Olaparib Blocks Oxidative Stress-Dependent NLRP3 Inflammasome in Pediatric Acute Lung Injury by Metabolic Reprogramming via AMPK-Driven Autophagic Flux.

In vitro and in vivo experimental study using macrophage cell lines and juvenile mice

“This study aims to evaluate the therapeutic efficacy and mechanistic basis of Olaparib, a clinically approved PARP inhibitor, using in vitro and in vivo models of pediatric ALI.”

LPS/ATP-stimulated THP-1-derived macrophages and 3-4-week-old male C57BL/6J mice

“This study aims to evaluate the therapeutic potential of Olaparib in LPS‐induced 3–4‐week‐old C57BL/6J mice and LPS/ATP‐stimulated THP‐1‐derived macrophages”

Six mice per experimental group

“After a 1‐week acclimatization period, mice were randomly allocated into five experimental groups (n = 6 per group)”

24 hours post-LPS administration for in vivo analysis

“At 24 h post‐LPS administration, mice were euthanized via cervical dislocation under pentobarbital sodium anesthesia (100 mg/kg, i.p.).”

Olaparib induced metabolic reprogramming and activated the AMPK/ULK1 signaling pathway, enhancing autophagic flux in macrophages

“In LPS/ATP‐stimulated THP‐1‐derived macrophages, Olaparib induced metabolic reprogramming, as indicated by a reduced NAD+/NADH ratio and an elevated AMP/ATP ratio, leading to activation of the AMPK/ULK1 signaling pathway.”

Olaparib reduced oxidative stress and suppressed NLRP3 inflammasome activation in both cell and mouse models

“Enhanced autophagy was associated with attenuation of oxidative stress, including reductions in mitochondrial and intracellular reactive oxygen species and malondialdehyde levels, and suppression of NLRP3 inflammasome activation, as evidenced by decreased cleaved caspase‐1, IL‐1β, and IL‐18.”

Olaparib administration reduced lung injury, inflammation, and apoptosis while restoring alveolar-capillary barrier integrity in mice

“In a juvenile murine model of LPS‐induced ALI, Olaparib administration reduced histopathological lung injury, inflammatory cell infiltration, and pulmonary edema.”

Use of compound C as an AMPK inhibitor has known off-target effects, and species-specific differences may limit extrapolation to humans

“A major limitation is the use of compound C as a pharmacological AMPK inhibitor, which is associated with known off‐target effects on multiple kinases.”