Costunolide: Targeting endothelial cell PANoptosis to mitigate lung injury in acute pancreatitis mice

Background

Acute pancreatitis (AP) presents several critical risks, among which acute lung injury (ALI) is one of the most severe complications. Previous research has highlighted the anti-inflammatory properties of costunolide (COS); however, its potential to prevent AP-associated ALI remains unexplored. Here, we hypothesize that COS can mitigate AP-induced lung injury by reducing endothelial cell PANoptosis-a form of programmed cell death.

Purpose

The work aimed to examine the therapeutic effect and mechanism of COS on lung injury in AP mice, with particular emphasis on its impact on endothelial cell PANoptosis.

Study design

The AP mouse model was created with partial pancreatic duct ligation followed by a single injection of caerulein, and the therapeutic benefits of varying dosages of COS on these AP mice were evaluated. Dexamethasone served as the positive control. The endothelial cell damage caused by CIRP served as an in vitro model to assess the impact of varying doses of COS on PANoptosis.

Methods

Initially, the toxicological impact of varying doses of COS on the main organs of healthy mice was assessed, subsequently examining the inhibitory effects of COS on pancreatic and pulmonary damage, inflammatory response, and lung tissue PANoptosis in AP mice. Transcriptomic sequencing of pancreatic and pulmonary tissue was used to identify critical targets for COS intervention in AP. To investigate if CIRP activation can elicit PANoptosis and the inhibitory effects of COS in human umbilical vein endothelial cells, the regulatory impact of COS on ALDH2 was assessed via gene knockdown, western blot, and ELISA.

Results

COS demonstrated substantial anti-inflammatory and lung-protective effects in AP mice. Notably, COS significantly downregulated the expression of PANoptosis markers in the lung tissue of AP mice. Cold-induced RNA-binding proteins (a damage-associated molecular pattern that increases during AP) cause increased production of PANoptosis markers in human umbilical vein endothelial cells, while COS exerted a dose-dependent inhibitory effect on these markers. Single-cell RNA sequencing and bulk transcriptomic analysis, Mendelian randomization, molecular docking and molecular dynamics simulation identified alcohol dehydrogenase 2 (ALDH2) as a possible target for COS intervention in AP-related ALI. Subsequent lentiviral transfection, ELISA,‌ and western blot assays confirmed that COS's PANoptosis-inhibitory effect in endothelial cells is partially dependent on ALDH2 activity and expression.

Conclusion

Our findings indicate that COS, a novel ALDH2 activator, may treat AP-associated ALI by inhibiting ZBP1-mediated PANoptosis in endothelial cells, thereby establishing a theoretical basis for the clinical use of COS and identifying an entirely novel target for AP-associated ALI treatment.