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.

中文翻译：

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1-Methoxylespeflorin G11 通过抑制 ROS 产生和细胞凋亡保护 HT22 细胞免受谷氨酸诱导的细胞死亡。

本研究旨在研究 1-methoxylespeflorin G11 (MLG)（一种紫檀素）对神经元 HT22 海马细胞中谷氨酸诱导的神经毒性的神经保护作用。使用 MTT 法和显微镜分析评估 MLG 的保护作用。使用对用膜联蛋白 V/碘化丙锭探测的受损细胞进行的流式细胞术分析来研究细胞凋亡的程度。此外，使用流式细胞术通过 MitoSOXTM Red 染色评估线粒体活性氧 (ROS)。为了确定线粒体膜电位，用四甲基罗丹明和 JC-1 进行染色，然后进行流式细胞术。结果表明，MLG 通过抑制细胞内 ROS 的产生和线粒体功能障碍来减弱 HT22 细胞中谷氨酸诱导的细胞凋亡。此外，MLG 通过上调 Bcl-2 和下调裂解的 PARP-1、AIF 和磷酸化 MAPK 级联，阻止 HT22 细胞中谷氨酸诱导的凋亡途径。此外，MLG 处理诱导 HT22 细胞中的 HO-1 表达。这些结果表明，MLG 通过抑制氧化应激和细胞凋亡，对神经元 HT22 细胞中谷氨酸诱导的神经毒性表现出神经保护作用。