| 曾赛丽，吴 广，谢 莉，张秀峰，谭小武，何振华，游晓星，赵兰华，曾焱华.芹菜素激活c-Src/NADPH氧化酶/ROS通路诱导HO-1.[J].中南医学科学杂志.,2015,43(4):369-373. |
| 芹菜素激活c-Src/NADPH氧化酶/ROS通路诱导HO-1 |
| Apigenin Induces HO-1 Expression by c-Src/NADPH Oxidase/ROS to Protect Against Excessive Cytokines Secretion in Murine Macrophages |
| 投稿时间：2015-04-22 |
| DOI： |
| 中文关键词: 芹菜素 血红素氧合酶-1 小鼠巨噬细胞 |
| 英文关键词:Apigenin Heme oxygenase-1 Murine macrophages |
| 基金项目:湖南省卫生厅资助项目（B2011—067）. |
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| 中文摘要: |
| 目的 观察芹菜素对脂多糖（LPS）诱导小鼠巨噬细胞分泌细胞因子的影响，并探讨其分子机制。 方法 体外培养小鼠巨噬细胞RAW 264.7，用不同浓度的芹菜素处理后，加入LPS刺激0～16 h。Western blot检测血红素氧合酶-1（HO-1）的表达以及c-Src和NADPH氧化酶p47phox亚基的磷酸化；荧光探针H2DCFDA检测活性氧（ROS）的产生；ELISA检测TNF-α和IL-6的产生。 结果 芹菜素作用RAW 264.7细胞30 min即可诱导c-Src和NADPH氧化酶亚基p47phox磷酸化。采用50 μmol/L c-Src抑制剂PP1预处理细胞后，p47phox磷酸化水平明显降低。同时，芹菜素能上调RAW264.7细胞内ROS的含量，而NADPH氧化酶抑制剂DPI可抑制其产生。芹菜素也能诱导RAW264.7细胞表达HO-1，而采用PP1、DPI或ROS抑制剂N-乙酰半胱氨酸（NAC）预处理细胞后，HO-1表达水平明显降低。此外，芹菜素处理能下调LPS诱导RAW264.7细胞分泌TNF-α和IL-6，而采用siRNA沉默HO-1表达后，能在一定程度上降低芹菜素对细胞因子的抑制效应。 结论 芹菜素可能通过激活c-Src/NADPH氧化酶/ROS通路诱导HO-1表达，从而抑制小鼠巨噬细胞过度分泌细胞因子。 |
| 英文摘要: |
| Objective To investigate the effect and the mechanism of Apigenin on lipopolysaccharides （LPS）-induced cytokines production in murine macrophages. Methods The murine macrophage cell line RAW 264.7 cells were cultured in vitro,and were treated with different concentration of Apigenin followed by LPS stimulation.Expression of heme oxygenase-1 （HO-1）,phosphorylation of c-Src and p47phox,the subunit of NADPH oxidase were detected by Western blotThe intracellular formation of reactive oxygen species （ROS） was detected by using the fluorescent probe H2DCFDASecretion of TNF-α and IL-6 were detected by ELISA. Results Western blot indicated that phosphorylated c-Src and p47phox were induced by apigenin after 30min of incubation.While 50 μmol/L of PP1,an inhibitor of c-Src,could significantly inhibit p47phox phosphorylation.In addition,Apigenin could also induce RAW264.7 cells accumulation of ROS,and this effect could be abrogated by the NAPDH oxidase inhibitor,DPI.HO-1 could be induced by Apigenin,and pretreatment of PP1,DPI and the ROS scavenger N-acetyl-cysteine （NAC） could block HO-1 production.Furthermore,apigenin could inhibit LPS-induced secretion of TNF-α and IL-6,and transfection of HO-1 siRNA could antagonize this action. Conclusion Apigenin can induce HO-1 expression by c-Src/NADPH oxidase/ROS to protect against excessive cytokines secretion in murine macrophages. |
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