Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
Department of Ophthalmology, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83301, Taiwan.
Int J Mol Sci. 2023 Apr 14;24(8):7300. doi: 10.3390/ijms24087300.
Microglia-associated neuroinflammation is recognized as a critical factor in the pathogenesis of neurodegenerative diseases; however, there is no effective treatment for the blockage of neurodegenerative disease progression. In this study, the effect of nordalbergin, a coumarin isolated from the wood bark of , on lipopolysaccharide (LPS)-induced inflammatory responses was investigated using murine microglial BV2 cells. Cell viability was assessed using the MTT assay, whereas nitric oxide (NO) production was analyzed using the Griess reagent. Secretion of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) was detected by the ELISA. The expression of inducible NO synthase (iNOS), cyclooxygenase (COX)-2, mitogen-activated protein kinases (MAPKs) and NLRP3 inflammasome-related proteins was assessed by Western blot. The production of mitochondrial reactive oxygen species (ROS) and intracellular ROS was detected using flow cytometry. Our experimental results indicated that nordalbergin ≤20 µM suppressed NO, IL-6, TNF-α and IL-1β production; decreased iNOS and COX-2 expression; inhibited MAPKs activation; attenuated NLRP3 inflammasome activation; and reduced both intracellular and mitochondrial ROS production by LPS-stimulated BV2 cells in a dose-dependent manner. These results demonstrate that nordalbergin exhibits anti-inflammatory and anti-oxidative activities through inhibiting MAPK signaling pathway, NLRP3 inflammasome activation and ROS production, suggesting that nordalbergin might have the potential to inhibit neurodegenerative disease progression.
小胶质细胞相关的神经炎症被认为是神经退行性疾病发病机制中的一个关键因素；然而，目前尚无阻止神经退行性疾病进展的有效治疗方法。在这项研究中，研究了从白皮松木材树皮中分离得到的香豆素 nordalbergin 对脂多糖（LPS）诱导的炎症反应的影响，采用小鼠小胶质细胞 BV2 细胞作为研究模型。通过 MTT 检测法评估细胞活力，通过 Griess 试剂分析一氧化氮（NO）的产生。通过 ELISA 检测白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）的分泌。通过 Western blot 检测诱导型一氧化氮合酶（iNOS）、环氧化酶（COX）-2、丝裂原活化蛋白激酶（MAPKs）和 NLRP3 炎性体相关蛋白的表达。通过流式细胞术检测线粒体活性氧（ROS）和细胞内 ROS 的产生。我们的实验结果表明，20µM 以下的 nordalbergin 抑制了 LPS 刺激的 BV2 细胞中 NO、IL-6、TNF-α和 IL-1β的产生；降低了 iNOS 和 COX-2 的表达；抑制了 MAPKs 的激活；减弱了 NLRP3 炎性体的激活；并减少了 LPS 刺激的 BV2 细胞中细胞内和线粒体 ROS 的产生，呈剂量依赖性。这些结果表明， nordalbergin 通过抑制 MAPK 信号通路、NLRP3 炎性体激活和 ROS 产生发挥抗炎和抗氧化作用，提示 nordalbergin 可能具有抑制神经退行性疾病进展的潜力。
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