Peucedanum praeruptorum Dunn leaves extract and its active coumarin praeroside Ⅴ alleviate ethanol-induced acute duodenal damage by suppressing apoptosis and ferroptosis in mice.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE: Excessive alcohol consumption can cause severe pathological damages in gastrointestinal tract, including alcoholic acute duodenal injury (ADI), with limited effective therapeutic strategy. Peucedanum praeruptorum Dunn, a traditional Chinese herb, has been used for the treatment of gastrointestinal diseases. However, the effects and active components of its leaf (PPL) on alcoholic ADI remain unclear. AIM OF THE STUDY: This study aims to investigate the mechanisms and active compounds of PPL extract against alcoholic ADI in mice. MATERIALS AND METHODS: Alcoholic ADI mouse model was established, and administered PPL extract via gavage. Apoptosis and ferroptosis were assessed using Tunel staining, Western blot, and immunohistochemistry. RNA sequencing (RNA-seq), immunofluorescence, Western blot and Elisa kits were employed to explore the underlying mechanism. The active components of PPL extract were then detected by high-performance liquid chromatography, mass spectrometry and nuclear magnetic resonance. RESULTS: PPL extract effectively mitigated alcohol-induced acute duodenal injuries in mice. Importantly, PPL extract administration reduced alcohol-induced cell apoptosis and ferroptosis in duodenal tissues in mice. RNA-seq analysis demonstrated a strong correlation with IL-17 pathway after PPL pretreatment in mice with ADI. Western blots results showed that PPL extract administration inhibited alcohol-induced up-regulation of the related proteins of IL-17/MAPKs signaling pathways. Notably, HPLC, MS and NMR analysis identified praeroside Ⅴ as the main component of PPL, and praeroside Ⅴ relieved ethanol-induced ADI in mice. In addition, molecular docking and dynamics simulation revealed that praeroside Ⅴ showed stable affinity binding to IL-17A, TNF-a and GPX4. Western blots analysis showed that praeroside Ⅴ reduced the expressions of IL-17A and TNF-a, and increased the level of GPX4 in ethanol-treated mice. CONCLUSIONS: Pretreatment with PPL extract alleviated alcoholic ADI by inhibiting cell apoptosis and ferroptosis, possibly through suppression of IL-17/MAPK pathways, and praeroside Ⅴ was the main active component of PPL extract. These data supported the development and utilization of PPL and praeroside Ⅴ in gastrointestinal prevention and treatment.