Department of Anatomy, Dongguk University College of Medicine, Gyeongju, 38066, Republic of Korea.
Department of Biochemistry and Molecular Biology, Bangladesh Agricultural University, Mymensingh, 2202, Bangladesh.
Sci Rep. 2019 Dec 12;9(1):18919. doi: 10.1038/s41598-019-55364-8.
Potentiating neuritogenesis through pharmacological intervention might hold therapeutic promise in neurodegenerative disorders and acute brain injury. Here, we investigated the novel neuritogenic potentials of a steroidal chlorohydrin, 3β, 6β-dichloro-5-hydroxy-5α-cholestane (hereafter, SCH) and the change in cellular proteome to gain insight into the underlying mechanism of its neurotrophic activity in hippocampal neurons. Morphometric analysis showed that SCH promoted early neuronal differentiation, dendritic arborization and axonal maturation. Proteomic and bioinformatic analysis revealed that SCH induced upregulation of several proteins, including those associated with neuronal differentiation and development. Immunocytochemical data further indicates that SCH-treated neurons showed upregulation of Hnrnpa2b1 and Map1b, validating their proteomic profiles. In addition, a protein-protein interaction network analysis identified TrkA as a potential target connecting most of the upregulated proteins. The neurite outgrowth effect of SCH was suppressed by TrkA inhibitor, GW441756, verifying TrkA-dependent activity of SCH, which further supports the connection of TrkA with the upregulated proteins. Also, the computational analysis revealed that SCH interacts with the NGF-binding domain of TrkA through Phe327 and Asn355. Collectively, our findings provide evidence that SCH promotes neuronal development via upregulating TrkA-signaling proteins and suggest that SCH could be a promising therapeutic agent in the prevention and treatment of neurodegenerative disorders.
通过药理学干预增强神经突生成能力在神经退行性疾病和急性脑损伤中具有治疗潜力。在这里，我们研究了一种甾体氯醇，3β，6β-二氯-5-羟基-5α-胆甾烷（以下简称 SCH）的新型神经突生成潜力，并研究了细胞蛋白质组的变化，以深入了解其在海马神经元中的神经营养活性的潜在机制。形态计量学分析表明，SCH 促进了早期神经元分化、树突分枝和轴突成熟。蛋白质组学和生物信息学分析显示，SCH 诱导了几种蛋白质的上调，包括与神经元分化和发育相关的蛋白质。免疫细胞化学数据进一步表明，SCH 处理的神经元上调了 Hnrnpa2b1 和 Map1b，验证了它们的蛋白质组谱。此外，蛋白质-蛋白质相互作用网络分析表明，TrkA 是连接大多数上调蛋白的潜在靶点。SCH 的神经突生长作用被 TrkA 抑制剂 GW441756 抑制，验证了 SCH 的 TrkA 依赖性活性，这进一步支持了 TrkA 与上调蛋白的联系。此外，计算分析表明，SCH 通过 Phe327 和 Asn355 与 TrkA 的 NGF 结合域相互作用。总之，我们的研究结果提供了证据表明，SCH 通过上调 TrkA 信号蛋白促进神经元发育，并表明 SCH 可能成为预防和治疗神经退行性疾病的有希望的治疗剂。
