Department of Anatomy and Cell Biology, RWTH Aachen University, Wendlingweg 2, 52074 Aachen, Germany.
Inflamm Res. 2010 Jun;59(6):443-50. doi: 10.1007/s00011-009-0116-5. Epub 2009 Nov 19.
The aim of this study was to investigate the signal transduction pathways involved in sulforaphane (SF) mediated inhibition of the inflammatory response to lipopolysaccharide (LPS). Additionally, we investigated the effects of SF and LPS on the activity of Nrf2.
Primary rat microglia and the murine microglia cell line BV2 were used.
Cells were treated with LPS with or without SF.
Cell viability was measured via WST-assay. Real-time RT-PCR was performed to analyze cytokine mRNA levels. The nitric oxide (NO) release was measured in LPS-stimulated microglia. The induction of various signal transduction pathways and Nrf2 was determined by Western blotting. NF-kappaB and AP-1 activation was measured by dual luciferase assay.
We showed that SF attenuates the LPS-induced increase of IL-1beta, IL-6, and TNF-alpha expression in microglia. In addition, SF significantly decreases the NO in a concentration-dependent manner. SF inhibits LPS-stimulated ERK1/2 and JNK phosphorylation and thereby inhibits the LPS-induced activation of NF-kappaB- and activator protein-1 (AP-1). Moreover, SF and LPS together are able to induce Nrf2 activation.
We showed that SF, and also LPS by itself, are able to activate the cell's defence against oxidative and electrophilic stress. We conclude that SF could be a candidate agent for anti-inflammatory treatment of the central nervous system.
本研究旨在探讨萝卜硫素（SF）抑制脂多糖（LPS）诱导的炎症反应所涉及的信号转导通路。此外，我们还研究了 SF 和 LPS 对 Nrf2 活性的影响。
原代大鼠小胶质细胞和小鼠小胶质细胞系 BV2 用于本研究。
用 LPS 或 LPS 加 SF 处理细胞。
通过 WST 分析检测细胞活力。通过实时 RT-PCR 分析细胞因子 mRNA 水平。测量 LPS 刺激的小胶质细胞中一氧化氮（NO）的释放。通过 Western blot 检测各种信号转导通路和 Nrf2 的诱导。通过双荧光素酶测定法测量 NF-κB 和 AP-1 的激活。
我们表明 SF 可减弱 LPS 诱导的小胶质细胞中白细胞介素-1β、白细胞介素-6 和肿瘤坏死因子-α表达的增加。此外，SF 还可浓度依赖性地显著降低 NO。SF 抑制 LPS 刺激的 ERK1/2 和 JNK 磷酸化，从而抑制 LPS 诱导的 NF-κB 和激活蛋白-1（AP-1）的激活。此外，SF 和 LPS 可共同诱导 Nrf2 激活。
我们表明 SF 以及 LPS 本身能够激活细胞对氧化和亲电应激的防御机制。我们得出结论，SF 可能是治疗中枢神经系统炎症的候选药物。
