Chuncheon Center, Korea Basic Science Institute, Chuncheon, 24341, Republic of Korea.
Chin J Integr Med. 2021 Apr;27(4):273-279. doi: 10.1007/s11655-021-3330-9. Epub 2021 Mar 24.
To demonstrate the anti-inflammatory activity of Brassica napus L. hydrosols (BNH) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells.
Composition analysis of BNH was conducted via gas chromatography-mass spectrometry after BNH were extracted. The nitric oxide (NO) production was measured using the Griess assay. Prostaglandin E (PGE) production was evaluated with enzyme-linked immunosorbent assay. The effects of BNH on LPS-induced pro-inflammatory enzymes including inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were evaluated using Western blot analysis. Furthermore, phosphorylation of nuclear factor-kappa B (NF-κB) and nuclear translocation of NF-κB p65 were evaluated with Western blot analysis and immunofluorescence staining, respectively.
Compared with LPS-stimulated cells, BNH markedly decreased the generation of NO and PGE in LPS-stimulated RAW 264.7 cells (P<0.01 or P<0.05). Moreover, BNH inhibited protein levels of iNOS and COX-2 (P<0.01). Phosphorylation of NF-κB and nuclear translocation of NF-κB p65 was significantly inhibited by BNH (P<0.01 or P<0.05).
The anti-inflammatory activities of BNH were mediated via blockage of the NF-κB signaling pathways in LPS-stimulated RAW 264.7 cells.
展示油菜（Brassica napus L.）水提物（BNH）在脂多糖（LPS）刺激的 RAW 264.7 细胞中的抗炎活性。
在提取 BNH 后，通过气相色谱-质谱联用分析其组成。通过格里斯测定法测量一氧化氮（NO）的产生。用酶联免疫吸附试验评估前列腺素 E（PGE）的产生。使用 Western blot 分析评估 BNH 对 LPS 诱导的促炎酶（包括诱导型一氧化氮合酶（iNOS）和环氧化酶-2（COX-2））的影响。此外，通过 Western blot 分析和免疫荧光染色分别评估核因子-κB（NF-κB）的磷酸化和 NF-κB p65 的核转位。
与 LPS 刺激的细胞相比，BNH 显著降低了 LPS 刺激的 RAW 264.7 细胞中 NO 和 PGE 的产生（P<0.01 或 P<0.05）。此外，BNH 抑制了 iNOS 和 COX-2 的蛋白水平（P<0.01）。NF-κB 的磷酸化和 NF-κB p65 的核转位均被 BNH 显著抑制（P<0.01 或 P<0.05）。
BNH 的抗炎活性是通过阻断 LPS 刺激的 RAW 264.7 细胞中的 NF-κB 信号通路介导的。
