背景:加速神经突生长和停止神经变性是在中枢神经系统(CNS)的各种神经变性疾病或损伤中受到负调节的最关键因素。功能性食物或营养素被认为是生物活性成分的替代来源,可通过促进神经生成和突触发生来减轻各种中枢神经系统损伤。而其确切的分子机制仍未被探索。
目的:山梨科(CS)是一种流行的草本植物,其中CNS调节作用是传统上有据可查的,但对记忆增强功能的详细研究仍未探索。因此,本研究旨在分析CS乙醇水溶液(CSAE)提取物在原代海马神经元中的神经源性和突触调节。
方法:培养原代海马神经元并使其与CSAE或载体一起孵育。观察早期神经元分化,轴突和树突状乔木,和突触形成,神经元针对指定的抗体进行免疫染色,或用亲脂性染料直接染色(1,1'-双十八烷基-3,3,3',3\'-四甲基吲哚花青高氯酸盐,DiL).同时,使用蛋白质印迹来验证与载体相比CSAE的突触发生效应。此外,应用分子对接和系统药理学方法来确认CSAE在神经生成中发挥作用的可能的次级代谢产物和途径。
结果:结果表明,在30μg/mL浓度下,CSAE可以诱导神经突发生和突触发生。治疗影响早期神经元极化,轴突和树突状乔木,突触发生,通过原代神经元中的NMDARs表达和突触可塑性。CS代谢物的计算机网络药理学表明,CSAE介导的神经源性效应可能依赖于NTRK2(TrkB)介导的神经营养蛋白信号通路。的确,在与TrkB特异性抑制剂共同处理后,观察到的CSAE的神经源性活性显著降低。此外,分子对接后的结合能计算表明,其中一种CS代谢物,scoparone,对TrkB的BDNF模拟位点具有高亲和力,提示其在TrkB激活中的作用。史帕龙被发现可以增强神经生成,但与CSAE的程度不同。此外,TrkB信号相关蛋白(BCL2,CASP3,GSK3和BDNF)的表达,被发现是由scoparone调制的,受到TrkB抑制剂(ANA-12)的共同处理的显著影响。这些结果进一步表明,scoparone对神经生成的调节是TrkB依赖性的。
结论:这项研究为CS促进神经元生长和记忆功能的分子机制提供了更深入的见解,这可能意味着许多神经系统疾病的预防。
BACKGROUND: Acceleration of neurite outgrowth and halting neurodegeneration are the most critical factors that are negatively regulated in various neurodegenerative diseases or injuries in the central nervous system (CNS). Functional foods or nutrients are considered alternative sources of bioactive components to alleviate various CNS injuries by promoting neuritogenesis and synaptogenesis, while their exact molecular mechanism remains unexplored.
OBJECTIVE: Coriandrum sativum L. (CS) is one of the popular herbs in the Apiaceae family, of which CNS modulating action is a well-documented traditionally but detailed
study on memory boosting function yet remains unexplored. Consequently, this
study aims to analyze the neurogenic and synaptogenic modulation of CS aqueous ethanol (CSAE) extract in the primary hippocampal neurons.
METHODS: Primary hippocampal neurons were cultured and allowed to incubate with CSAE or vehicle. To observe the early neuronal differentiation, axonal and dendritic arborization, and synapse formation, neurons were immune-stained against indicated antibodies or stained directly with a lipophilic dye (1, 1\'-dioctadecyl-3, 3, 3\', 3\'-tetramethyl indocarbocyanine perchlorate, DiL). Meanwhile, western blot was used to validate the
synaptogenesis effect of CSAE compared to vehicle. Additionally, molecular docking and system pharmacology approaches were applied to confirm the possible secondary metabolites and pathways by which CSAE promotes neuritogenesis.
RESULTS: Results show that CSAE can induce neuritogenesis and synaptogenesis at 30 μg/mL concentration. The treatment impacts early neuronal polarization, axonal and dendritic arborization,
synaptogenesis, and synaptic plasticity via NMDARs expressions in primary neurons. In silico network pharmacology of CS metabolites show that the CSAE-mediated neurogenic effect is likely dependent on the NTRK2 (TrkB) mediated neurotrophin signaling pathway. Indeed, the observed neurogenic activity of CSAE is markedly reduced upon the co-treatment with a TrkB-specific inhibitor. Furthermore, molecular docking following binding energy calculation shows that one of the CS metabolites, scoparone, has a high affinity to bind in the BDNF mimetic binding site of TrkB, suggesting its role in TrkB activation. Scoparone was found to enhance neuritogenesis, but not to the same extent as CSAE. Moreover, the expression of TrkB signaling-related proteins (BCL2, CASP3, GSK3, and BDNF), which was found to be modulated by scoparone, was significantly affected by the co-treatment of TrkB inhibitor (ANA-12). These results further suggest that the modulation of neuritogenesis by scoparone is TrkB-dependent.
CONCLUSIONS: This
study provides deeper insights into the molecular mechanism of CS in boosting neuronal growth and memory function, which might implicate the prevention of many neurological disorders.