Punicalagin inhibits excessive autophagy and improves cerebral function in neonatal rats with hypoxia-ischemia brain injury by regulating AKT-FOXO4

Ming Shen; Junhong Lu; Caiyan Li; Yujiang Li; Qianqian Yu; Xinyu Gao; Zhouguang Wang; Guanhu Yang; Shengcun Li; Zhenlang Lin

doi:10.1016/j.phymed.2024.156330

Punicalagin inhibits excessive autophagy and improves cerebral function in neonatal rats with hypoxia-ischemia brain injury by regulating AKT-FOXO4

Phytomedicine. 2025 Jan 1:136:156330. doi: 10.1016/j.phymed.2024.156330. Online ahead of print.

Authors

Ming Shen¹, Junhong Lu², Caiyan Li², Yujiang Li², Qianqian Yu³, Xinyu Gao², Zhouguang Wang², Guanhu Yang⁴, Shengcun Li⁵, Zhenlang Lin⁶

Affiliations

¹ Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, Zhejiang, PR China; Key Laboratory of Rehabilitation, Key Laboratory of Perinatal Medicine of Wenzhou, Wenzhou 325024, Zhejiang, China.
² Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, Zhejiang, PR China.
³ Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Key Laboratory of Rehabilitation, Key Laboratory of Perinatal Medicine of Wenzhou, Wenzhou 325024, Zhejiang, China.
⁴ Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Department of Specialty Medicine, Ohio University, 45701,Athens, OH, United States.
⁵ Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou 325000, Zhejiang, PR China; Department of Specialty Medicine, Ohio University, 45701,Athens, OH, United States. Electronic address: [email protected].
⁶ Department of Neonatology, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, Zhejiang, China; Key Laboratory of Rehabilitation, Key Laboratory of Perinatal Medicine of Wenzhou, Wenzhou 325024, Zhejiang, China. Electronic address: [email protected].

PMID: 39756314
DOI: 10.1016/j.phymed.2024.156330

Abstract

Background: Neonatal hypoxic-ischemic encephalopathy (HIE) has a high incidence and mortality rate, representing a significant patient burden. Therefore, treatment strategies that work synergistically with hypothermic therapies are urgently required. Punicalagin (PUN) is a natural and safe polyphenol with anti-inflammatory functions whose excellent water solubility and safety make it an advantageous perinatal medication. However, its underlying mechanisms of action in HIE remain unclear.

Objectives: This study investigated the role and associated mechanism of action PUN in HIE.

Methods: We used the Rice Vannucci method to construct an in vivo HIE model in rats, from which we extracted primary cortical neurons to construct an in vitro oxygen and glucose deprivation/reoxygenation (OGD/R) model. The mechanisms of action of PUN were investigated using transcriptome sequencing, laser speckle contrast imaging, 2,3,5-triphenyltetrazolium chloride-staining, the Morris water maze test, western blotting, qPCR, immunofluorescence, and histochemistry.

Results: HIE rats demonstrated excessive autophagy and inflammation. PUN reduced brain tissue damage and neuronal apoptosis, and improved cerebral blood flow perfusion, learning, and cognitive abilities. PUN attenuated autophagic overexpression following HIE and inhibited the AKT-FOXO4 (forkhead box O4) signaling pathway. The neuroprotective effects of PUN were inhibited by treatment with the AKT signaling pathway and autophagy inhibitor 3-MA. Furthermore, brain tissue damage was significant and PUN was ineffective in siFOXO4 rats.

Conclusions: PUN significantly reduces cerebral infarction, neuroinflammation, and excessive autophagy caused by HIE, thereby exerting short- and long-term neuroprotective effects. Mechanistically, the neuroprotective effect of PUN is mediated by activation of the AKT-FOXO4 pathway. Therefore, PUN may be a potential therapy for HIE.

Keywords: Autophagy; Inflammation; Neonatal hypoxic-ischemic encephalopathy; Punicalagin.