Cao Lijiao, Zhang Jing, Li Mo, Zhou Junlan, Liu Yuying, Liu Chunlan, Li Xianchan
State Key Laboratory of Natural and Biomimetic Drugs and Department of Pharmaceutical Analysis, School of Pharmaceutical Sciences, Peking University, Beijing 100191, P. R. China.
College of Life and Environmental Sciences, Minzu University of China, Beijing 100081, China.
Anal Chem. 2024 Sep 11. doi: 10.1021/acs.analchem.4c02493.
Polysaccharides, which are well-known natural macromolecules, have been recognized for their protective effects on neurons and their influence on extracellular dopamine levels in the brain. It is crucial to investigate the impact of plant polysaccharides on neurotransmission, particularly regarding the vesicular storage and exocytosis of neurotransmitters. In this study, we demonstrated the possibility of studying how the polysaccharide from Champ.(GPS) affects vesicle dopamine content and the dynamics of exocytosis in pheochromocytoma (PC12) cells using single-cell amperometry (SCA) and intracellular vesicle impact electrochemical cytometry (IVIEC). Our results unambiguously demonstrate that GPS effectively enhances vesicular neurotransmitter content and alters the dynamics of exocytosis, favoring a smaller fraction of content released in exocytotic release, thereby inducing the partial release mode. These significant effects are attributed to GPS's efficient elevation of calcium influx, significant alteration in the composition of exocytosis-related membrane lipids, and enhancement of free radical scavenging ability. These findings not only establish GPS as a promising candidate for preventive or therapeutic interventions against neurodegenerative disorders but also reiterate the importance of screening native neurologic drugs with single-vesicle electrochemical approaches, the combination of SCA and IVIEC, from a neurotransmitter-centric perspective.
多糖是众所周知的天然大分子,其对神经元的保护作用以及对大脑细胞外多巴胺水平的影响已得到认可。研究植物多糖对神经传递的影响至关重要,特别是在神经递质的囊泡储存和胞吐作用方面。在本研究中,我们证明了利用单细胞安培法(SCA)和细胞内囊泡冲击电化学细胞术(IVIEC)研究来自Champ.的多糖(GPS)如何影响嗜铬细胞瘤(PC12)细胞中囊泡多巴胺含量和胞吐动力学的可能性。我们的结果明确表明,GPS有效地提高了囊泡神经递质含量并改变了胞吐动力学,有利于在胞吐释放中释放较小比例的内容物,从而诱导部分释放模式。这些显著效果归因于GPS有效提高钙内流、显著改变与胞吐相关的膜脂组成以及增强自由基清除能力。这些发现不仅确立了GPS作为预防或治疗神经退行性疾病的有前途的候选者,还重申了从以神经递质为中心的角度,采用单囊泡电化学方法(SCA和IVIEC的组合)筛选天然神经药物的重要性。