Mixed plasticizers aggravated apoptosis by NOD2-RIP2-NF-κB pathway in grass carp hepatocytes

The wide application of plastics led to the wide exposure of plasticizers to the environment. As a new environmental pollutant, plasticizers' toxicity researches were far from enough in fish. To further explore these mechanisms, we used two common plasticizers (Diethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP) expose to grass carp hepatocytes (L8824). The results showed that the mRNA levels of NOD2-RIP2-NF-κB signal pathway and its downstream inflammatory genes were significantly increased compared to those in control group. Then, the levels of mRNAs and proteins of apoptosis markers were changed, and hepatocytes apoptosis was induced. After DBP and DEHP exposure together, there were higher levels of inflammatory factors and the proportion of apoptotic cells. After NOD2 inhibitor treatment, the phenomena mentioned above were obviously alleviated. We conclude that DBP and DEHP exposure at least partially activated the NOD2-RIP2-NF-κB signal pathway in grass carp hepatocytes, and caused inflammation and apoptosis. In terms of hepatotoxicity, there was synergistic relationship between DBP and DEHP. In addition, we put forward new views on the use of plasticizers: select low toxicity plasticizers, then reduce the types of plasticizers used and reduce the high toxicity level of mixed plasticizers.