Unravelling the Polytoxicology of Chlorfenapyr on Non-Target HepG2 Cells: The Involvement of Mitochondria-Mediated Programmed Cell Death and DNA Damage

Abstract

Chlorfenapyr (CHL) is a type of insecticide with a wide range of insecticidal activities and unique targets. The extensive use of pesticides has caused an increase in potential risks to the environment and human health. However, the potential toxicity of CHL and its mechanisms of action on humans remain unclear. Therefore, human liver cells (HepG2) were used to investigate the cytotoxic effect and mechanism of toxicity of CHL at the cellular level. The results showed that CHL induced cellular toxicity in HepG2 cells and induced mitochondrial damage associated with reactive oxygen species (ROS) accumulation and mitochondrial calcium overload, ultimately leading to apoptosis and autophagy in HepG2 cells. Typical apoptotic changes occurred, including a decline in the mitochondrial membrane potential, the promotion of Bax/Bcl-2 expression causing the release of cyt-c into the cytosol, the activation of cas-9/-3, and the cleavage of PARP. The autophagic effects included the formation of autophagic vacuoles, accumulation of Beclin-1, transformation of LC3-II, and downregulation of p62. Additionally, DNA damage and cell cycle arrest were detected in CHL-treated cells. These results show that CHL induced cytotoxicity associated with mitochondria-mediated programmed cell death (PCD) and DNA damage in HepG2 cells.

Mitochondria-Mediated Apoptosis and Autophagy Participate in Buprofezin-Induced Toxic Effects in Non-Target A549 Cells

“…The fixed cells were dried using graded acetone and embedded in Epon 812 (Fluka, Buchs, Switzerland). Ultrathin sections were dyed with lead citrate and uranyl acetate (Macklin, Shanghai, China), then observed using a transmission electron microscope [ 22 , 23 ]. The constitution of the autophagosomes was detected by monodansylcadaverine (MDC) (Beyotime, Shanghai, China) dyes.…”

“…The constitution of the autophagosomes was detected by monodansylcadaverine (MDC) (Beyotime, Shanghai, China) dyes. Cells were hatched with 1 μg/mL MDC in the dark for 30 min and then washed twice with PBS before watched by fluorescence microscope (Leica, Wetzlar, Germany) [ 23 ].…”

“…The 0.1% DMSO-treated cells were used as a control. Then, the DCFH-DA (Keygen, Nanjing, China) was used to analyse intracellular ROS generation [ 23 ]. Cells were harvested by centrifuge (2000× g , 3 min) and washed two times with PBS, then hatched with 10 μM DCFH-DA for 30 min at 37 °C.…”

“…Then, the stained cells were washed with Hanks’ buffer (Beyotime, Shanghai, China) and further hatched with PBS for 30 min at 37 °C before being detected with a flow cytometer (BD CantoⅡ, Franklin Lakes, NJ, USA). The increased Ca 2+ level was statistically analysed using the green and red fluorescence intensities by FlowJo (V10, 2015) software (BD, Ashland, OR, USA) [ 23 ].…”

Residual Behavior and Dietary Risk Assessment of Chlorfenapyr and Its Metabolites in Radish

“…The irrational use of pesticides caused by the resistance of insect pests [ 7 , 8 , 9 ] leads to the increasingly serious residual contamination of chlorfenapyr [ 10 , 11 , 12 , 13 ] in vegetables and other agricultural products. This poses a potential threat to non-target organisms [ 14 , 15 ] and human health [ 16 , 17 ]. In addition, chlorfenapyr is also degraded and metabolized in plants to produce a product of toxicological significance, tralopyril.…”