Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.
H.E.J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi 75270, Pakistan.
Brain Res. 2025 Jan 1;1846:149291. doi: 10.1016/j.brainres.2024.149291. Epub 2024 Oct 22.
Coordinated proliferation and differentiation of neural stem cells (NSCs) results in continuous neurogenesis. The present study provides novel insights into the Notch intracellular signaling in neuronal cell proliferation, maintenance, migration, and differentiation regulated by naturally based Quinic acid (QA) in primary hippocampal cell culture. Further, this study might help in the discovery and development of lead molecules that can overcome the challenges in the treatment of neurodegenerative diseases. The growth supporting effect of QA was studied using Alamar Blue assay. The migratory potential of QA was evaluated using scratch assay. The in vitro HO-induced oxidative stress model was used to upregulate neuronal survival after QA treatment. The RT-qPCR and immunocytochemical analysis were performed for selected markers of Notch signaling to determine the proliferation, differentiation, and maintenance of NSCs at gene and molecular levels. The Mash1 and Ngn2 are the upstream proneural genes of the Notch pathway which were included to evaluate the differentiation of NSCs into mature neurons after treatment with QA. Furthermore, regarding the role of QA in maintaining the pool of NPCs, we used Notch1 and Hes1 markers for proliferation analysis. Also, secondary neuronal markers i.e. Pax6, PCNA, and Mcm2 were included in this study and their gene expression analysis was analyzed following treatment with QA. Based on the study's results, we suggest that naturally based QA can promote the growth and differentiation of neonatal NSCs residing in hippocampal regions into neuronal lineage. Therefore, we propose that the neurogenic potential of QA can be employed to prevent and treat neurodegenerative diseases.
神经干细胞（NSCs）的协调增殖和分化导致持续的神经发生。本研究提供了关于 Notch 细胞内信号在海马原代细胞培养中受天然奎尼酸（QA）调节的神经元细胞增殖、维持、迁移和分化的新见解。此外，这项研究可能有助于发现和开发克服神经退行性疾病治疗挑战的先导分子。使用 Alamar Blue 测定法研究 QA 的生长支持作用。使用划痕测定法评估 QA 的迁移潜力。体外 HO 诱导的氧化应激模型用于上调 QA 处理后的神经元存活。进行 RT-qPCR 和免疫细胞化学分析，以确定 Notch 信号的选定标记物，以在基因和分子水平上确定 NSCs 的增殖、分化和维持。Mash1 和 Ngn2 是 Notch 途径的上游神经前体细胞基因，包括在内，以评估 QA 处理后 NSCs 向成熟神经元的分化。此外，关于 QA 在维持 NPC 池中的作用，我们使用 Notch1 和 Hes1 标记物进行增殖分析。此外，本研究还包括二次神经元标记物 Pax6、PCNA 和 Mcm2，并对其基因表达进行了分析。根据研究结果，我们认为天然 QA 可以促进海马区新生 NSCs 的生长和分化为神经元谱系。因此，我们提出 QA 的神经发生潜力可用于预防和治疗神经退行性疾病。
