College of Life and Health Sciences, Northeastern University, Shenyang, 110169, P. R. China.
Key Laboratory of Data Analytics and Optimization for Smart Industry, Northeastern University, Ministry of Education, Shenyang, 110169, P. R. China.
Mol Nutr Food Res. 2019 Oct;63(20):e1801380. doi: 10.1002/mnfr.201801380. Epub 2019 Aug 16.
The anti-neuroinflammatory effect of a novel quinolyl-substituted analogue of resveratrol (RV01) on lipopolysaccharide (LPS)-induced microglial activation is investigated, as well as the possible underlying mechanisms.
Cell viability is measured using an MTT assay. Nitric oxide (NO) release is determined by nitrite assay. The interaction between RV01 and inducible nitric oxide synthase (iNOS) is studied using molecular docking. Free radical scavenging activity and reactive oxygen species (ROS) production are determined by DPPH reduction assay and DCFH-DA assay. Pretreatment with RV01 (1-30 µm) prior to LPS (1 µg mL ) stimulation decreased NO release and iNOS expression without observable cytotoxicity. RV01 reduced the mRNA levels and secretion of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). RV01 also inhibited LPS-induced ROS production and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation. Furthermore, RV01 decreases the protein expression of toll-like receptor 4 (TLR4) and inhibits the LPS-induced activation of the mitogen-activated protein kinase (MAPK) and nuclear transcription factor-κB (NF-κB) signaling pathways. Additionally, conditioned medium from microglia co-treated with LPS and RV01 alleviates the death of SH-SY5Y cells induced by conditioned medium from activated N9 microglial cells. Lastly, a mouse neuroinflammation model is further used to confirm the effect of RV01 in vivo.
These results show that RV01 suppresses microglia-mediated neuroinflammation and protects neurons from inflammatory damage, which indicates that RV01 has great potential as a nutritional preventive strategy for neuroinflammation-related diseases.
本研究旨在探讨一种新型的白藜芦醇（RV01）喹喔啉取代物类似物对脂多糖（LPS）诱导的小胶质细胞活化的抗神经炎症作用，以及可能的潜在机制。
采用 MTT 法测定细胞活力，通过亚硝酸盐测定法测定一氧化氮（NO）释放量，利用分子对接研究 RV01 与诱导型一氧化氮合酶（iNOS）的相互作用，通过 DPPH 还原法和 DCFH-DA 法测定自由基清除活性和活性氧（ROS）的产生。结果显示，在 LPS（1μg/mL）刺激前用 RV01（1-30µM）预处理可降低 NO 释放和 iNOS 表达，而无明显细胞毒性。RV01 降低了肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的 mRNA 水平和分泌。RV01 还抑制了 LPS 诱导的 ROS 产生和烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶的激活。此外，RV01 降低了 Toll 样受体 4（TLR4）的蛋白表达，并抑制了 LPS 诱导的丝裂原活化蛋白激酶（MAPK）和核转录因子-κB（NF-κB）信号通路的激活。此外，用 LPS 和 RV01 共同处理小胶质细胞的条件培养基可减轻激活的 N9 小胶质细胞条件培养基诱导的 SH-SY5Y 细胞死亡。最后，进一步使用小鼠神经炎症模型证实了 RV01 的体内作用。
这些结果表明，RV01 可抑制小胶质细胞介导的神经炎症，保护神经元免受炎症损伤，这表明 RV01 作为神经炎症相关疾病的营养预防策略具有很大的潜力。
