The toxic effects of imidacloprid exposure on HepG2 cell based on non-targeted metabolomics

Xingfan Zhou; Yiran Sun; Xiaojun Zhu; Mengwen Lin; Wenlin Bai; Yingying Zhang; Wenping Zhang

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The toxic effects of imidacloprid exposure on HepG2 cell based on non-targeted metabolomics

VernacularTitle:基于非靶向代谢组学的吡虫啉暴露致人肝细胞HepG2毒性效应研究
Author: Xingfan ZHOU ¹ ; Yiran SUN ² ; Xiaojun ZHU ³ ; Mengwen LIN ² ; Wenlin BAI ² ; Yingying ZHANG ² ; Wenping ZHANG ²
Author Information

1. Beijing Key Laboratory of Occupational Safety and Health, Institute of Urban Safety and Environmental Science, Beijing Academy of Science and Technology, Beijing 100054, China.
2. School of Public Health, Shanxi Medical University, Taiyuan, Shanxi 030001, China.
3. National Center for Occupational Safety and Health, NHC, Beijing 102308, China.
Publication Type:Experiment
Keywords: imidacloprid; HepG2 cell; non-targeted metabolomics; mTOR signaling pathway; galactose metabolism
From: Journal of Environmental and Occupational Medicine 2023;40(2):216-223
CountryChina
Language:Chinese
Abstract: Background Imidacloprid is a neonicotinoid insecticide that is widely used in agricultural production, with a high detection rate in human biological samples. Previous studies have shown a high correlation between imidacloprid exposure and liver injury, but the specific mechanism is still unknown. Objective To observe potential toxic effects of HepG2 cells and its perturbation of non-targeted metabolic profile after imidacloprid exposure, and to explore possible molecular mechanisms of hepatotoxicity of imidacloprid by analyzing invovlved biological processes and signaling pathways. Methods HepG2 cell suspension was prepared and seeded in a 96-well plate, which was divided into blank control group, dimethyl sulfoxide (DMSO) solvent control group and imidacloprid exposure groups with multiple concentrations. Each group was set with 5 parallel samples. The viability of HepG2 cells viability were determined after 8 h of exposure to different concentrationsof imidacloprid (1, 2.5, 5, 7.5, 10 mmol·L⁻¹), and the dose-effect relationship was analyzed. A proper concentration (3 mmol·L⁻¹ with 80% viability) was chosen for imidacloprid exposure, non-targeted metabolomic analysis was applied to the cultivated HepG2 cells using UHPLC-Q-TOF/MS technology, the differential metabolites between groups were screened, and the bioprocess and related signaling pathways of their enrichment were annotated using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results Compared to the other two groups, the survival rates of HepG2 cells in the imidacloprid exposure groups decreased. A survival rate of about 86% of HepG2 cells was found in HepG2 cells exposed to 2.5 mmol·L⁻¹ imidacloprid exposure. The non-targeted metabolomics studies showed that 61 metabolites were significantly affected in HepG2 cells after 3 mmol·L⁻¹ imidacloprid exposure, including creatine (variable importance in projection VIP=1.11, P<0.001), arginine (VIP=1.47, P=0.048), taurine (VIP=4.28, P=0.001), and α-D-glucose (VIP=1.90, P=0.006). The differential metabolites enriched in bioprocess and related signaling pathways were mainly directed to mTOR signaling pathways (P<0.001), arginine and proline metabolism (P=0.002), and galactose metabolism (P=0.015). Conclusion Imidacloprid exposure can significantly inhibit the survival rate of HepG2 cells, and interfere with the mTOR signaling pathway, arginine and proline metabolism, galactose metabolism, and so on.