Source Paper
Ruozhi Dang, Mingyang Wang, Xinhui Li, Haiyang Wang, Lanxiang Liu et al.
Journal of Neuroinflammation • 2022
Abstract Background The inflammation and oxidative stress (OS) have been considered crucial components of the pathogenesis of depression. Edaravone (EDA), a free radical scavenger, processes strong biological activities including antioxidant, anti-inflammatory and neuroprotective properties. However, its role and potential molecular mechanisms in depression remain unclear. The present study aimed to investigate the antidepressant activity of EDA and its underlying mechanisms. Methods A chronic social defeat stress (CSDS) depression model was performed to explore whether EDA could produce antidepressant effects. Behaviors tests were carried out to examine depressive, anxiety-like and cognitive behaviors including social interaction (SI) test, sucrose preference test (SPT), open field test (OFT), elevated plus maze (EPM), novel object recognition (NOR), tail suspension test (TST) and forced swim test (FST). Hippocampal and medial prefrontal cortex (mPFC) tissues were collected for Nissl staining, immunofluorescence, targeted energy metabolomics analysis, enzyme-linked immunosorbent assay (ELISA), measurement of MDA, SOD, GSH, GSH-PX, T-AOC and transmission electron microscopy (TEM). Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) detected the Sirt1/Nrf2/HO-1/Gpx4 signaling pathway. EX527, a Sirt1 inhibitor and ML385, a Nrf2 inhibitor were injected intraperitoneally 30 min before EDA injection daily. Knockdown experiments were performed to determine the effects of Gpx4 on CSDS mice with EDA treatment by an adeno-associated virus (AAV) vector containing miRNAi (Gpx4)–EGFP infusion. Results The administrated of EDA dramatically ameliorated CSDS-induced depressive and anxiety-like behaviors. In addition, EDA notably attenuated neuronal loss, microglial activation, astrocyte dysfunction, oxidative stress damage, energy metabolism and pro-inflammatory cytokines activation in the hippocampus (Hip) and mPFC of CSDS-induced mice. Further examination indicated that the application of EDA after the CSDS model significantly increased the protein expressions of Sirt1, Nrf2, HO-1 and Gpx4 in the Hip. EX527 abolished the antidepressant effect of EDA as well as the protein levels of Nrf2, HO-1 and Gpx4. Similarly, ML385 reversed the antidepressant and anxiolytic effects of EDA via decreased expressions of HO-1 and Gpx4. In addition, Gpx4 knockdown in CSDS mice abolished EDA-generated efficacy on depressive and anxiety-like behaviors. Conclusion These findings suggest that EDA possesses potent antidepressant and anxiolytic properties through Sirt1/Nrf2/HO-1/Gpx4 axis and Gpx4-mediated ferroptosis may play a key role in this effect.
Objective: Measure social interaction and social avoidance as an indicator of depression-like behavior in a chronic social defeat stress (CSDS) depression model
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Perform chronic social defeat stress protocol to establish depression model in mice
Note: This is the baseline stress induction procedure
“A chronic social defeat stress (CSDS) depression model was performed”
Conduct social interaction test to measure social avoidance as indicator of depression-like behavior
Note: One of multiple behavioral tests performed
“Behaviors tests were carried out to examine depressive, anxiety-like and cognitive behaviors including social interaction (SI) test”
Inject EX527 (Sirt1 inhibitor) and ML385 (Nrf2 inhibitor) intraperitoneally 30 minutes before EDA injection daily
Note: Injections performed daily as part of knockdown experiments
“EX527, a Sirt1 inhibitor and ML385, a Nrf2 inhibitor were injected intraperitoneally 30 min before EDA injection daily”
Administer EDA injection daily following inhibitor injection
Note: Timing follows 30 minutes after inhibitor injection
“EX527, a Sirt1 inhibitor and ML385, a Nrf2 inhibitor were injected intraperitoneally 30 min before EDA injection daily”
Perform adeno-associated virus vector containing miRNAi (Gpx4)–EGFP infusion for knockdown experiments
Note: Knockdown experiments performed to determine effects of Gpx4 on CSDS mice with EDA treatment
“Knockdown experiments were performed to determine the effects of Gpx4 on CSDS mice with EDA treatment by an adeno-associated virus (AAV) vector containing miRNAi (Gpx4)–EGFP infusion”
Collect hippocampal and medial prefrontal cortex (mPFC) tissues for analysis
Note: Tissues collected for multiple downstream analyses
“Hippocampal and medial prefrontal cortex (mPFC) tissues were collected”
Perform Nissl staining on collected tissues
Note: Part of tissue analysis protocol
“Hippocampal and medial prefrontal cortex (mPFC) tissues were collected for Nissl staining”
Perform immunofluorescence on collected tissues
Note: Part of tissue analysis protocol
“Hippocampal and medial prefrontal cortex (mPFC) tissues were collected for Nissl staining, immunofluorescence”
Perform targeted energy metabolomics analysis on collected tissues
Note: Part of tissue analysis protocol
“targeted energy metabolomics analysis”
Perform enzyme-linked immunosorbent assay (ELISA) on collected tissues
Note: Part of tissue analysis protocol
“enzyme-linked immunosorbent assay (ELISA)”
Measure MDA, SOD, GSH, GSH-PX, and T-AOC levels in collected tissues
Note: Oxidative stress biomarkers measured
“measurement of MDA, SOD, GSH, GSH-PX, T-AOC”
Perform transmission electron microscopy (TEM) on collected tissues
Note: Ultrastructural analysis of tissues
“transmission electron microscopy (TEM)”
Perform Western blotting (WB) to detect Sirt1/Nrf2/HO-1/Gpx4 signaling pathway
Note: Protein expression analysis
“Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) detected the Sirt1/Nrf2/HO-1/Gpx4 signaling pathway”
Perform quantitative real-time polymerase chain reaction (qRT-PCR) to detect Sirt1/Nrf2/HO-1/Gpx4 signaling pathway
Note: Gene expression analysis
“Western blotting (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) detected the Sirt1/Nrf2/HO-1/Gpx4 signaling pathway”
CSDS depression model was performed on subjects
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