The presence of acute pancreatitis-induced pancreatic necrosis and systemic inflammation is closely related to the prognosis and severity of the disease. Pentraxin 3 is a mediator that aggravates cell injury, and its activation is closely related to oxidative stress and reactive oxygen species generation. This study aims to explore whether PTX3 regulates AP pancreatic necrosis and reveal its potential mechanism of action. By using AP time gradient transcriptomics, proteomics, and liquid chip combined analysis methods, we screened and found a close association between PTX3 and AP. Subsequently, we constructed a classical cerulein-induced in vivo AP model and utilized a CCK-induced in vitro acinar cell injury model to determine the expression of PTX3 and its regulatory role on AP. The results showed that PTX3 presented a high expression level in the cerulein-induced AP model; while after the intervention of recombinant protein PTX3, more severe pancreatic tissue damage and increased serum amylase were observed; in addition, the expression level of PTX3 was significantly higher than that of other proteins, mitochondrial membrane potential decreased and reactive oxygen species levels increased in the in vitro model and were regulated through the oxidative phosphorylation pathway. In conclusion, multi-omics integrated analysis confirmed the close association between Pentraxin 3 and AP and revealed that it exacerbates acute pancreatitis acinar cell injury by mediating oxidative phosphorylation processes.