Multi-omics study reveals Shuangshen Pingfei formula regulates EETs metabolic reprogramming to exert its therapeutic effect on pulmonary fibrosis.

As a clinical formula derived from Renshen Pingfei San, Shuangshen Pingfei formula (SSPF) has been used to treat pulmonary fibrosis (PF). However, its in-depth mechanism of action remains unknown. In this study, the effect of SSPF was evaluated by applying a rat model of PF caused by intratracheal drip bleomycin. To characterize the molecular changes related to PF and reveal therapeutic targets for SSPF, we performed transcriptomic and metabolomic analyses on rat lung. Finally, western blotting and qPCR experiments were used to validate the multi-omics results. As a result, a significant reduction in inflammation and fibrosis caused by BLM was observed when SSPF was administered. Widespread changes in gene expression and metabolic programming were observed in the lungs of PF rats through RNA-seq and untargeted metabolomic analysis. Combined transcriptomic and metabolomic analyses revealed the involvement of arachidonic acid (AA) metabolism pathways in PF. Further validation of AA metabolite synthase genes and protein levels showed a significant decrease in the levels of epoxyeicosatrienoic acids (EETs) synthases, including Cyp2j2, Cyp2b1, in the PF lungs. SSPF treatment regulated the above changes in gene expression and metabolic programming, particularly the regulation of EETs. This study is the first to investigate the mechanism of action of SSPF in the treatment of PF from the perspective of regulating the synthesis of EETs in AA metabolism.