Diclofop-methyl: A phenoxy propionate herbicide with multiple toxic effects in mouse embyro fibroblast (NIH/3T3) cell line

Year 2017, Volume: 21 Issue: 4, 992 - 997, 01.12.2017

Müzeyyen Çeliksöz Bahar Ulus Ezgi Öztaş Gül Özhan

Abstract

Diclofop-methyl is a selective post-emergence graminicide from

the phenoxy propionate group of herbicides to be developed

for control of wild oats, millets, and other annual grass weeds.

Diclofop-methyl usage is limited in various grass weed species

due to its toxic effect and exposure risks. However, total annual

usage of is approximately 750.000 pounds in United States,

and more in Asia. Therefore, we aimed to investigate diclofopmethyl’s

toxic potentials in vitro and the following assays were

used; MTT assay for cytotoxicity, comet assay for genotoxicity,

generation of reactive oxygen species (ROS), malondialdehyde

(MDA) and glutathione (GSH) for the potential of oxidative

damage in mouse embryo fibroblast (NIH/3T3) cell line.

Diclofop-methyl was observed to reduce the cell viability

in a concentration manner and the half maximal inhibitory

concentration (IC50) value was 301.7 μM. Diclofop-methyl

caused DNA damage and oxidative stress at the concentrations

between 12.5-400 μM. Tail intensities were at the range of 1.24-

58.21% with increasing concentrations, which are approximately

≤ 1.63-fold of the negative control. Also, MDA levels were

increased ≥ ³11.4-fold of the negative control that denotes lipid

peroxidation was induced. However, there was no significant

increment in the ROS and GSH levels at all concentrations. In

view of the fact that ROS has not been detected, despite its level

of MDA proffers, the idea that oxidative damage may have been

caused by other mechanisms. Our results indicate that diclofopmethyl

was cytotoxic, genotoxic and might have oxidative

damage potential in vitro conditions.

Keywords

Diclofop-methyl; reactive oxygen species; malondialdehyde; Comet assay

References

• 1. Abd-alrahman SH, Elhalwagy MEA, Kotb GA, Farid H, Farag AAG, Draz HM, Isa AM, Sabico S . Exposure to difenoconazole, diclofop-methyl alone and combination alters oxidative stress and biochemical parameters in albino rats. Int J Clin Exp Med 2014; 7: 3637–46. 2. Larsen K, Najle R, Lifschitz A, Mate ML, Lanusse C, Virkel GL. Effects of sublethal exposure to a glyphosate-based herbicide formulation on metabolic activities of different xenobioticmetabolizing enzymes in rats. Int J Toxicol 2014; 33: 307–18. 3. Unal F, Yüzbaşıoğlu D, Yılmaz S, Akıncı N, Aksoy H. Genotoxic effects of chlorophenoxy herbicide diclofop-methyl in mice in vivo and in human lymphocytes in vitro. Drug Chem Toxicol 2011; 34: 390-5. 4. Shimabukuro R, Hoffer B. Enantiomers of diclofop-methyl and their role in herbicide mechanism of action. Pestic Biochem Physiol 1995; 51: 66–82. 5. Hongming L, Xu L, Zhaojian G, Fan Y. Isolation of an aryloxyphenoxy propanoate (AOPP) herbicide-degrading strain Rhodococcus ruber JPL-2 and the cloning of a novel carboxylesterase gene (feh). Brazilian J 2015; 432: 425–32. 6. United States Environmental Protection Agency (U.S. EPA). Prevention, pesticides and toxic substances. EPA- 738-F-00-007 (7508C) R.E.D. FACTS diclofop-methyl. Washington, DC; 2000. 7. Ding H, Lu H, Lavoie M, Xie J, Li Y, Lv X. Unraveling the toxicity mechanisms of the herbicide diclofop-methyl in rice: modulation of the activity of key enzymes involved in citrate metabolism and induction of cell membrane anion channels. J Agric 2014; 62: 10654-60. 8. Ye J, Wang L, Zhang Z, Liu W. Enantioselective physiological effects of the herbicide diclofop on cyanobacterium Microcystis aeruginosa. Environ Sci 2013; 47: 3893–901. 9. Eliyahu D, Applebaum S, Rafaeli A. Moth sex-pheromone biosynthesis is inhibited by the herbicide diclofop. Pestic Biochem Physiol 2003; 77: 75–81. 10. Meyer KJ, Reif JS, Rao Veeramachaneni DN, Luben TJ, Mosley BS, Nuckols JR. Agricultural pesticide use and hypospadias in Eastern Arkansas. Environ Health Perspect 2006; 114: 1589– 95. 11. Methods Analytical and Health Effects. Diclofop-Methyl 1987; (March 1987): 1–2. 12. Abudayyak M, Öztaş E, Arici M, Özhan G. Investigation of the toxicity of bismuth oxide oxide nanoparticles in various cell lines. Chemosphere 2017; 169: 117-23. 13. Alpertunga B, Kara M, Abudayyak M. Effects of prochloraz on DNA damage, lipid peroxidation and antioxidant system in vitro. Toxicol Mech Methods 2014; 24: 268-75. 14. Speit G, Hartmann A. The comet assay (Single-Cell Gel Test) a sensitive genotoxicity test for the detection of DNA damage and repair. DNA Repair Protoc Eukaryot Syst 1999; 203-12. 15. Collins AR. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 2004; 26: 249–61. 16. Sandal S, Yilmaz B. Genotoxic effects of chlorpyrifos, cypermethrin, endosulfan and 2,4‐D on human peripheral lymphocytes cultured from smokers and nonsmokers. Environ Toxicol 2011; 26: 433-42. 17. DiTomaso J, Brown P, Stowe A, Linscott D. Effects of diclofop and diclofop-methyl on methyl on membrane potentials in roots of intact oat, maize, and pea seedlings. Plant 1991; 95: 1063–9. 18. Bolognesi C, Morasso G. Genotoxicity of pesticides: potential risk for consumers. Trends Food Sci Technol 2000; 11: 182–7. 19. Department of National Health and Welfare. Unpublished studies; summary of evaluations by the Food Directorate (1988). 20. Cañas N, Valero T, Villarroya M, Montell E. Chondroitin sulfate protects SH-SY5Y cells from oxidative stress by inducing heme oxygenase-1 via phosphatidylinositol 3-kinase/ Akt. J Pharmacol Exp Ther 2007; 323: 946–53.