2. Academic Validation
  3. Anti-inflammatory effect of Danhong injection through inhibition of GSDMD-mediated pyroptosis

Anti-inflammatory effect of Danhong injection through inhibition of GSDMD-mediated pyroptosis

  • Phytomedicine. 2023 Mar 5;113:154743. doi: 10.1016/j.phymed.2023.154743.
Yujuan Li 1,  Zizhuo Tu 2,  Feng Chen 2,  Xingbo Yang 2,  Ruhua Deng 2,  Fanghua Su 2,  Zhiyang Cheng 2,  Shengxuan Li 2,  Sang-Bing Ong 3,  Dandan Wang 2,  Yang Chen 2,  Yaozu Xiang 4
Affiliations

Affiliations

  • 1 Shanghai Key Laboratory of Anesthesiology and Brain Functional Modulation, School of Life Sciences and Technology, Shanghai Fourth People's Hospital, Tongji University, Shanghai 200434, China; Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
  • 2 Shanghai Key Laboratory of Anesthesiology and Brain Functional Modulation, School of Life Sciences and Technology, Shanghai Fourth People's Hospital, Tongji University, Shanghai 200434, China.
  • 3 Department of Medicine and Therapeutics, Faculty of Medicine, Chinese University of Hong Kong (CUHK), 999077, Hong Kong SAR; Centre for Cardiovascular Genomics and Medicine (CCGM), Lui Che Woo Institute Innovative Medicine, Chinese University of Hong Kong (CUHK), 999077, Hong Kong SAR.
  • 4 Shanghai Key Laboratory of Anesthesiology and Brain Functional Modulation, School of Life Sciences and Technology, Shanghai Fourth People's Hospital, Tongji University, Shanghai 200434, China; Institute of Biophysics, Chinese Academy of Science, Beijing 100101, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China; Key Laboratory of Arrhythmias of the Ministry of Education of China, School of Life Sciences and Technology, Shanghai East Hospital, Tongji University, Shanghai 20092, China. Electronic address: [email protected].
PMID: 36893672 DOI: 10.1016/j.phymed.2023.154743
Abstract

Background: Pyroptosis is an inflammatory form of cell death that has been implicated in various infectious and non-infectious diseases. Gasdermin family proteins are the key executors of pyroptotic cell death, thus they are considered as novel therapeutic targets for inflammatory diseases. However, only limited gasdermin specific inhibitors have been identified to date. Traditional Chinese medicines have been applied in clinic for centuries and exhibit potential in anti-inflammation and anti-pyroptosis. We attempted to find candidate Chinese botanical drugs which specifically target gasdermin D (GSDMD) and inhibit Pyroptosis.

Methods: In this study, we performed high-throughput screening using a botanical drug library to identify Pyroptosis specific inhibitors. The assay was based on a cell Pyroptosis model induced by lipopolysaccharides (LPS) and nigericin. Cell Pyroptosis levels were then evaluated by cell cytotoxicity assay, propidium iodide (PI) staining and immunoblotting. We then overexpressed GSDMD-N in cell lines to investigate the direct inhibitory effect of the drug to GSDMD-N oligomerization. Mass spectrometry studies were applied to identify the active components of the botanical drug. Finally, a mouse model of sepsis and a mouse model of diabetic myocardial infarction were constructed to verify the protective effect of the drug in disease models of inflammation.

Results: High-throughput screening identified Danhong injection (DHI) as a Pyroptosis Inhibitor. DHI remarkably inhibited pyroptotic cell death in a murine macrophage cell line and bone marrow-derived macrophages. Molecular assays demonstrated the direct blockade of GSDMD-N oligomerization and pore formation by DHI. Mass spectrometry studies identified the major active components of DHI, and further activity assays revealed salvianolic acid E (SAE) as the most potent molecule among these components, and SAE has a strong binding affinity to mouse GSDMD Cys192. We further demonstrated the protective effects of DHI in mouse sepsis and mouse myocardial infarction with type 2 diabetes.

Conclusion: These findings provide new insights for drug development from Chinese herbal medicine like DHI against diabetic myocardial injury and sepsis through blocking GSDMD-mediated macrophage Pyroptosis.

Keywords

DHI; GSDMD; Pyroptosis; TCM.

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