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Updated: May 24, 2025
Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
Published on: May 21, 2018
Cordycepin attenuates NLRP3/Caspase-1/GSDMD-mediated LPS-induced macrophage pyroptosis
1Center for Genomic and Personalized Medicine, Guangxi key Laboratory for Genomic and Personalized Medicine, Guangxi Collaborative Innovation Center for Genomic and Personalized Medicine, University Engineering Research Center of Digital Medicine and Healthcare, Guangxi Medical University, Nanning, Guangxi, China.
Abstract:
Pyroptosis, a form of programmed cell death driven by the NLRP3 inflammasome, is a key contributor to inflammation in various diseases. This study aimed to investigate the anti-inflammatory mechanisms of cordycepin, focusing on its role in macrophage pyroptosis. Molecular docking analysis was performed to evaluate the binding affinity of cordycepin to key pyroptosis-related proteins, including NLRP3, Caspase-1, and GSDMD. RAW264.7 cells were pre-treated with cordycepin to assess its effects on pyroptosis. Key measurements included reactive oxygen species (ROS) levels, xanthine oxidase (XO) activity, and the expression of NLRP3, Caspase-1, and GSDMD. Additionally, lactate dehydrogenase (LDH) release, interleukin (IL)-1β and IL-18 levels in the culture supernatant, and macrophage cell death rates were evaluated using Hoechst 33342/PI dual staining. The results demonstrated that cordycepin exhibits strong binding affinity for NLRP3, Caspase-1, and GSDMD. Cordycepin pre-treatment significantly reduced ROS levels and XO activity, inhibited the expression of NLRP3, cleaved-Caspase-1, and cleaved-GSDMD, and decreased pyroptosis-associated inflammatory cytokines IL-1β and IL-18, along with Caspase-1 activity. Furthermore, cordycepin reduced the macrophage pyroptosis rate. In conclusion, cordycepin inhibits macrophage pyroptosis by reducing XO activity, suppressing ROS production, and regulating the expression of key molecules in the NLRP3/Caspase-1/GSDMD pathway. These findings provide a strong experimental basis for the potential development of cordycepin as a novel anti-inflammatory agent.
Insights
Cordycepin effectively inhibits macrophage pyroptosis, a key inflammatory process, by targeting the NLRP3 inflammasome pathway. This natural compound reduces inflammatory markers and cell death, showing potential as a novel anti-inflammatory agent.
Area of Science:
- Immunology
- Pharmacology
- Cell Biology
Background:
- Pyroptosis, a critical form of programmed cell death, significantly contributes to inflammation in numerous diseases.
- The NLRP3 inflammasome is a central driver of pyroptosis and associated inflammatory responses.
Purpose of the Study:
- To investigate the anti-inflammatory mechanisms of cordycepin, specifically its effects on macrophage pyroptosis.
- To evaluate cordycepin's binding affinity to key pyroptosis-related proteins and its impact on cellular inflammatory pathways.
Main Methods:
- Molecular docking was employed to assess cordycepin's binding to NLRP3, Caspase-1, and GSDMD.
- RAW264.7 macrophages were treated with cordycepin to measure reactive oxygen species (ROS), xanthine oxidase (XO) activity, and pyroptosis markers.
- Assays included LDH release, IL-1β and IL-18 quantification, and cell death assessment via dual staining.
Main Results:
- Cordycepin demonstrated strong binding affinity to NLRP3, Caspase-1, and GSDMD.
- Pre-treatment with cordycepin significantly reduced ROS levels, XO activity, and the expression of NLRP3, cleaved-Caspase-1, and cleaved-GSDMD.
- Cordycepin inhibited Caspase-1 activity, decreased IL-1β and IL-18 levels, and reduced overall macrophage pyroptosis rates.
Conclusions:
- Cordycepin effectively suppresses macrophage pyroptosis through mechanisms involving reduced XO activity and ROS production.
- Cordycepin modulates the NLRP3/Caspase-1/GSDMD pathway, highlighting its potential as a novel anti-inflammatory therapeutic agent.
