Biomarkeri oksidativnog stresa u jetri pacova subakutno tretiranih diazinonom

Abstract

Diazinon (DZN) je organofosfatni pesticid koji se široko koristio u poljoprivredi. Svoje toksične efekte ostvaruje inhibicijom acetilholinesteraze (AChE) i indukovanjem oksidativnog stresa. U ovoj studiji, Wistar pacovi su oralno tretirani diazinonom (55 mg/kg) tokom četiri različita perioda: 7, 14, 21 i 28 dana. Posle svakog vremenskog perioda uzorkovano je tkivo jetre za ispitivanje enzimskih i neenzimskih biomarkera oksidativnog stresa. Analizirane su aktivnosti enzima AChE, superoksid-dizmutaze 1 (SOD1), katalaze (CAT), mijeloperoksidaze (MPO) i paraoksonaze 1 (PON1). Stepen oštećenja ćelijske membrane i proteina praćen je određivanjem koncentracije malondialdehida (MDA) i karbonilnih grupa (CO). Najveća inhibicija AChE zabeležena je između 7. (p<0,01) i 14. dana (p<0,05), a najveća inhibicija SOD1 između 14. (p<0,01) i 21. dana (p<0,05) tretmana u odnosu na kotrolu. Aktivnost CAT je bila značajno povećana (p<0,001) u sva četiri vremenska perioda. Aktivnost MPO raste 14., 21. (p<0,01) i 28. dana (p<0,05), dok su markeri oštećenja proteina povećani 21. i 28. dana tretmana (p<0,001). U sva četiri posmatrana vremenska perioda, postoji veoma jaka korelacija između aktivnosti AChE i MDA, kao i između AChE i PON1. Takođe, veoma jaka korelacija je zabeležena 14. dana između aktivnosti AChE i MPO, a 7. dana između AChE i CAT aktivnosti. Prolongirana primena DZN dovodi do izraženog oksidativnog stresa u jetri. Biomarkeri oksidativnog stresa se nalaze u jakoj korelaciji sa holinergičkim mehanizmom delovanja DZN, odnosno inhibicijom AChE i parametrima inflamacije.

Diazinon (DZN) is an organophosphate pesticide widely used in agriculture. It achieves its toxic effects by inhibiting acetylcholinesterase (AChE) and inducing oxidative stress. In this study, Wistar rats were treated orally with diazinon (55 mg/kg) for four different periods: 7, 14, 21, and 28 days. After each period, liver tissue was examined for enzymatic and non-enzymatic biomarkers of oxidative stress. The activities of AChE, superoxide dismutase 1 (SOD1), catalase (CAT), myeloperoxidase (MPO), and paraoxonase 1 (PON1) were analyzed. The degree of cell membrane and protein damage was monitored by determining the concentration of malondialdehyde (MDA) and carbonyl groups (CO). The greatest inhibition of AChE was observed between days 7 (p<0.01) and 14 (p<0.05), and the greatest inhibition of SOD1 between days 14 (p<0.01) and 21 (p<0.05) of treatment compared with control. CAT activity was significantly increased (p<0.001) in all four time periods. MPO activity increased on days 14, 21 (p<0.01) and 28 (p<0.05), whereas protein damage markers increased on days 21 and 28 of treatment (p<0.001). In all four observed time periods, there is a very strong correlation between AChE and MDA activity and between AChE and PON1. In addition, a very strong correlation was observed between AChE and MPO activity on day 14 and between AChE and CAT activity on day 7. Prolonged administration of DZN leads to pronounced oxidative stress in the liver. Oxidative stress biomarkers correlate strongly with the cholinergic mechanism of action of DZN, i.e., AChE inhibition and inflammatory parameters.