The integration strategy of DiDang Decoction against cerebral ischemia-reperfusion injury: Regulation of apoptosis-autophagy-inflammation network and validation of PI3K/Akt pathway

Zhi-Guang Song; Yan-Song Liu; Si-Si Dai; Shu-Ting Yin; Yi-di Zeng; Yu-Chen Liu; Ji-Yong Liu; Jin-Xia Li; Yu-Yan Long; Yu-Jing Feng; Hao-Bo Jiang; Kun Lian; Kai-Ni Mao; Cai-Xing Zheng; Li-Hua Lin; Zhe-Yuan Zhang; Wen-Lu Yu; Hua Li; Wang-Hua Liu

doi:10.1016/j.jep.2025.120277

The integration strategy of DiDang Decoction against cerebral ischemia-reperfusion injury: Regulation of apoptosis-autophagy-inflammation network and validation of PI3K/Akt pathway

J Ethnopharmacol. 2025 Jul 9:120277. doi: 10.1016/j.jep.2025.120277. Online ahead of print.

Authors

Zhi-Guang Song¹, Yan-Song Liu², Si-Si Dai², Shu-Ting Yin², Yi-di Zeng³, Yu-Chen Liu², Ji-Yong Liu², Jin-Xia Li², Yu-Yan Long², Yu-Jing Feng³, Hao-Bo Jiang², Kun Lian², Kai-Ni Mao³, Cai-Xing Zheng⁴, Li-Hua Lin², Zhe-Yuan Zhang², Wen-Lu Yu², Hua Li⁵, Wang-Hua Liu⁶

Affiliations

¹ Key Laboratory of TCM Diagnostics of Hunan Provine, Changsha 410208, Hunan, China; Hunan Engineering Technology Research Centerfor Medicinal and Functional Food, Changsha,410208, Hunan, China; Key Laboratory of TCM Heart and Lung SyndromeDifferentiation & Medicated Diet and Dietotherapy,Changsha 410208, Hunan, China.
² Key Laboratory of TCM Diagnostics of Hunan Provine, Changsha 410208, Hunan, China.
³ Hunan Engineering Technology Research Centerfor Medicinal and Functional Food, Changsha,410208, Hunan, China.
⁴ Key Laboratory of TCM Heart and Lung SyndromeDifferentiation & Medicated Diet and Dietotherapy,Changsha 410208, Hunan, China.
⁵ Key Laboratory of TCM Diagnostics of Hunan Provine, Changsha 410208, Hunan, China. Electronic address: 003561@hnucm.edu.cn.
⁶ Key Laboratory of TCM Diagnostics of Hunan Provine, Changsha 410208, Hunan, China; Hunan Engineering Technology Research Centerfor Medicinal and Functional Food, Changsha,410208, Hunan, China; Key Laboratory of TCM Heart and Lung SyndromeDifferentiation & Medicated Diet and Dietotherapy,Changsha 410208, Hunan, China. Electronic address: 003439@hnucm.edu.cn.

PMID: 40645539
DOI: 10.1016/j.jep.2025.120277

Abstract

Ethnopharmacology relevance: Di Dang decoction (DDD) is a classic prescription for blood-breaking, blood-stasis-removing and collateral-dredging in Treatise on Febrile Diseases and Synopsis of the Golden Chamber. DDD is mainly used to treat "blood stasis syndrome" and is associated with diseases such as thrombosis, ischemic stroke, and microcirculatory disorders in modern medicine. Its core mechanism is believed to improve tissue ischemia and inflammatory response by blood-breaking and blood-stasis-removing (activating blood circulation and removing stasis). However, the effects of its components and their multi-target regulatory mechanisms on cerebral ischemia-reperfusion injury (CIRI) have not been elucidated.

Aim of the study: To explore the metabolic mechanism of DDD on CIRI and its potential therapeutic effect MATERIALS AND METHODS: This study developed a rat model of middle cerebral artery occlusion-reperfusion. The efficacy and optimal dose of DDD were first detected by mNss, TTC, HE, and Nissl. Secondly, network pharmacology and non-targeted metabolomics were used to predict the treatment targets and components of Didang decoction. The specific mechanism of action of Didang decoction was finally confirmed through inflammatory factor detection, co-expression immunofluorescence, TEM, and WB.

Results: The DDD H dose group significantly reduced the neurological deficit score, reduced the volume of cerebral infarction, and restored brain tissue damage. In total, 77 components of Didang decoction were identified based on UPLC-Q-TOF-MS technology. Among these, Kaempferol, Rhein, Alizarin, and other components were associated with neuroprotection. Metabolomics screening revealed that DDD upregulated 42 metabolites and downregulated 78 metabolites. Network pharmacology predicted that core targets including AKT1 and IL6 coordinated the apoptosis-autophagy-inflammation network by modulating the PI3K/Akt signaling pathway. Animal experiments further confirmed that DDD significantly suppressed the expression of Bax, Cleaved-Caspase-3, Cytochrome C, LC3-II, Becline1, ULK1, NLRP3, and P-NF-кBp65 proteins; DDD also downregulated the levels of proinflammatory factors and activated the key nodes of the PI3K/Akt/mTOR pathway.

Conclusion: For the first time, this study systematically revealed the molecular mechanism by which Didang Decoction synergistically modulates metabolic networks and alleviates cerebral ischemia-reperfusion injury via multiple components.

Keywords: Didang decoction; PI3K/Akt/mTOR pathway; apoptosis; autophagy; cerebral ischemia-reperfusion injury; inflammation; network pharmacology; non-targeted metabolomics.