Wang Zeyuan, Wu Lehao, Fu Dongmei, Zhang Yan, Zhang Chunzhi
School of Biological Engineering, Dalian Polytechnic University, Dalian 116034, China.
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China.
Foods. 2022 Jun 9;11(12):1689. doi: 10.3390/foods11121689.
The objective of our present work was to explore the possible enhanced anti-neuroinflammatory ability of fermented hemp seed in lipopolysaccharide (LPS)-stimulated N9 microglial cells and elucidate its underlying mechanism. The water extract of hemp seed was fermented by . LPS-stimulated N9 microglial cells were employed for the inflammatory cell model. The release of nitric oxide (NO) was determined by Griess assay. The cytokines and inflammatory mediator expression were measured by qPCR and ELISA. The phosphorylated key signaling proteins, including nuclear factor-κB (NF-κB), mitogen-activated protein kinases (MAPKs), and phosphatidylinositol 3-kinase (PI3K/Akt), were quantified by western blot analysis. The production of intracellular reactive oxygen species (ROS) was measured by DCFH oxidation. Fermented hemp seed (FHS) reduced NO production by downregulating inducible nitric oxide synthase (iNOS) expression in LPS-stimulated N9 microglial cells. FHS treatment decreased LPS-stimulated expression of inflammatory cytokines either on mRNA or protein levels. Moreover, FHS inhibited LPS-stimulated phosphorylation of NF-κB, MAPKs, and PI3K/Akt signaling pathways. Furthermore, FHS significantly reduced the ROS production in the cells. It was concluded that FHS exerted its anti-neuroinflammatory activities by suppressing ROS production, thus inhibiting NF-κB, MAPKs, and PI3K/Akt activation, consequently decreasing the expression levels of inflammatory mediators and cytokines.
我们目前工作的目的是探讨发酵大麻籽在脂多糖（LPS）刺激的N9小胶质细胞中可能增强的抗神经炎症能力，并阐明其潜在机制。大麻籽水提取物由……发酵。采用LPS刺激的N9小胶质细胞作为炎症细胞模型。通过Griess法测定一氧化氮（NO）的释放。通过qPCR和ELISA检测细胞因子和炎症介质的表达。通过蛋白质印迹分析对包括核因子-κB（NF-κB）、丝裂原活化蛋白激酶（MAPKs）和磷脂酰肌醇3-激酶（PI3K/Akt）在内的磷酸化关键信号蛋白进行定量。通过DCFH氧化测量细胞内活性氧（ROS）的产生。发酵大麻籽（FHS）通过下调LPS刺激的N9小胶质细胞中诱导型一氧化氮合酶（iNOS）的表达来减少NO的产生。FHS处理在mRNA或蛋白质水平上均降低了LPS刺激的炎症细胞因子的表达。此外，FHS抑制了LPS刺激的NF-κB、MAPKs和PI3K/Akt信号通路的磷酸化。此外，FHS显著降低了细胞中的ROS产生。得出的结论是，FHS通过抑制ROS产生发挥其抗神经炎症活性，从而抑制NF-κB、MAPKs和PI3K/Akt的激活，进而降低炎症介质和细胞因子的表达水平。
