Department of Biology, Faculty of Basic Sciences, University of Guilan, Rasht, 4193833697, Iran.
Sci Rep. 2024 Nov 21;14(1):28824. doi: 10.1038/s41598-024-80102-0.
Cholinergic treatments with an emphasis on M1 muscarinic acetylcholine receptor (mAChR) agonists as potential modulating agents are a new approach in Alzheimer's disease (AD) therapy. In previous research, we designed and characterized novel thiazolidine-2,4-dione (TZD)-derived compounds that possess anti-AD properties and enhance the expression of mAChRM1 in rats. This study evaluated a novel orthosteric agonist of mAChRM1 from related pathways that has shown promising anti-Alzheimer's disease activity. PC12 cells were exposed to various concentrations of TZ4M before they were exposed to scopolamine (3 µM). Immunocytochemistry and western blot analyses revealed that TZ4M increased the expression of mAChRM1 in differentiated cells induced by scopolamine-treated PC12 cells. The results showed that TZ4M (3 and 5 µM) markedly upregulated PKC and ChAT protein expression, and the cells were significantly protected against increased ROS levels followed by neuronal cell loss, as evidenced by the MTT assay. TUNEL staining indicated that TZ4M impeded the shaping of apoptotic bodies. Analysis of the amino acid sequences of the ligand-protein binding site indicated that TZ4M is bound to the orthosteric site (acetylcholine site). This study revealed that TZ4M, a derivative of TZD, effectively protects against scopolamine-induced damage. TZ4M, a novel mACRM1 orthosteric agonist, is promising for treating AD.
胆碱能治疗强调 M1 毒蕈碱乙酰胆碱受体 (mAChR) 激动剂作为潜在的调节因子，是阿尔茨海默病 (AD) 治疗的新方法。在之前的研究中，我们设计并表征了具有抗 AD 特性的新型噻唑烷-2,4-二酮 (TZD) 衍生化合物，并增强了大鼠中 mAChRM1 的表达。这项研究评估了一种新型的 mAChRM1 变构激动剂，它来自相关途径，表现出有希望的抗阿尔茨海默病活性。PC12 细胞在暴露于东莨菪碱 (3 µM) 之前，先暴露于不同浓度的 TZ4M 下。免疫细胞化学和 Western blot 分析显示，TZ4M 增加了东莨菪碱处理的 PC12 细胞诱导的分化细胞中 mAChRM1 的表达。结果表明，TZ4M (3 和 5 µM) 显著上调了 PKC 和 ChAT 蛋白的表达，并且细胞对增加的 ROS 水平引起的神经元细胞丢失具有明显的保护作用，这一点通过 MTT 测定得到证实。TUNEL 染色表明 TZ4M 阻碍了凋亡小体的形成。对配体-蛋白结合位点的氨基酸序列进行分析表明，TZ4M 与变构结合位点（乙酰胆碱结合位点）结合。这项研究表明，TZ4M，一种 TZD 的衍生物，有效地抵抗了东莨菪碱引起的损伤。TZ4M，一种新型的 mAChRM1 变构激动剂，有望用于治疗 AD。
