The Role of Flupyradifuron in Inducing Apoptosis in HMC3 Cells: An In Vitro Analysis

Abstract

Objective Flupyradifurone, a neonicotinoid insecticide, is widely applied in pest management. The present study aimed to elucidate the underlying mechanisms of flupyradifurone-induced toxicity in HMC3 microglial cells. Material and Method Cell viability was assessed by the MTT assay following exposure to flupyradifurone. In parallel, the expression of key apoptotic pathway genes, including MDM2, p53, and CASP9, as well as the apoptosisand cell viability–associated microRNA hsa-miR-21-5p, was evaluated using RT-PCR. Results The results demonstrated that flupyradifurone induced dose-dependent cell death in HMC3 cells. RT-PCR analysis revealed a significant downregulation of MDM2, miR-21-5p, and an upregulation of p53 and CASP9 after 24 h of treatment. Conclusion This study reveals that FDP causes cell death in HMC3 cells, and this process occurs through apoptosis.

Keywords

• Apoptosis
• Casp9
• flupyradifurone
• hsamiR-21-5p
• MDM2
• p53

References

1. 1. Zhong K, Meng Y, Wu J, Wei Y, Huang Y, Ma J, Lu H. Effect of flupyradifurone on zebrafish embryonic development. Environmental pollution 2021;285:117323.
2. 2. Strouhova A, Velisek J, Stara A. Selected neonicotinoids and associated risk for aquatic organisms. Veterinární medicína 2023;68(8):313.
3. 3. Wren M, Robson M, Buckley B. A novel biomonitoring method to detect pyrethroid metabolites in the saliva of occupationally exposed workers as a tool for risk assessment. Human and Ecological Risk Assessment: An International Journal 2024:30(3–4);269–288 doi: 10.1080/10807039.2024.2329625.
4. 4. Qiao Z, Li P, Tan J, Peng C, Zhang F, Zhang W, Jiang X. Oxidative stress and detoxification mechanisms of earthworms (Eisenia fetida) after exposure to flupyradifurone in a soil-earthworm system. Journal of Environmental Management 2022;322:115989.
5. 5. Lu Q, Ding H, Liu L, Xu L, Kuang H, Xu C, Guo L. Immunochromatographic assay for rapid detection of flupyradifurone in grape, blueberry, and tomato samples. Food Chemistry 2024;433:137328.
6. 6. Wu S-Y, Xing F, Sharma S, Wu K, Tyagi A, Liu Y, Zhao D, Deshpande RP, Shiozawa Y, Ahmed T. Nicotine promotes brain metastasis by polarizing microglia and suppressing innate immune function. Journal of Experimental Medicine 2020;217(8):e20191131.
7. 7. Resende RR, Adhikari A. Cholinergic receptor pathways involved in apoptosis, cell proliferation, and neuronal differentiation. Cell Communication and Signaling 2009;7:20.
8. 8. Blesa J, Trigo-Damas I, Quiroga-Varela A, Jackson-Lewis VR. Oxidative stress and Parkinson’s disease. Frontiers in neuroanatomy 2015;9:91.

