Cytotoxic Effect of Clethodim Exposure on Liver Cells&

Year 2024, Volume: 11 Issue: 2, 454 - 462, 30.04.2024

Ergün Ergenekon , Ömer Erman , Nihal Şimşek Özek

https://doi.org/10.30910/turkjans.1442321

Abstract

Clethodim, a herbicide group pesticide, is classified as a toxicity class II agent. The development toxicity, immunotoxicity, neurotoxicity and reproductive toxicity of this herbicide have been demonstrated in zebrafish and rat. There are no in vitro studies on the hepatoxic effects of clethodim. In this study, for the first time, clethodim was tested with liver cell line models. THLE-2 and B129 cells were treated with a wide range of clethodim for 96 hours. After that, its hepatoxic effects were evaluated by MTT and SRB analyses. Both assays revealed that the cellular proliferation was strikingly decreased with the dose dependent manner. Especially, 1000 µg/ml dose of the clethodim exposure supressed the cellular proliferation at a %90 and %75 for B129 and THLE-2 cells. Moreover, a significant reduction was acquired in both cells number and volume with the high doses of this cyclohexanedione class herbicide. These cytotoxic impacts can be strongly related to the herbicide elaved oxidative stress. In the light of our results, the long and high doses of clethodim exposure has a hepatoxic effect and its toxic target organ is be liver. Hence, further studies on the use of clethodim-based herbicides are urgently needed.

Keywords

Clethodim, human liver cell, mouse liver cell, MTT, SRB assay.

Supporting Institution

Atatürk University Scientific Research Projects Coordination Unit

Project Number

BAP, No: FYL-2023-12045

Thanks

We would be very grateful to is Anatolia High Technology Application and Research Center (DAYTAM) of Atatürk University for supporting lab facility.

Klethodim’e Maruz Kalmanın Karaciğer Hücreleri Üzerindeki Sitotoksik Etkisi&

Abstract

Herbisit grubu pestisit olan kletodim, toksisite sınıfı II ajan olarak sınıflandırılır. Bu herbisitin gelişim toksisitesi, immünotoksisitesi, nörotoksisitesi ve üreme toksisitesi zebra balığı ve sıçanlarda gösterilmiştir. Kletodim'in hepatoksik etkilerine ilişkin in vitro çalışma bulunmamaktadır. Bu çalışmada ilk kez kletodim karaciğer hücre hattı modelleriyle test edildi. THLE-2 ve B129 hücreleri, 96 saat boyunca geniş bir aralıktaki klethodim ile muamele edildi. Daha sonra hepatoksik etkileri MTT ve SRB testleri ile değerlendirildi. Her iki analiz de hücresel proliferasyonun doza bağlı olarak çarpıcı biçimde azaldığını ortaya çıkardı. Özellikle 1000 µg/ml dozunda kletodim maruziyetinin B129 ve THLE-2 hücrelerinde hücresel poliferasyonu %90 ve %75 oranında baskıladığı görüldü. Ayrıca sikloheksandion sınıfı herbisitin yüksek dozları ile hem hücre sayısında hem de hacminde önemli bir azalma elde edilmiştir. Bu sitotoksik etkiler, herbisitlerin neden olduğu oksidatif stresle güçlü bir şekilde ilişkili olabilir. Sonuçlarımız ışığında, kletodim'e uzun ve yüksek dozda maruz kalmanın hepatoksik etkiye sahip olduğu ve toksik hedef organının karaciğer olduğu görülmüştür. Bu nedenle, kletodim bazlı herbisit kullanımı hakkınca acil olarak daha fazla çalışmaya ihtiyaç vardır.

Keywords

Kletodim, insan karaciğer hücresi, fare karaciğer hücresi, MTT, SRB testleri.

