Cypermethrin induces apoptosis, autophagy and inflammation via ERS-ROS-NF-κB axis in hepatocytes of carp (Cyprinus carpio)

Cypermethrin (CYP, IUPAC name: [cyano-(3-phenoxyphenyl)methyl] 3-(2,2-dichloroethenyl)-2,2-dimethylcy-clopropane-1-carboxylate) is a pyrethroid insecticide that poses a threat to the health of humans and aquatic animals due to its widespread use and environmental contamination. However, the mechanism of CYP on apoptosis, autophagy and inflammation in hepatocytes of carp (Cyprinus carpio) is unknown. We hypothesized that CYP caused damage to hepatocytes through the endoplasmic reticulum stress (ERS) pathway, CCK-8 was used to detect the toxic effects of different doses of CYP on hepatocytes, and finally low (L, 10 mu M), medium (M, 40 mu M), and high (H, 80 mu M) doses of CYP was selected to construct the model. ROS staining, oxidative stress-related indices (MDA, CAT, T-AOC, SOD), AO/EB staining, MDC staining, and the expression levels of related genes were detected using qRT-PCR and western blot. Our results showed that CYP exposure resulted in an increase in ROS production, an increase in MDA content, and a decrease in the activity of CAT, SOD, and T-AOC in hepatocytes; the proportion of apoptotic, necrotic, and autophagic cells increased significantly in a dose-dependent manner. We also found that CYP exposure increased the expression levels of endoplasmic reticulum-related genes (GRP78, PERK, IRE-1, ATF-6 and CHOP), apoptosis (Bcl-2, Bax, Caspase-3, Caspase-9 and Cyt-c) and autophagy-related genes (LC3b, Beclin1 and P62) also showed dose-dependent changes, and the expression levels of inflammation-related genes (NF-kappa B, TNF-alpha, IL-1 beta, IL-6) were also significantly elevated. Thus, we demonstrated that CYP exposure caused apoptosis, autophagy and inflammation in hepatocytes via ERS-ROS-NF-kappa B axis. This research contributes to our understanding of the molecular mechanisms underlying CYP-induced damage in hepatocytes of carp (Cyprinus carpio).

Automated summary

This study found that CYP exposure caused endoplasmic reticulum stress in carp hepatocytes, resulting in apoptosis, autophagy, and inflammation. The expression levels of related genes also showed dose-dependent changes.