Aslim Belma, Nigdelioglu Dolanbay Serap, Baran Sahra Setenay
Gazi University, Faculty of Science, Department of Biology, Ankara 06500, Turkey.
Gazi University, Faculty of Science, Department of Biology, Ankara 06500, Turkey.
Comput Biol Chem. 2024 Oct;112:108144. doi: 10.1016/j.compbiolchem.2024.108144. Epub 2024 Jul 9.
Alzheimer's disease (AD) is characterized by neuronal loss due to hyperphosphorylated proteins induced by oxidative stress. AD remains a formidable challenge in the medical field, as current treatments focusing on single biomarkers have yielded limited success. Hence, there's a burgeoning interest in investigating novel compounds that can target mechanisms, offering alternative therapeutic approaches. The aim of this study is to investigate the effects of allocryptopine, an isoquinoline alkaloid, on mechanisms related to AD in order to develop alternative treatment strategies. In this study, the in vitro AD cell model was obtained by inducing nerve growth factor (NGF)-differentiated PC12 (dPC12) cells to oxidative stress with HO, and also the effect mechanism of different allocryptopine concentrations on the in vitro AD cell model was studied. The treatments' antioxidative effects at the ROS level and their regulation of the cell cycle were assessed through flow cytometry, while their anti-apoptotic effects were evaluated using both flow cytometry and qRT-PCR. Additionally, the phosphorylation levels of Akt, GSK-3β, and tau proteins were analyzed via western blot, and the interactions between Akt, GSK-3β, CDK5 proteins, and allocryptopine were demonstrated through molecular docking. Our study's conclusive results revealed that allocryptopine effectively suppressed intracellular ROS levels, while simultaneously enhancing the Akt/GSK-3β signaling pathway by increasing p-Akt and p-GSK-3β proteins. This mechanism played a critical role in inhibiting neural cell apoptosis and preventing tau hyperphosphorylation. Moreover, allocryptopine demonstrated its ability to regulate the G1/S cell cycle progression, leading to cell cycle arrest in the G1 phase, and facilitating cellular repair mechanisms, potentially contributing to the suppression of neural apoptosis. The in silico results of allocryptopine were shown to docking with the cyclin-dependent kinase 5 (CDK 5) playing a role in tau phosphorylation Akt and GSK-3β from target proteins. Therefore, the in silico study results supported the in vitro results. The results showed that allocryptopine can protect dPC12 cells from oxidative stress-induced apoptosis and hyperphosphorylation of the tau protein by regulating the Akt/GSK-3β signaling pathway. Based on these findings, it can be suggested that allocryptopine, with its ability to target biomarkers and its significant effects on AD-associated mechanisms, holds promise as a potential candidate for drug development in the treatment of AD. Further research and clinical trials are recommended in the future.
阿尔茨海默病（AD）的特征是由于氧化应激诱导的蛋白质过度磷酸化导致神经元丢失。AD在医学领域仍然是一个巨大的挑战，因为目前专注于单一生物标志物的治疗方法取得的成功有限。因此，人们对研究能够靶向相关机制的新型化合物产生了浓厚兴趣，以提供替代治疗方法。本研究的目的是研究异喹啉生物碱别隐品碱对与AD相关机制的影响，从而制定替代治疗策略。在本研究中，通过用HO诱导神经生长因子（NGF）分化的PC12（dPC12）细胞产生氧化应激来获得体外AD细胞模型，并研究了不同浓度别隐品碱对体外AD细胞模型的作用机制。通过流式细胞术评估处理在ROS水平的抗氧化作用及其对细胞周期的调节，同时使用流式细胞术和qRT-PCR评估其抗凋亡作用。此外，通过蛋白质印迹分析Akt、GSK-3β和tau蛋白的磷酸化水平，并通过分子对接证明Akt、GSK-3β、CDK5蛋白与别隐品碱之间的相互作用。我们研究的最终结果表明，别隐品碱有效抑制细胞内ROS水平，同时通过增加p-Akt和p-GSK-3β蛋白增强Akt/GSK-3β信号通路。该机制在抑制神经细胞凋亡和防止tau过度磷酸化中起关键作用。此外，别隐品碱显示出调节G1/S细胞周期进程的能力，导致细胞周期停滞在G1期，并促进细胞修复机制，可能有助于抑制神经细胞凋亡。别隐品碱的计算机模拟结果显示与在tau磷酸化中起作用的细胞周期蛋白依赖性激酶5（CDK 5）以及来自靶蛋白的Akt和GSK-3β对接。因此，计算机模拟研究结果支持了体外实验结果。结果表明，别隐品碱可通过调节Akt/GSK-3β信号通路保护dPC12细胞免受氧化应激诱导的凋亡和tau蛋白过度磷酸化。基于这些发现，可以认为别隐品碱具有靶向生物标志物的能力及其对AD相关机制的显著影响，有望成为治疗AD药物开发的潜在候选物。建议未来进行进一步的研究和临床试验。
