Title : In vitro investigation of neuroprotective effects: Unveiling the impact of Aβ25-35-induced oxidative stress on SH-SY5Y cells and the therapeutic potential of phytochemicals
Abstract:
Alzheimer’s disease (AD) is intricately linked to oxidative stress induced by the accumulation of amyloid beta (Aβ) peptide, disrupting cellular functionality. This comprehensive study investigates the multifaceted impacts of Aβ25-35-induced oxidative stress on human neuroblastoma SH-SY5Y cells and explores the protective potential of phytochemicals ferulic acid (FA) and Ginkgolide B (GB), both individually and in combination. The evaluation encompasses diverse parameters, including cell viability, reactive oxygen species (ROS) production, oxidative DNA base damage, and repair capabilities, with a specific focus on the base excision repair (BER) pathway's key enzyme, apurinic/apyrimidinic Endonuclease 1 (APE1). Our findings unveil the profound implications of Aβ25-35 treatment, demonstrating diminished cell viability, escalated ROS production, and heightened ROS-mediated DNA damage. The study delves into the cellular repair mechanism through the BER pathway, revealing substantial alterations in the expression of the APE1 enzyme following Aβ25-35 exposure. Remarkably, our investigation introduces a strategic pre-treatment approach with FA, GB, and their combined formulation, administered 3 hours before Aβ25-35 exposure for 24 hours. This approach yields remarkable enhancements, including augmented cell viability, reduced ROS production, significant amelioration of mitochondrial DNA (mtDNA) base damage, and heightened translational expression of APE1 across diverse cellular compartments, including cytosolic, nuclear, mitochondrial, and exosomal compartments. Notably, we emphasize the importance of revealing the proteome of exosomes, shedding light on their crucial role in mediating a wide range of effects. Exosomes, identified as carriers of secreted APE1, display improved endonuclease activity within the exosomes released by cells. This activity contributes to the rescue of nearby SH-SY5Y cells from oxidative stress damage induced by Aβ25-35, which was found to improve by the pre-treatment of phytochemicals 3 hrs before Aβ25-35. Further proteomic analysis of exosomes isolated from differently treated SH-SY5Y cells, utilizing Triple-ToF, uncovered distinct patterns. Exosomes from phytochemical-treated cells, considered a healthy microenvironment, supported physiological cell health. In contrast, exosomes from Aβ25-35-treated cells, representing a diseased/pathophysiological microenvironment, delivered oxidative and inflammatory signals, potentially harming neighboring cells. Furthermore, the application of phytochemicals, recognized as neuroprotective agents against Aβ, induces a significant shift in the cellular microenvironment, leading to a notable alteration in neuronal cell physiology. The observed distinctions in exosomal proteomes underscore their pivotal role in mediating signals that influence cellular health. This integrated approach provides valuable insights into developing therapeutic strategies for Alzheimer's disease.
Audience Take Away Notes:
- The audience can leverage the findings from this research in innovative ways, particularly in the context of therapeutic development. Given that exosomes can cross the blood-brain barrier, researchers and clinicians can design experiments to isolate exosomes and encapsulate the identified neuroprotective phytochemicals. This encapsulation strategy could serve as a targeted delivery system, potentially enhancing the efficacy of these compounds in mitigating the hallmarks of diseases, including Alzheimer's. Moreover, manipulating exosomes to carry specific cargo or signals allows for the design of experiments involving the injection of these modified exosomes into mouse models. This approach could offer insights into the modulation of cellular responses and the potential therapeutic impact in vivo. Furthermore, researchers could extend these concepts to design human-based studies, exploring the feasibility of using exosome-based interventions to reduce disease markers in clinical settings. Overall, the practical applications encompass targeted drug delivery strategies, in vivo model experiments, and the potential translation of findings into human-focused therapeutic studies.
- Professionals in the fields of neuroscience, pharmacology, and healthcare can leverage the study's findings to advance their understanding of Alzheimer's disease mechanisms. The insights into Aβ-induced oxidative stress and the protective effects of phytochemical pre-treatment offer potential avenues for developing novel therapeutic interventions. Healthcare practitioners may find implications for patient care strategies, emphasizing the importance of early intervention using specific phytochemical combinations. Researchers can use this knowledge to guide further studies in drug development, targeting APE1 and the BER pathway for potential Alzheimer's treatments. Overall, this research equips professionals with actionable insights that may inform future approaches to Alzheimer's disease management.
- This research serves as a valuable resource for faculty across neuroscience, pharmacology, and related fields. It provides a foundational understanding of Alzheimer's disease mechanisms, offering opportunities for expansion in areas such as oxidative stress, neuroprotection, and cellular signaling. The study's focus on phytochemical interventions also holds potential for drug development. Faculty can integrate these insights into teaching materials, fostering a comprehensive understanding of cellular impacts and therapeutic strategies among students. The emphasis on interdisciplinary aspects, particularly in exosome research, encourages collaborative exploration across diverse academic domains.
