Tannic acid through ROS/TNF-α/TNFR 1 antagonizes atrazine induced apoptosis, programmed necrosis and immune dysfunction of grass carp hepatocytes

Atrazine (ATR) is a commonly used triazine herbicide, which will remain in the water source, soil and biological muscle tissue for a long time, threatening the survival of related organisms and future generations. Tannic acid (TAN), a glucosyl compound found in gallnuts, has previously been shown to antagonize heavy metal toxicity, antioxidant activity, and inflammation. However, it is unclear whether TAN can antagonize ATR-induced Grass carp hepatocytes (L8824 cells) cytotoxicity. Therefore, we treated L8824 cells with 3 mu g mL(-1) ATR for 24 h to establish a toxic group model. The experimental data of flow cytometry and AO/EB staining together showed that the ratio of apoptosis and necrosis in L8824 cells after ATR exposure was significantly higher than that in the control group. Furthermore, RT-qPCR showed that inflammatory factors (TNF-alpha, IL-1 beta, IL-6, INF-gamma) were upregulated and antimicrobial peptides (hepcidin, beta-defensin and LEAP2) were induced down-regulated in L8824 cells, leading to immune dysfunction. The measurement results of oxidative stress-related indicators showed that the levels of ROS and MDA increased after ATR exposure, the overall anti-oxidative system was down-regulated. Western blotting confirmed that TNF-alpha/TNFR 1-related genes were also up-regulated. This indicates that ATR stimulates oxidative stress in L8824 cells, which in turn promotes the binding of TNF-alpha to TNFR 1. In addition, TRADD, FADD, Caspase-3, P53, RIP1, RIP3 and MLKL were found to be significantly upregulated by Western blotting and RT-qPCR. Conditioned after ATR exposure compared to controls. It indicates that ATR activates apoptosis and necrosis of TNF-alpha/TNFR 1 pathway by inducing oxidative stress in L8824 cells. Furthermore, the use of TAN (5 mu M) significantly alleviated the toxic effects of ATR on L8824 cells mentioned above. In conclusion, TAN restrains ATR-induced apoptosis, programmed necrosis and immune dysfunction through the ROS/TNF-alpha/TNFR 1 pathway.

Automated summary

Atrazine, a commonly used herbicide, poses a threat to organisms and future generations by remaining in water sources, soil, and biological tissue. Tannic acid, found in gallnuts, has been shown to counteract heavy metal toxicity, antioxidant activity, and inflammation. This study investigates whether tannic acid can alleviate Atrazine-induced cytotoxicity and immune dysfunction in Grass carp hepatocytes. The results show that Atrazine exposure induces apoptosis, inflammation, and oxidative stress in the cells, but tannic acid treatment alleviates the toxic effects through the ROS/TNF-alpha/TNFR 1 pathway.