School of Life Sciences and Biopharmaceutics, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, Liaoning 110016, People's Republic of China; Department of Pharmacy, General Hospital of Northern Military Area, 83 Wenhua Road, Shenyang, Liaoning 110016, People's Republic of China.
Department of Pharmacy, General Hospital of Northern Military Area, 83 Wenhua Road, Shenyang, Liaoning 110016, People's Republic of China.
Toxicol In Vitro. 2020 Sep;67:104888. doi: 10.1016/j.tiv.2020.104888. Epub 2020 May 19.
Compound DCMQA (4, 5-O-dicaffeoyl-1-O-[4-malic acid methyl ester]-quinic acid) is a natural caffeoylquinic acid derivative isolated from Arctium lappa L. roots. Caffeoylquinic acid derivatives have been reported to possess neuroprotective effects through inhibiting oxidative stress and apoptosis in vitro. However, whether DCMQA exerts protective effects on N-methyl-D-aspartate (NMDA)-induced neurotoxicity and the underlying mechanism has not been elucidated. In this study, the results indicated that pretreatment of DCMQA prevented the loss of cell viability and attenuated the LDH leakage in SH-SY5Y cells exposed to NMDA. Hoechst 33342 staining and Annexin V-PI double staining illustrated that DCMQA suppressed NMDA-induced morphological damage and neuronal apoptosis. Moreover, DCMQA inhibited NMDA-mediated Ca influx, excessive intracellular ROS generation and loss of mitochondrial membrane potential (MMP). Western blot analysis showed that DCMQA attenuated the Bax/Bcl-2 ratio, release of cytochrome c as well as expression of caspase-9 and caspase-3. Besides, DCMQA down-regulated GluN2B-containing NMDA receptors (NMDARs) and up-regulated GluN2A-containing NMDARs, promoted the disruption of nNOS and PSD95 as well as activation of CaMK II-α. Furthermore, computational docking study indicated that DCMQA possessed a good affinity for NMDARs. These results indicated that DCMQA protects SH-SY5Y cells against NMDA-induced neuronal damage. In addition, the underlying mechanisms of DCMQA-mediated neuroprotection are associated with modulating NMDARs and disruption of nNOS-PSD95 as well as the activation of CaMK II-α.
化合物 DCMQA（4,5-O-二咖啡酰基-1-O-[4-苹果酸甲酯]-奎宁酸）是从牛蒡根中分离得到的一种天然咖啡酰奎宁酸衍生物。研究表明，咖啡酰奎宁酸衍生物通过体外抑制氧化应激和细胞凋亡，具有神经保护作用。然而，DCMQA 是否对 N-甲基-D-天冬氨酸（NMDA）诱导的神经毒性具有保护作用及其潜在机制尚未阐明。在本研究中，结果表明 DCMQA 预处理可防止 SH-SY5Y 细胞暴露于 NMDA 时细胞活力丧失和 LDH 漏出。Hoechst 33342 染色和 Annexin V-PI 双重染色表明，DCMQA 抑制 NMDA 诱导的形态损伤和神经元凋亡。此外，DCMQA 抑制 NMDA 介导的 Ca 内流、过量的细胞内 ROS 生成和线粒体膜电位（MMP）丧失。Western blot 分析表明，DCMQA 降低了 Bax/Bcl-2 比值、细胞色素 c 的释放以及 caspase-9 和 caspase-3 的表达。此外，DCMQA 下调了含有 GluN2B 的 NMDA 受体（NMDARs），上调了含有 GluN2A 的 NMDARs，促进了 nNOS 和 PSD95 的破坏以及 CaMK II-α 的激活。此外，计算对接研究表明 DCMQA 与 NMDARs 具有良好的亲和力。这些结果表明，DCMQA 可保护 SH-SY5Y 细胞免受 NMDA 诱导的神经元损伤。此外，DCMQA 介导的神经保护作用的潜在机制与调节 NMDARs 以及破坏 nNOS-PSD95 和激活 CaMK II-α 有关。