Supporting Institution

Atatürk Üniversitesi Bilimsel Araştırmalar ve Koordinasyon Birimi

Project Number

BAP, No: FYL-2023-12045

References

• Ankit, Saha, L., Kishor, V., and Bauddh, K. 2020. Impacts of synthetic pesticides on soil health and non-targeted flora and fauna. Ecological and practical applications for sustainable agriculture, 65-88.
• Bahuguna, A., Khan, I., Bajpai, V. K., and Kang, S. C. 2017. MTT assay to evaluate the cytotoxic potential of a drug. ||| Bangladesh Journal of Pharmacology |||, 12(2): 115-118.
• Brêda-Alves, F., Militão, F. P., de Alvarenga, B. F., Miranda, P. F., de Oliveira Fernandes, V., Cordeiro-Araújo, M. K., and Chia, M. A. 2020. Clethodim (herbicide) alters the growth and toxins content of Microcystis aeruginosa and Raphidiopsis raciborskii. Chemosphere, 243: 125-318.
• Çakır, M., and Erden, Y. 2019. MCF-7 ve PC-3 hücre hatlarında, hücre canlılığı ve dna hasarı üzerine trpv4 antagonisti rn 1734'ün etkileri. Bozok Tıp Dergisi.
• Damalas, C.A., and Koutroubas, S. D. 2016. Farmers’ exposure to pesticides: toxicity types and ways of prevention. In Toxics, 4(1).
• Dcunha, R., Kumari, S., Najar, M. A., Aravind, A., Suvarna, K. S., Hanumappa, A., Mutalik, S. P., Mutalik, S., Kalthur, S. G., Rajanikant, G. K., Siddiqui, S., Alrumman, S., Alamri, S. A. M., Raghu, S. V., Adiga, S. K., Kannan, N., Thottethodi Subrahmanya, K. P., and Kalthur, G. 2023. High doses of clethodim-based herbicide GrassOut Max poses reproductive hazard by affecting male reproductive function and early embryogenesis in Swiss albino mice. Chemosphere, 336.
• Gavrilescu, M., Demnerová, K., Aamand, J., Agathos, S., and Fava, F. 2015. Emerging pollutants in the environment: Present and future challenges in biomonitoring, ecological risks and bioremediation. New Biotechnology, 32(1): 147–156.
• Ghasemi, M., Turnbull, T., Sebastian, S., and Kempson, I. 2021. The MTT assay: utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. International journal of molecular sciences, 22(23): 12827.
• Holen, E., Kausland, A., and Skjærven, K. 2010. Embryonic stem cells isolated from Atlantic cod (Gadus morhua) and the developmental expression of a stage-specific transcription factor ac-Pou2. Fish Physiology and Biochemistry, 36(4): 1029–1039.
• Hong, Z., Faqir, Y., Kalhoro, G. M., Kalhoro, M. T., Kaleri, A. R., Murad, A., and Adeel, M. 2023. Hazardous ımpacts of Clopyralid, Benazolin, and Clethodim on seed germination and seedling of oil rapeseed (Brassica napus). Polish Journal of Environmental Studies, 32(3): 2623-2636.
• Juan, C. A., Pérez de la Lastra, J. M., Plou, F. J., and Pérez-Lebeña, E. 2021. The chemistry of reactive oxygen species (ROS) revisited: outlining their role in biological macromolecules (DNA, lipids and proteins) and induced pathologies. International journal of molecular sciences, 22(9): 46-42.
• Keepers, Y. P., Pizao, P. E., Peters, G. J., van Ark-Otte, J., Winograd, B., and Pinedo, H. M. 1991. Comparison of the sulforhodamine B protein and tetrazolium (MTT) assays for in vitro chemosensitivity testing. European Journal of Cancer and Clinical Oncology, 27(7): 897–900.
• Khozimy, A. M., Abuzeid, M. A., Hamada, S.H.E., and El-Danasoury, H.M. 2021. Sub-acute toxicity of two herbicides halosulfuron-methyl and Clethodim on biochemical and hematological parameters in male albino rats. Journal of Applied Plant Protection, 10(1): 35-42.
• Kishor, D. R., Moses, S., & Kumar, M. 2020. Study of Important Insecticide Samples. Agriculture & Food: e-Newsletter, 5.
• Laetz, C. A., Baldwin, D. H., Collier, T. K., Hebert, V., Stark, J. D., and Scholz, N. L. 2009. The synergistic toxicity of pesticide mixtures: Implications for risk assessment and the conservation of endangered Pacific salmon. Environmental Health Perspectives, 117(3): 348–353.
• Martins, R. X., Carvalho, M., Maia, M. E., Flor, B., Souza, T., Rocha, T. L., and Farias, D. 2024. 2,4-D Herbicide-ınduced hepatotoxicity: Unveiling disrupted liver functions and associated biomarkers. Toxics, 12(1): 35.
• Mohamed, I. A., Omar, E. M., Tawfik, A. I., Amro, A. M. A., and Al Naggar, Y. 2023. Sublethal effects of herbicides clethodim, haloxyfop-P-methyl, and their mixture on honey bee health. Apidologie, 54(1).
• Pang, K., and Hu, J. 2020. Simultaneous analysis and dietary exposure risk assessment of fomesafen, clomazone, clethodim and its two metabolites in soybean ecosystem. International Journal of Environmental Research and Public Health, 17(6): 1951.
• Patil, P. B., Minteer, S. D., Mielke, A. A., Lewis, L. R., Casmaer, C. A., Barrientos, E. J., Ju, J. S., Smith, J. L., and Fisher, J. S. 2007. Malonyl coenzyme A affects insulin-stimulated glucose transport in myotubes. Archives of Physiology and Biochemistry, 113(1): 13–24.
• Saito, Y., Nishio, K., Ogawa, Y., Kimata, J., Kinumi, T., Yoshida, Y., Noguchi, N.,& Niki, E. 2006. Turning point in apoptosis/necrosis induced by hydrogen peroxide. Free radical research, 40(6), 619-630.
• Santos, K. R., Galter, I. N., Grecco, K. D., Silva, E. Z. M., Lascola, M. B., Dos Santos, S. N., and Matsumoto, S. T. 2023. Toxicogenetic, biochemical, and anatomical effects of the herbicide Clethodim on Allium cepa L. Environmental Science and Pollution Research, 30(60): 125388-125397.
• Skehan, P., Storeng, R., Scudiero, D., Monks, A., Mcmahon, J., Vistica, D., … Boyd, M. R. 1990. ARTICLES New Colorimetric Cytotoxicity Assay for Anticancer-Drug Screening. In J Natl Cancer Inst (Vol. 82). Retrieved from https://academic.oup.com/jnci/article-abstract/82/13/1107/970741.
• Vichai, V., and Kirtikara, K. 2006. Sulforhodamine B colorimetric assay for cytotoxicity screening. Nature Protocols, 1(3).
• Wang, H., Zhou, L., Meng, Z., Su, M., Zhang, S., Huang, P., Jiang, F., Liao, X., Cao, Z., and Lu, H. 2019. Clethodim exposure induced development toxicity and behaviour alteration in early stages of zebrafish life. Environmental Pollution, 255.
• Xiong, G., Zou, L., Deng, Y., Meng, Y., Liao, X., and Lu, H. 2019. Clethodim exposure induces developmental immunotoxicity and neurobehavioral dysfunction in zebrafish embryos. Fish and Shellfish Immunology, 86: 549–558.