Albicanol modulates oxidative stress and the p53 axis to suppress profenofos induced genotoxicity in grass carp hepatocytes.

The organophosphorus pesticide profenofos (PFF) is widely used as an environmental contaminant, and it can remain in water bodies causing serious harm to aquatic organisms. Albicanol is a sesquiterpenoid with potent antioxidant and antagonistic activities against heavy metal toxicity. However, the mechanism of PFF induced genotoxicity in fish hepatocytes and the role Albicanol can play in this process are unknown. In this study, the model was established by treating grass carp hepatocytes with PFF (150 µM) and/or Albicanol (5 × 10-5 µg mL-1) for 24 h. The results showed that PFF exposure arrested L8824 cells in the G1-S phase. PFF caused the increase of MDA level in L8824 cells, while the decrease of SOD, CAT and T-AOC levels caused oxidative stress. Elevated levels of γH2AX, tail moment, tail length, % DNA and 8-OHdG indicated that PFF caused DNA damage in L8824 cells. PFF inhibited the expression levels of cell cycle related regulatory genes (cyclin A, cyclin D, cyclin E, CDK2 and CDK4) by upregulating p53/p21 genes and activating the p53 signaling pathway. Albicanol was used to significantly reduce the above effects caused by PFF exposure on hepatocytes in grass carp. Albicanol could reduce the increase in the proportion of cells in the G1-S phase caused by PFF. In summary, Albicanol could inhibit the genotoxicity of L8824 cells resulted from PFF exposure by decreasing oxidative stress and the p53 pathway.

Albicanol inhibits the toxicity of profenofos to grass carp hepatocytes cells through the ROS/PTEN/PI3K/AKT axis.

Profenofos (PFF) as an environmental pollutant seriously harms the health of aquatic animals, and even endangers human safety through the food chain. Albicanol, a sesquiterpenoid extraction from the Dryopteris fragrans, has previously been shown to effectively exhibit anti-aging, anti-oxidant, and antagonize the toxicity of heavy metals. However, the mechanism of hepatocyte toxicity caused by PFF and the role that Albicanol plays in this process are still unclear. In this study, a PFF poisoning model was established by treating grass carp hepatocytes cells with PFF (150 µM) for 24 h The results of AO/EB staining, Tunel staining and flow cytometry showed that the proportion of apoptotic liver cells increased significantly after exposure. The results of ROS staining show that compared with the control group, ROS levels and PTEN/PI3K/AKT-related gene expression were up-regulated after PFF exposure. RT-qPCR and Western blotting results showed that the expression of PTEN/PI3K/AKT related genes was up-regulated. These results indicate that PFF can induce oxidative stress in hepatocytes and inhibit the phosphorylation of AKT. We further found that the expressions of Bax, CytC, Caspase-3, Caspase-9, Caspase-8 and TNFR1 after PFF exposure were significantly higher than those of the control group, and Bcl-2/Bax was significantly lower than that of the control group. These results indicate that PFF can induce oxidative stress in hepatocytes and inhibit the phosphorylation of AKT and activate mitochondrial apoptosis. Using Albicanol (5 × 10-5 µg mL-1) can significantly reduce the above-mentioned effects of PFF exposure on grass carp hepatocytes cells. In summary, Albicanol inhibits PFF-induced apoptosis by regulating the ROS/PTEN/PI3K/AKT pathway.