Department of Food Science and Technology, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea.
J Ethnopharmacol. 2010 Oct 5;131(3):550-4. doi: 10.1016/j.jep.2010.07.030. Epub 2010 Jul 23.
This study is to elucidate the involvement of anti-inflammatory heme oxygenase-1 (HO-1) in the inhibitory activity of a Chrysanthemum boreale Makino (CB) extract on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages.
Cell viability and NO assay were performed. In addition, iNOS expression was detected by Western blotting and real-time PCR. HO-1 expression was also evaluated by Western blotting, and blocking HO-1 activity on NO production was performed.
The CB extract at the highest concentration (100 μg/ml) significantly inhibited NO production by approximately 90% and suppressed iNOS protein expression by approximately 84.8% compared to LPS-stimulated cells. Furthermore, the CB extract (100 μg/ml) inhibited iNOS mRNA expression in a concentration-dependant manner and suppressed iNOS mRNA expression by 94.8%. The CB extract induced the expression of HO-1 in a dose-dependent manner, and blocking HO-1 activity abolished the inhibitory effects of the CB extract. Moreover, the addition of carbon monoxide such as tricarbonyl dichlororuthenium (II) dimmer (RuCO), a byproduct derived from heme degradation, mimicked the inhibitory action of low concentrations of CB extract.
These results suggest that a CB extract has potent anti-inflammatory activity in RAW264.7 macrophages involving the induction of HO-1.
本研究旨在阐明抗炎血红素加氧酶-1（HO-1）在 Chrysanthemum boreale Makino（CB）提取物抑制脂多糖（LPS）刺激的 RAW264.7 巨噬细胞一氧化氮（NO）产生和诱导型一氧化氮合酶（iNOS）表达中的作用。
进行细胞活力和 NO 测定。此外，通过 Western blot 和实时 PCR 检测 iNOS 表达。通过 Western blot 评估 HO-1 表达，并阻断 HO-1 对 NO 产生的活性。
与 LPS 刺激的细胞相比，CB 提取物在最高浓度（100 μg/ml）时可显著抑制约 90%的 NO 产生，并抑制约 84.8%的 iNOS 蛋白表达。此外，CB 提取物（100 μg/ml）以浓度依赖的方式抑制 iNOS mRNA 表达，并抑制 iNOS mRNA 表达达 94.8%。CB 提取物以剂量依赖的方式诱导 HO-1 的表达，阻断 HO-1 活性可消除 CB 提取物的抑制作用。此外，添加一氧化碳，如三羰基二氯钌（II）二聚体（RuCO），血红素降解的副产物，模拟低浓度 CB 提取物的抑制作用。
这些结果表明，CB 提取物在 RAW264.7 巨噬细胞中具有强大的抗炎活性，涉及 HO-1 的诱导。
