2. Academic Validation
  3. Naoxintong capsule accelerates mitophagy in cerebral ischemia-reperfusion injury via TP53/PINK1/PRKN pathway based on network pharmacology analysis and experimental validation

Naoxintong capsule accelerates mitophagy in cerebral ischemia-reperfusion injury via TP53/PINK1/PRKN pathway based on network pharmacology analysis and experimental validation

  • J Ethnopharmacol. 2025 Jan 10:336:118721. doi: 10.1016/j.jep.2024.118721.
Jinfeng Shang 1,  Yinlian Wen 2,  Xiaolu Zhang 3,  Guijinfeng Huang 4,  Wenbin Chen 5,  Bohong Wang 6,  Kai Wu 7,  Quan Xiang 8,  Xin Liu 9
Affiliations

Affiliations

  • 1 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: sjf01qtdd@163.com.
  • 2 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: wenyinlian99@163.com.
  • 3 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: swagdeer@163.com.
  • 4 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: hgjf2000@163.com.
  • 5 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: wenbin990915@126.com.
  • 6 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: 17609078266@163.com.
  • 7 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: wukaiLX@163.com.
  • 8 Gansu University of Chinese Medicine, Gansu 730101, China. Electronic address: quanxiang521@163.com.
  • 9 Beijing University of Chinese Medicine, Beijing 102488, China. Electronic address: liuxin1011@bucm.edu.cn.
PMID: 39173723 DOI: 10.1016/j.jep.2024.118721
Abstract

Ethnopharmacological relevance: The incidence and mortality of cerebrovascular diseases are increasing year by year. Cerebral ischemia-reperfusion injury (CIRI) is common in patients with ischemic stroke. Naoxintong (NXT) is composed of a variety of Chinese medicines and has the ability to treat CIRI.

Aim of the study: The aim of this study is to investigate whether NXT regulates Mitophagy in CIRI based on network pharmacology analysis and experimental validation.

Materials and methods: Oxygen and glucose deprivation/re-oxygenation (OGD/R, 2/22 h) model of PC12 cells and transient middle cerebral artery occlusion (tMCAO, 2/22 h) model of rats were established. Pharmacodynamic Indicators include neurological deficit score, 2,3,5-triphenyte-trazoliumchloride (TTC) staining, hematoxylin-eosin (HE) staining and cell viability. Network pharmacology was used to predict pharmacological mechanisms. Pharmacological mechanism indexes include transmission electron microscopy (TEM), drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), immunohistochemistry (IHC), western blot (WB) and immunofluorescence (IF). Kevetrin (an agonists of p53) and pifithrin-α (an inhibitor of p53) used to detect the key role of p53 in Mitophagy of NXT.

Results: NXT (1% serum containing NXT and 110 mg/kg) improved the damage of OGD/R PC12 cells and tMCAO rats, and this protective effect was related to the anti-oxidation and ability to promote Mitophagy of NXT. NXT and pifithrin-α increased the expression of promoting-mitophagy targets (PINK1, PRKN and LC3B) and inhibited the expression of inhibiting-mitophagy targets (p52) via restraining p53, and finally accelerated Mitophagy caused by CIRI.

Conclusion: This study demonstrates that NXT promotes Mitophagy in CIRI through restraining p53 and promoting PINK1/PRKN in vivo and in vitro.

Keywords

Cerebral ischemia-reperfusion injury; Mitophagy; Naoxintong capsule; TP53/PINK1/PRKN pathway.

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