Peptidomics Landscape of Intrahepatic Cholestasis of Pregnancy and its Molecular Mechanism Investigation

Intrahepatic cholestasis of pregnancy (ICP) is a liver disorder that poses substantial health risks to both mother and fetus. However, its pathogenesis remains unclear. Our previous study highlighted the significance of ferroptosis in ICP. To fully elucidate the pathogenesis of ferroptosis related to ICP and identify new therapeutic strategies, we used nanoscale liquid chromatography coupled with tandem mass spectrometry analysis in an in vitro model of ICP induced by taurocholic acid to investigate the peptidomics of ICP. The peptide profile successfully revealed 27,527 peptides, of which 1,180 were related to ferroptotic differentially expressed peptides in the screened ICP. Through comprehensive bioinformatics analyses, combined with 3,783 ICP and 1,518 ferroptosis targets, we identified 20 hub genes and molecular functions, including ubiquitin protein ligase binding, which may be involved in the development of ICP. Four potential bioactive peptides were selected through in silico analysis and further verified using in vitro, molecular docking, and molecular dynamics studies. Peptides DLTGIPPAPR and PRLPEEWSQW have the potential to enhance trophoblast viability and modulate important genes, such as HSPB1, HSPA5, and SQSTM1, which could be beneficial in managing ICP by inhibiting ferroptosis. Our findings provide crucial insight into the pathophysiological mechanisms underlying ICP.