9. 9. Nakamura T, Lipton SA. Cell death: protein misfolding and neurodegenerative diseases. Apoptosis 2009;14:455-68.
10. 10. Zhao J, Yu S, Zheng Y, Yang H, Zhang J. Oxidative modification and its implications for the neurodegeneration of Parkinson’s disease. Molecular neurobiology 2017;54:1404-18.
11. 11. Riss TL, Moravec RA, Niles AL, Duellman S, Benink HA, Worzella TJ, Minor L. Cell viability assays. In: Markossian S, Grossman A, Baskir H, et al. Assay Guidance Manual [Internet], Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences 2016; Available from: https:// www.ncbi.nlm.nih.gov/books/NBK144065/pdf/Bookshelf_ NBK144065.pdf
12. 12. Liu C, Tong Z, Tan J, Xin Z, Wang Z, Tian L. MicroRNA‑21‑5p targeting PDCD4 suppresses apoptosis via regulating the PI3K/AKT/FOXO1 signaling pathway in tongue squamous cell carcinoma. Experimental and therapeutic medicine 2019;18(5):3543-51.
13. 13. Tepebaşı MY, Öztürk Ö. miR-21, miR-221, and miR-222 upregulation in lung cancer promotes metastasis by reducing oxidative stress and apoptosis. Revista da Associação Médica Brasileira 2023;69:e20221688.
14. 14. Chakrabarti P, Carlson EA, Lucas HM, Melathopoulos AP, Sagili RR. Field rates of Sivanto™(flupyradifurone) and Transform®(- sulfoxaflor) increase oxidative stress and induce apoptosis in honeybees (Apis mellifera L.). Plos one 2020;15(5):e0233033.
15. 15. Xu L, Xu X, Wu X, Kuang H, Xu C. Sex-dependent environmental health risk analysis of flupyradifurone. Environmental science & technology 2022;56(3):1841-53.
16. 16. Şekeroğlu ZA, Aydın A, Yedier SK, Şekeroğlu V. Cytogenetic alterations induced by flupyradifurone, a new butenolide insecticide, in human lymphocytes. Toxicology and Industrial Health 2018;34(11):737-43.
17. 17. Zafar A, Khan MJ, Naeem A. MDM2-an indispensable player in tumorigenesis. Molecular Biology Reports 2023;50(8):6871-83.
18. 18. Hong T, Park H, An G, Song G, Lim W. Ethalfluralin induces developmental toxicity in zebrafish via oxidative stress and inflammation. Science of the Total Environment 2023;854:158780.
19. 19. Othmène YB, Monceaux K, Karoui A, Salem IB, Belhadef A, Abid-Essefi S, Lemaire C. Tebuconazole induces ROS-dependent cardiac cell toxicity by activating DNA damage and mitochondrial apoptotic pathway. Ecotoxicology and Environmental Safety 2020;204:111040.
20. 20. Weis GCC, Assmann CE, Mostardeiro VB, de Oliveira Alves A, da Rosa JR, Pillat MM, de Andrade CM, Schetinger MRC, Morsch VMM, da Cruz IBM. Chlorpyrifos pesticide promotes oxidative stress and increases inflammatory states in BV-2 microglial cells: A role in neuroinflammation. Chemosphere 2021;278:130417.
21. 21. Harry GJ. Microglia in neurodegenerative events—an initiator or a significant other? International journal of molecular sciences 2021;22(11):5818.
22. 22. Li S, Lei Z, Sun T. The role of microRNAs in neurodegenerative diseases: A review. Cell Biology and Toxicology 2023;39(1):53-83.
23. 23. Bai X, Bian Z. MicroRNA-21 is a versatile regulator and potential treatment target in central nervous system disorders. Frontiers in molecular neuroscience 2022;15:842288.
24. 24. Zhang X, Li X, Li B, Sun C, Zhang P. miR-21-5p protects hippocampal neurons of epileptic rats via inhibiting STAT3 expression. Advances in Clinical & Experimental Medicine 2020;29(7): 793-801. doi: 10.17219/acem/121929.
25. 25. Li Q, Yu P, Zeng Q, Luo B, Cai S, Hui K, Yu G, Zhu C, Chen X, Duan M. Neuroprotective effect of hydrogen-rich saline in global cerebral ischemia/reperfusion rats: up-regulated tregs and down-regulated miR-21, miR-210 and NF-κB expression. Neurochemical research 2016;41:2655-65.
26. 26. Yuan M, Yang X, Duscher D, Xiong H, Ren S, Xu X, Wang C, Chen J, Liu Y, Machens H-G. Overexpression of microRNA-21-5p prevents the oxidative stress-induced apoptosis of RSC96 cells by suppressing autophagy. Life Sciences 2020;256:118022.