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Immunomodulatory effect of imidacloprid on macrophage RAW 264.7 cells
Larissa Vivan Cestonaro1, Riciéli Pacheco Crestani2, Fernanda Mocelin Conte1
1Laboratório de Toxicologia, Departamento de Análises, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre, RS, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas (PPGCF), Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul (UFRGS), Porto Alegre RS, Brazil.
Abstract:
The neonicotinoid imidacloprid was promoted in the market because of widespread resistance to other insecticides, plus its low mammalian impact and higher specific toxicity towards insects. This study aimed to evaluate the immunomodulatory effect of imidacloprid on macrophages. RAW 264.7 cells were incubated to 0-4000 mg/L of imidacloprid for 24 and 96 h. Imidacloprid presented a concentration-dependent cytotoxicity after 24 h and 96 h incubation for MTT reduction (3-(4,5-dimethyl-thiazol-2-yl)- 2,5-diphenyltetrazolium bromide) (EC50 519.6 and 324.6 mg/L, respectively) and Neutral Red (3-amino-7-dimethylamino-2-methylphenazine hydrochloride) assays (EC50 1139.0 and 324.2 mg/L, respectively). Moreover, imidacloprid decreased the cells' inflammatory response and promoted a mitochondrial depolarization. The complex II and succinate dehydrogenase (SDH) activities in RAW 264.7 cells incubated with imidacloprid increased more at 24 h. These results suggest that imidacloprid exerts an immunomodulatory effect and mitochondria can act as regulator of innate immune responses in the cytotoxicity mediated by the insecticide in RAW 264.7 cells.
Insights
The insecticide imidacloprid shows immunomodulatory effects on macrophages, impacting their inflammatory response and mitochondrial function. This study reveals how this common pesticide affects cellular immunity and innate responses.
Area of Science:
- Immunology
- Toxicology
- Cell Biology
Background:
- Neonicotinoid insecticides like imidacloprid are widely used due to insect resistance to older chemicals.
- Imidacloprid is noted for low mammalian toxicity and high insect-specific toxicity.
- Its immunomodulatory effects on macrophages, key immune cells, require thorough investigation.
Purpose of the Study:
- To investigate the immunomodulatory impact of imidacloprid on RAW 264.7 macrophage cells.
- To assess the cytotoxicity and cellular responses induced by varying concentrations of imidacloprid.
- To explore the role of mitochondria in imidacloprid-mediated cytotoxicity.
Main Methods:
- RAW 264.7 cells were exposed to imidacloprid (0-4000 mg/L) for 24 and 96 hours.
- Cytotoxicity was evaluated using MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide) and Neutral Red assays.
- Mitochondrial depolarization and the activity of Complex II/succinate dehydrogenase (SDH) were measured.
Main Results:
- Imidacloprid exhibited concentration-dependent cytotoxicity with calculated EC50 values.
- Exposure to imidacloprid reduced the inflammatory response of macrophage cells.
- Mitochondrial depolarization was observed, and Complex II/SDH activity increased at 24 hours.
Conclusions:
- Imidacloprid exerts significant immunomodulatory effects on macrophages.
- Mitochondria play a crucial role in regulating innate immune responses during imidacloprid-induced cytotoxicity.
- These findings highlight the complex interactions between neonicotinoids and cellular immune systems.
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