2. Academic Validation
  3. Obacunone inhibits RIPK1/RIPK3/MLKL-mediated necroptosis by suppressing mitochondrial ROS and MIF signaling in intestinal ischemia-reperfusion injury

Obacunone inhibits RIPK1/RIPK3/MLKL-mediated necroptosis by suppressing mitochondrial ROS and MIF signaling in intestinal ischemia-reperfusion injury

  • Int Immunopharmacol. 2026 Jan 1;168(Pt 2):115901. doi: 10.1016/j.intimp.2025.115901.
Xiao-Yao Li 1,  Jing-Yuan Xie 1,  Yu-Tong Yin 1,  Xiao-Guo Weng 1,  Zi-Yi Huang 1,  Lu-Si Wang 1,  Ke Ding 1,  Rong Chen 2,  Qing-Tao Meng 3
Affiliations

Affiliations

  • 1 Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, China.
  • 2 Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, China; Department of Anesthesiology, East Hospital, Renmin Hospital of Wuhan University, Wuhan, China.
  • 3 Department of Anesthesiology, Renmin Hospital of Wuhan University, Wuhan, China; Department of Anesthesiology, East Hospital, Renmin Hospital of Wuhan University, Wuhan, China. Electronic address: [email protected].
PMID: 41317539 DOI: 10.1016/j.intimp.2025.115901
Abstract

Background: Intestinal ischemia-reperfusion (IIR) is a critical pathophysiological event associated with high morbidity and mortality, often leading to intestinal barrier disruption and systemic inflammatory responses. Obacunone (OBA), a naturally occurring limonoid compound with documented antioxidant and anti-inflammatory properties, has not been previously studied in the context of IIR. This study aimed to evaluate its protective effects against IIR-induced damage.

Methods: In vivo IIR mouse models and in vitro IEC-6 cell models subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) were used to assess the protective effects of OBA. Histological analysis, immunohistochemistry, immunofluorescence and western blotting were performed to evaluate tissue injury and Necroptosis. Mitochondrial function and ROS production were assessed by JC-1 staining, MitoSOX fluorescence, and transmission electron microscopy. Network pharmacology and molecular docking were employed to identify potential OBA targets, followed by pharmacological validation using the MIF inhibitor ISO-1 and recombinant MIF protein.

Results: OBA pre-treatment significantly alleviated IIR-induced intestinal damage, restored epithelial integrity, and suppressed the phosphorylation of RIPK1, RIPK3, and MLKL. OBA attenuated ROS accumulation, suppressed mitochondrial ROS (mtROS) production, and preserved mitochondrial morphology and membrane potential, thereby suppressing Necroptosis. Network pharmacology and molecular docking identified macrophage migration inhibitory factor (MIF) as a potential target of OBA. MIF inhibition mimicked OBA's protective effects, whereas exogenous MIF partially reversed them, indicating that OBA suppresses Necroptosis via dual regulation of mtROS and MIF signaling.

Conclusion: OBA protects against IIR-induced epithelial injury partly through modulation of mtROS- and MIF-driven Necroptosis, providing a promising therapeutic strategy for intestinal ischemia-reperfusion injury and related inflammatory disorders.

Keywords

Intestinal ischemia-reperfusion injury; MIF; Necroptosis; Network pharmacology; Obacunone; mtROS.

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