Abstract 180: Brain Tissue Metabolomics Study Supports the Use of Multi-Drug Cocktail Approach for Neuroprotection After Asphyxial Cardiac Arrest in Rats

Background: We have recently shown that using a multi-drug cocktail therapy targeting multiple pathways simultaneously could be a novel therapeutic approach to reduce ischemic brain injury following asphyxial cardiac arrest (CA) and resuscitation. To further validate the affected pathways using a multi-drug cocktail, we aimed to investigate changes in brain tissue metabolites after asphyxial CA and cardiopulmonary resuscitation (CPR) in rats.

Methods: The multi-drug cocktail formulation included ten neuroprotective drugs (CoQ10, Cyclosporin A, Edaravone, Metformin, N- acetylcysteine, Poloxamer 188, SS-31, Sulbutiamine, Vitamin C, and Zoniporide) focusing on distinct metabolic pathways of ischemia-reperfusion injury. Male Sprague-Dawley rats underwent 12 min of asphyxial-CA followed by CPR and received either a multi-drug cocktail or a vehicle (n=6) immediately after resuscitation. At 2 hr post-resuscitation rats were euthanized, and the whole-brain tissue was processed for mass spectrometry for metabolomic analysis. Data were collected both in +ve and -ve ion modes and the collected data were analyzed using metaboanalyst 5.0.

Results: A total of 139 metabolites were detected in +ve ion mode whereas 132 metabolites were detected in -ve ion mode. While combining and removing a duplicate number of metabolites a total of 234 metabolites were analyzed. Our principal component analysis (PCA) data showed a shift of overall metabolites towards sham groups with multi-drug cocktail treatment after 12 min CA. Out of 234 metabolites 40 were significantly increased and 19 were significantly decreased between cocktail and vehicle-treated groups (Suggested by Volcano Plot, p=0.05, fold Change =1). A total of 15 pathways were significantly impacted between cocktail and vehicle-treated groups (p= 0.05) suggesting the broad effect of a multi-drug cocktail therapy after asphyxial CA and resuscitation

Conclusion: This study represents an innovative attempt that employs metabolomics research to elucidate the effects of a multi-drug cocktail in a severe CA rodent model, uncovering a notable alteration in the overall metabolic profile of the brain and suggesting an important role of multi-drug cocktails in ischemic injury.