Abstract 1118 Gardenin A as a Neurotherapeutic Agent: Mitigating Alcohol-Induced Brain Damage and Beyond

Introduction: Alcohol is often associated with liver effects, but it impacts multiple vital organs, including the brain and the gut. Alcohol can cross the blood-brain barrier (BBB), generating stress mechanisms through pathways like alcohol dehydrogenase and cytochrome P4502E1. Excessive alcohol consumption leads to brain damage, cognitive and neuronal loss, and a decrease in brain volume, affecting white matter. Currently, no FDA-approved drugs address alcohol-induced brain damage. At present, pharmacotherapy is emerging with minimal side effects. Notably, Gardenin A (GA), a polymethoxylated flavone with high bioavailability and efficacy, has shown effectiveness against various ailments, and the ability to stimulate neuritogenesis, offering strategies for combating neurological diseases. Methods: For our study, we isolated GA from Dikamali gum (Gum resin of Gardenia resinifera Roth.) and characterized it using NMR spectrometry and High Resolution-Liquid Chromatography-Mass Spectrometry (HR-LCMS). To simulate the pathophysiological conditions of Alcohol-Induced Brain Damage, we conducted experiments using the neuronal cell line SH-SY5Y. Cell viability was assessed via the Alamar blue assay, with GA concentrations ranging from 10 to 100 μg / ml. In investigating the underlying mechanisms, we quantified Reactive Oxygen Species (ROS) generation using the dichlorofluorescein diacetate (DCFDA) assay. Furthermore, gene expression studies were carried out through Reverse Transcription-quantitative Polymerase Chain Reaction (RT-qPCR) to evaluate the modulation of pro- and anti-inflammatory genes, and those specific to tight junctions and brain function, shedding light on the relationship between GA and neuronal health. Results: GA was isolated as yellow needle-like crystals, was confirmed to be pure and authentic through characterization. In terms of cell viability assessments, GA demonstrated no significant toxicity up to a concentration of 50 μg / ml. Notably, the presence of GA corresponded to a reduction in ROS generation in comparison to the disease model. In our gene expression analysis, pro-inflammatory genes exhibited downregulation, while anti-inflammatory genes displayed upregulation. Additionally, genes related to tight junctions demonstrated increased expression levels as compared to the disease model. In the realm of brain function-related genes, Brain-Derived Neurotrophic Factor (BDNF) showed upregulation, while the pro-inflammatory gene MCP-1 exhibited downregulation. Conclusions and Future perspectives: Our study successfully uncovered the therapeutic potential of GA in alleviating alcohol-induced neuronal damage by its antioxidant, anti-inflammatory, and therapeutic qualities, leading to reduced neuro-inflammation, enhanced brain function and BBB integrity. Further mechanistic studies and clinical trials may explore the potential of GA, potentially in combination therapies to enhance treatment effectiveness. Overall, our research lays the groundwork for advancing our understanding and treatment options for alcohol-induced neurological conditions. We would like to extend our gratitude towards Lady Tata Memorial Trusts for providing a Junior Scholarship and funding for our research. We would also like to thank PD Patel Institute of Applied Sciences (CHARUSAT) and Indian Institute of Technology (IIT, Gandhinagar) for providing the high-end technology for this study to be carried out smoothly.