Flavonoids from Inula britannica reduces oxidative stress through inhibiting expression and phosphorylation of p47phox in VSMCs
Flavonoids from Inula britannica reduces oxidative stress through inhibiting expression and phosphorylation of p47phox in VSMCs
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DOI：
10.3109/13880209.2010.550055
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发表时间：
2011-06
影响因子：
3.8
通讯作者：
Hong-bing Zhang;J. Wen;Jing Zhang;Sui‐bing Miao;Guo-yan Ma;Yan-Yan Wang-Yan;Bin Zheng;Mei Han
中科院分区：
文献类型：
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作者：
Hong-bing Zhang;J. Wen;Jing Zhang;Sui‐bing Miao;Guo-yan Ma;Yan-Yan Wang-Yan;Bin Zheng;Mei Han
英文名：Inula britanica Linn.（菊科）是一种传统的中草药，已被用于治疗支气管炎和炎症。从其花中提取的总黄酮提取物（TFEs）在体内能抑制球囊损伤诱导的新生内膜形成。目的：探讨TFE在体外抑制氧化应激及炎症反应的机制。材料与方法：用不同浓度的TFE对体外培养的大鼠血管平滑肌细胞（VSMCs）进行氧化应激处理。荧光染色法检测超氧阴离子（O2 stec −）的产生，同时测定丙二醛（MDA）和超氧化物歧化酶（SOD）的水平。采用酶联免疫吸附试验（ELISA）检测肿瘤坏死因子-α（TNF-α）水平，逆转录-PCR和western blot检测p47 phox基因表达活性，免疫沉淀法检测p47 phox磷酸化水平。结果如下：TFE可抑制H2 O2诱导的VSMCs产生O2 stec −，减少TNF-α的分泌;升高培养液中SOD活性，与槲皮素作用相似;降低培养液中MDA含量。与H2 O2对照组相比，TFE预处理可降低VSMCs中p47 phox的mRNA和蛋白表达水平，甚至降低p47 phox的磷酸化水平。讨论与结论：这些结果表明，TFE能够减轻氧化应激的产生和随后的炎症反应，通过阻止p47 phox的过度表达和激活，以及增加VSMCs分泌TNF-α。
Context: Inula britanica Linn. (Compositae) is a traditional Chinese medicinal herb that has been used to treat bronchitis and inflammation. The total flavonoid extracts (TFEs) isolated from its flowers can inhibit neointimal formation induced by balloon injury in vivo. Objective: To investigate the mechanism by which TFE suppresses oxidative stress generation and the subsequent inflammation response in vitro. Materials and methods: The cultured vascular smooth muscle cells (VSMCs) form rats were exposed to oxidative stress following pretreatment with or without TFE at different concentration. Then, fluorescence staining was used to detect superoxide anion (O2˙−) production, and the lever of maleic dialdehyde (MDA) and superoxide dismutase (SOD) was measured at the same time. Furthermore, tumor necrosis factor-α (TNF-α) was measured by enzyme linked immunosorbent assay (ELISA), reverse transcription-PCR and western blot were performed to detect the expression activity of p47phox gene, and immunoprecipitation was used to test the level of p47phox phosphorylation. Results: TFE inhibited the production of O2˙− induced by H2O2 in VSMCs, with decrease in secretion of TNF-α; elevated the activity of SOD in the medium, similar to the effect of quercetin; reduced the level of MDA in culture medium of VSMCs. The pretreatment with TFE resulted in decrease the level of p47phox mRNA and protein, and even p47phox phosphorylation in VSMCs, compared with H2O2 control. Discussion and conclusion: These findings demonstrate that TFE is capable of attenuating the oxidative stress generation and the subsequent inflammation response via preventing the overexpression and activation of p47phox and the increased TNF-α secretion in VSMCs in vitro.