Hu Wenqing, He Jianghong, Wang Yu, Xu Lingchi, Zhao Ying, Hu Xinping, Shen Hongmei
Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education, Co-innovation Center of Neuroregeneration, Nantong University, Nantong, China.
Department of Bioengineering, Jacobs School of Engineering, UC San Diego, La Jolla, CA, USA.
Ann Transl Med. 2021 Feb;9(3):248. doi: 10.21037/atm-20-581.
Achyranthes bidentata polypeptides (ABPPs) are a potent intervention for excitotoxicity-related disorders such as Parkinson's disease and ischemic stroke. Previous work suggests that overstimulation of N-methyl-D-aspartate (NMDA) receptors plays a critical role in excitotoxicity, and expression of NR2 subunit variations is developmentally regulated. Our current study focused on neuroprotection of ABPPs on cultured neurons by modulation of NR2A and NR2B differentially.
Primary cultured neurons were treated with NVP-AAM077, Ro-256981, ABPPs, and then the neurons were exposed to NMDA to induce excitotoxicity. Cellular viability was detected promptly and 24-hour after exposure to NMDA by MTT assay. Patch-clamp recording was applied to evaluate the effect of ABPPs on NMDA-evoked current and the differential modulation of ABPPs on NR2A and NR2B subunits in conjunction with NVP-AAM077 and Ro-256981.
ABPPs (10 µg/mL) blocked neuronal injury by NMDA in mature cultures, and the peptides conferred neuroprotection in immature cultures unless co-applied with NVP-AAM077. Furthermore, ABPPs enhanced NMDA current in mature cultures, while decreasing NMDA current in immature cultures. On the other hand, we showed that ABPPs increased NMDA current when Ro-256981 was present and decreased NMDA current when NVP-AAM007 was present.
Neuroprotection of ABPPs on NMDA-mediated injury differentially in immature and mature cultures involves enhancement of NR2A subunits and prevention of NR2B subunits, indicating that dosage of ABPP should be considered in treatment with patients at different developmental stages.
牛膝多肽（ABPPs）是帕金森病和缺血性中风等与兴奋性毒性相关疾病的有效干预措施。先前的研究表明，N-甲基-D-天冬氨酸（NMDA）受体的过度刺激在兴奋性毒性中起关键作用，并且NR2亚基变体的表达受发育调控。我们目前的研究集中在ABPPs通过差异调节NR2A和NR2B对培养神经元的神经保护作用。
原代培养神经元用NVP-AAM077、Ro-256981、ABPPs处理，然后将神经元暴露于NMDA以诱导兴奋性毒性。通过MTT法在暴露于NMDA后立即和24小时检测细胞活力。应用膜片钳记录来评估ABPPs对NMDA诱发电流的影响以及ABPPs与NVP-AAM077和Ro-256981联合对NR2A和NR2B亚基的差异调节。
ABPPs（10μg/mL）在成熟培养物中可阻断NMDA诱导的神经元损伤，并且除非与NVP-AAM077共同应用，这些肽在未成熟培养物中具有神经保护作用。此外，ABPPs在成熟培养物中增强NMDA电流，而在未成熟培养物中降低NMDA电流。另一方面，我们表明当存在Ro-256981时ABPPs增加NMDA电流，而当存在NVP-AAM007时降低NMDA电流。
ABPPs对未成熟和成熟培养物中NMDA介导损伤的神经保护作用存在差异，涉及增强NR2A亚基和抑制NR2B亚基，表明在治疗不同发育阶段的患者时应考虑ABPPs的剂量。
