College of Pharmacy and Research Institute of Pharmaceutical Science and Technology, Ajou University, Suwon 16499, Republic of Korea.
Biometrology Group, Korea Research Institute of Standards and Science (KRISS), Daejeon 34113, Republic of Korea.
J Microbiol Biotechnol. 2021 Feb 28;31(2):217-225. doi: 10.4014/jmb.2011.11032.
This study aimed to investigate the neuroprotective effects of 1-methoxylespeflorin G11 (MLG), a pterocarpan, against glutamate-induced neurotoxicity in neuronal HT22 hippocampal cells. The protective effects of MLG were evaluated using MTT assay and microscopic analysis. The extent of apoptosis was studied using flow cytometric analysis performed on the damaged cells probed with annexin V/propidium iodide. Moreover, mitochondrial reactive oxygen species (ROS) were assessed using flow cytometry through MitoSOXTM Red staining. To determine mitochondrial membrane potential, staining with tetramethylrhodamine and JC-1 was performed followed by flow cytometry. The results demonstrated that MLG attenuates glutamate-induced apoptosis in HT22 cells by inhibiting intracellular ROS generation and mitochondrial dysfunction. Additionally, MLG prevented glutamate-induced apoptotic pathway in HT22 cells through upregulation of Bcl-2 and downregulation of cleaved PARP-1, AIF, and phosphorylated MAPK cascades. In addition, MLG treatment induced HO-1 expression in HT22 cells. These results suggested that MLG exhibits neuroprotective effects against glutamate-induced neurotoxicity in neuronal HT22 cells by inhibiting oxidative stress and apoptosis.
本研究旨在探讨紫檀芪 G11（MLG）对谷氨酸诱导的 HT22 海马神经元细胞毒性的神经保护作用。通过 MTT 分析和显微镜分析评估 MLG 的保护作用。使用流式细胞术分析用 Annexin V/碘化丙啶标记的受损细胞，研究细胞凋亡的程度。此外，通过 MitoSOXTM Red 染色用流式细胞术评估线粒体活性氧（ROS）。通过用四甲基罗丹明和 JC-1 染色，然后进行流式细胞术来确定线粒体膜电位。结果表明，MLG 通过抑制细胞内 ROS 生成和线粒体功能障碍来减轻 HT22 细胞中谷氨酸诱导的凋亡。此外，MLG 通过上调 Bcl-2 和下调 cleaved PARP-1、AIF 和磷酸化 MAPK 级联来防止 HT22 细胞中谷氨酸诱导的凋亡途径。此外，MLG 处理诱导 HT22 细胞中 HO-1 的表达。这些结果表明，MLG 通过抑制氧化应激和凋亡，对谷氨酸诱导的 HT22 神经元细胞毒性具有神经保护作用。
