay and cytokinesis‐blocked micronucleus (CBMN) assay, respectively. Following treatment, lymphocyte viability decreased in a time‐ and concentration‐ dependent manner. Apoptosis dominated over necrosis. The levels of primary DNA damage were also concentration‐ dependent, but longer exposure resulted in slightly lower values, both of comet tail lengths and of tail DNA. The findings of CBMN assay indicate significant DNA damage, aneugenic potential, and disturbances in cell‐ cycle kinetics. The fact that chlorpyrifos was, even at concentrations tolerable for human exposure, able to induce significant cytotoxic, genotoxic and cytogenetic effects calls for particular attention, since some food items often contain high residual levels of this pesticide. In this sense, further studies aimed at clarifying the mechanisms involved in chlorpyrifos toxicity to a greater extent are advised. *Acknowledgement: This work was financially supported by the Project No. 8366 Organic Pollutants in Environment - Markers and Biomarkers of Toxicity (OPENTOX), funded by the Croatian Science Foundation.

toxicity; DNA damage; pesticide; human blood cells

nije evidentirano