Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University 150081, 157 Baojian Road, Nangang District, China.
Nutr Metab Cardiovasc Dis. 2012 Mar;22(3):215-22. doi: 10.1016/j.numecd.2010.06.013. Epub 2010 Sep 28.
Chronic inflammation plays pivotal roles in both cancer and cardiovascular diseases. A large body of evidence suggests that high intake of cruciferous vegetables is closely related with low risk of these disorders. However, the underlying mechanisms of protection are not fully understood. The aim of this study is to test the protective effects of an isothiocyanate sulphoraphane on inflammatory injury and related regulation pathways in cultured endothelial cells.
The expressions of adhesion molecules were determined by TaqMan real-time polymerase chain reaction (PCR) and Western blot analysis. Nuclear factor-kappa B (NF-кB) translocation was detected by immunofluorescent hybridisation. Other proteins were measured by Western blot analysis. The results demonstrated that sulphoraphane significantly suppresses the expression of intercellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 stimulated by lipopolysaccharide (LPS) both at the transcriptional and translational levels. In addition, sulphoraphane inhibited the translocation of NF-кB into the nucleus. Sulphoraphane decreased the phosphorylation of extra-cellular signal-regulated kinase (ERK), Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK), while further blockade and activation using individually specific agents confirm that p38 MAPK and JNK are mainly involved. Interestingly, sulphoraphane down-regulated Toll-like receptor (TLR)-4, a receptor of LPS located on the membrane. In addition, MyD88, an effector downstream TLR-4 signal pathway was subsequently attenuated.
Taken all together, adhesion molecules are confirmed to be the novel targets of sulphoraphane in preventing inflammatory insult to endothelial cells. Sulphoraphane suppressed TLR-4 followed by MyD88 and downstream factors such as p38 MAPK and JNK, ultimately blocking NF-кB translocation and the subsequent expression of adhesion molecules. These data suggested a novel inflammatory pathway mediated by sulphoraphane.
慢性炎症在癌症和心血管疾病中都起着关键作用。大量证据表明，大量摄入十字花科蔬菜与这些疾病的低风险密切相关。然而，其保护机制尚未完全阐明。本研究旨在检测异硫氰酸酯萝卜硫素（sulforaphane）对培养的内皮细胞炎症损伤及相关调节途径的保护作用。
采用 TaqMan 实时聚合酶链反应（PCR）和 Western blot 分析检测黏附分子的表达。通过免疫荧光杂交检测核因子-κB（NF-κB）易位。Western blot 分析检测其他蛋白。结果表明，萝卜硫素可显著抑制脂多糖（LPS）刺激的细胞间黏附分子（ICAM）-1和血管细胞黏附分子（VCAM）-1的表达，无论是在转录水平还是在翻译水平上。此外，萝卜硫素抑制 NF-κB 向核内易位。萝卜硫素降低细胞外信号调节激酶（ERK）、Jun N-末端激酶（JNK）和 p38 丝裂原活化蛋白激酶（p38 MAPK）的磷酸化，而使用单独的特异性药物进一步阻断和激活证实 p38 MAPK 和 JNK 主要参与其中。有趣的是，萝卜硫素下调了位于细胞膜上 LPS 的受体 Toll 样受体（TLR）-4。此外，TLR-4 信号通路的下游效应子 MyD88 随后也被减弱。
综上所述，黏附分子被证实是萝卜硫素预防内皮细胞炎症损伤的新靶点。萝卜硫素通过抑制 TLR-4 及其下游因子如 p38 MAPK 和 JNK，最终阻断 NF-κB 易位及随后黏附分子的表达，从而抑制炎症反应。这些数据提示了萝卜硫素介导的一条新的炎症通路。
