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首页> 外文期刊>World Journal of Gastroenterology >DA-9601, a standardized extract of Artemisia asiatica, blocks TNF-alpha-induced IL-8 and CCL20 production by inhibiting p38 kinase and NF-kappaB pathways in human gastric epithelial cells.
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DA-9601, a standardized extract of Artemisia asiatica, blocks TNF-alpha-induced IL-8 and CCL20 production by inhibiting p38 kinase and NF-kappaB pathways in human gastric epithelial cells.

机译：DA-9601是一种青蒿的标准化提取物，它通过抑制人胃上皮细胞中的p38激酶和NF-κB途径来阻断TNF-α诱导的IL-8和CCL20的产生。

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摘要

AIM: To investigate whether, or how, DA-9601, which is a new gastroprotective agent, inhibits TNF-alpha-induced inflammatory signals in gastric epithelial AGS cells. METHODS: Cell viability was determined by MTT assay. IL-8 and CCL20 promoter activities were determined by a luciferase reporter gene assay. NF-kappaB-dependent transcriptional activity was determined by I-kappaB alpha degradation, NF-kappaB p65 nuclear translocation and a luciferase activity assay. IL-8 and CCL20 gene expression and protein secretion were determined by RT-PCR and an enzyme-linked immunosorbent assay (ELISA). Total and phosphorylated forms of mitogen-activated protein kinases (MAPKs) were determined by Western blot. RESULTS: Treatment of AGS cells with DA-9601 reduced TNF-alpha-induced IL-8 and CCL20 promoter activities, as well as their gene expression and protein release. TNF-alpha also induced NF-kappaB-dependent transcriptional activity in AGS cells. In contrast, in cells treated with DA-9601, TNF-alpha-induced NF-kappaB activity was significantly blocked. Although all three MAP kinase family members were phosphorylated in response to TNF-alpha, a selective inhibitor of p38 kinase SB203580 only could inhibit both NF-kappaB-dependent transcriptional activity and IL-8 and CCL20 production, suggesting a potential link between p38 kinase and NF-kappaB-dependent pathways in AGS cells. Interestingly, DA-9601 also selectively inhibited p38 kinase phosphorylation induced by TNF-alpha. CONCLUSION: DA-9601 blocked TNF-alpha-mediated inflammatory signals by potentially modulating the p38 kinase pathway and/or a signal leading to NF-kappaB-dependent pathways in gastric epithelial cells.
机译：目的：研究新型胃保护剂DA-9601是否或如何抑制TNF-α诱导的胃上皮AGS细胞中的炎症信号。方法：采用MTT法测定细胞活力。 IL-8和CCL20启动子活性通过荧光素酶报道基因测定来确定。 NF-kappaB依赖的转录活性是通过I-kappaBα降解，NF-kappaB p65核易位和荧光素酶活性测定来确定的。通过RT-PCR和酶联免疫吸附测定（ELISA）测定IL-8和CCL20基因表达和蛋白质分泌。通过蛋白质印迹法测定促分裂原活化蛋白激酶（MAPK）的总和磷酸化形式。结果：DA-9601处理AGS细胞可降低TNF-α诱导的IL-8和CCL20启动子活性，以及​​其基因表达和蛋白质释放。 TNF-α还诱导AGS细胞中NF-κB依赖性转录活性。相反，在用DA-9601处理的细胞中，TNF-α诱导的NF-κB活性被显着阻断。尽管所有三个MAP激酶家族成员均响应TNF-α磷酸化，但p38激酶SB203580的选择性抑制剂只能抑制NF-κB依赖的转录活性以及IL-8和CCL20的产生，这表明p38激酶与AGS细胞中的NF-κB依赖性途径。有趣的是，DA-9601还选择性抑制TNF-α诱导的p38激酶磷酸化。结论：DA-9601可通过潜在地调节胃上皮细胞中的p38激酶途径和/或导致NF-κB依赖性途径的信号来阻断TNF-α介导的炎症信号。

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