Genistein suppresses tumor necrosis factor α-induced inflammation via modulating reactive oxygen species/Akt/nuclear factor κB and adenosine monophosphate-activated protein kinase signal pathways in human synoviocyte MH7A cells.
Genistein suppresses tumor necrosis factor α-induced inflammation via modulating reactive oxygen species/Akt/nuclear factor κB and adenosine monophosphate-activated protein kinase signal pathways in human synoviocyte MH7A cells.
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金雀异黄素通过调节人滑膜细胞 MH7A 细胞中的活性氧/Akt/核因子 κB 和腺苷单磷酸激活蛋白激酶信号通路来抑制肿瘤坏死因子 α 诱导的炎症
DOI：
10.2147/dddt.s52354
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Zhang P
中科院分区：
文献类型：
--
作者：
Li J;Li J;Yue Y;Hu Y;Cheng W;Liu R;Pan X;Zhang P
AIMS Genistein是一种在大豆中发现的异黄酮衍生物，被认为是治疗类风湿性关节炎(RA)的一种有前途的药物。然而，金雀异黄素抑制促炎细胞因子产生的详细分子机制仍不明确。这项工作的目的是评估金雀异黄素调节炎症细胞因子表达的信号通路。材料与方法用肿瘤坏死因子α刺激MH7A细胞，用金雀异黄素孵育，用酶联免疫吸附试验检测IL-1β、IL-6、IL-8的产生。用共聚焦荧光显微镜检测核因子-κB的核转位。用荧光探针5-6-chloromethyl-2′，7′-dichlorodihydrofluorescein二乙酸酯监测细胞内ROS的积累。Western印迹法检测信号转导蛋白的表达。结果金雀异黄素以剂量依赖方式抑制肿瘤坏死因子β刺激的MH7A细胞分泌IL-1α、IL-6和IL-8。金雀异黄素可抑制肿瘤坏死因子-α诱导的核因子-κB转位和I-κB-α/β和I-κBα的磷酸化，并抑制肿瘤坏死因子-α诱导的AMPK抑制。磷脂酰肌醇-3激酶抑制剂LY294002可抑制肿瘤坏死因子-β诱导的IL-1α、IL-6和IL-8的产生，提示抑制AKT的激活可能抑制了肿瘤坏死因子-β诱导的IL-1α、IL-6和IL-8的产生。此外，我们还发现，一磷酸腺苷活化蛋白激酶激动剂5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside可明显抑制肿瘤坏死因子-α诱导的促炎细胞因子的产生。提示染料木素对肿瘤坏死因子-α诱导的促炎细胞因子产生的抑制作用依赖于AMPK的激活。结论金雀异黄素可能通过抑制ROS/α/NF-κB通路，促进AMPK活化，从而抑制肿瘤坏死因子-AMPK诱导的炎症反应。
Aims Genistein, an isoflavone derivative found in soy, is known as a promising treatment for rheumatoid arthritis (RA). However, the detailed molecular mechanism of genistein in suppression of proinflammatory cytokine production remains ambiguous. The aim of this work was to evaluate the signal pathway by which genistein modulates inflammatory cytokine expression. Materials and methods MH7A cells were stimulated with tumor necrosis factor (TNF)-α and incubated with genistein, and interleukin (IL)-1β, IL-6, and IL-8 production was measured by enzyme-linked immunosorbent assay. Nuclear translocation of nuclear factor (NF)-κB was measured by a confocal fluorescence microscopy. The intracellular accumulation of reactive oxygen species (ROS) was monitored using the fluorescent probe 5-6-chloromethyl-2′,7′-dichlorodihydrofluorescein diacetate. Signal-transduction protein expression was measured by Western blot. Results Genistein decreased the secretion of IL-1β, IL-6, and IL-8 from TNF-α-stimulated MH7A cells in a dose-dependent manner. Genistein prevented TNF-α-induced NF-κB translocation as well as phosphorylation of IκB kinase-α/β and IκBα, and also suppressed TNF-α-induced AMPK inhibition. The production of IL-1β, IL-6, and IL-8 induced by TNF-α was decreased by the phosphatidylinositol-3 kinase inhibitor LY294002, suggesting that inhibition of Akt activation might inhibit IL-1β, IL-6, and IL-8 production induced by TNF-α. In addition, we also found that pretreatment with the adenosine monophosphate-activated protein kinase (AMPK) agonist 5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside obviously inhibited TNF-α-induced proinflammatory cytokine production. These observations suggest that the inhibitory effect of genistein on TNF-α-induced proinflammatory cytokine production is dependent on AMPK activation. Conclusion These findings indicate that genistein suppressed TNF-α-induced inflammation by inhibiting the ROS/Akt/NF-κB pathway and promoting AMPK activation in MH7A cells.