Department of Life Science and Biotechnology, Laboratory of Medicinal and Molecular Biotechnology, University of Ferrara, Via Fossato di Mortara 74, 44121 Ferrara, Italy.
Evid Based Complement Alternat Med. 2013;2013:960603. doi: 10.1155/2013/960603. Epub 2013 Jul 7.
A persistent recruitment of neutrophils in the bronchi of cystic fibrosis (CF) patients contributes to aggravate the airway tissue damage, suggesting the importance of modulating the expression of chemokines, including IL-8 during the management of the CF patients. Polyphenols rich extracts derived from waste water from olive mill, obtained by a molecular imprinting approach, have been investigated in order to discover compounds able to reduce IL-8 expression in human bronchial epithelial cells (IB3-1 cells), derived from a CF patient with a ΔF508/W1282X mutant genotype and stimulated with TNF-alpha. Initially, electrophoretic mobility shift assays (EMSAs) were performed to determine whether the different active principles were able to inhibit the binding between transcription factor (TF) NF-kappaB and DNA consensus sequences. Among different representative active principles present in the extract, three compounds were selected, apigenin, oleuropein, and cyanidin chloride, which displayed remarkable activity in inhibiting NF-kappaB/DNA complexes. Utilizing TNF-alpha-treated IB3-1 cells as experimental model system, we demonstrated that apigenin and cyanidin chloride are able to modulate the expression of the NF-kappaB-regulated IL-8 gene, while oleuropein showed no effect in regulating the expression of the gene IL-8.
在囊性纤维化 (CF) 患者的支气管中持续招募中性粒细胞会加重气道组织损伤，这表明在 CF 患者的管理中调节趋化因子（包括 IL-8）表达的重要性。通过分子印迹方法从橄榄压榨废水获得的富含多酚的提取物已被研究，以发现能够降低 TNF-α刺激的 CF 患者ΔF508/W1282X 突变基因型衍生的人支气管上皮细胞 (IB3-1 细胞) 中 IL-8 表达的化合物。最初，进行电泳迁移率变动分析 (EMSA) 以确定不同的活性成分是否能够抑制转录因子 (TF) NF-κB 与 DNA 共有序列之间的结合。在提取物中存在的不同代表性活性成分中，选择了三种化合物，即芹菜素、橄榄苦苷和氯化飞燕草素，它们在抑制 NF-κB/DNA 复合物方面表现出显著的活性。利用 TNF-α 处理的 IB3-1 细胞作为实验模型系统，我们证明了芹菜素和氯化飞燕草素能够调节 NF-κB 调节的 IL-8 基因的表达，而橄榄苦苷则没有调节 IL-8 基因表达的作用。
