Cystic fibrosis (CF) is a severe and diffuse recessive genetic disease due to defects of the CF Transmembrane conductance Regulator (CFTR) gene. CF affects several organs, with the chronic pulmonary disease being the major cause of reduction of the quality and expectancy of life. The hallmark of CF lung disease is chronic infection generally sustained by the gram-negative bacterium Pseudomonas aeruginosa and excessive lung inflammation with a huge infiltrate of neutrophils in the bronchial lumen, mainly due to the release of the chemokine interleukin IL-8. The identification of innovative drugs, to reduce the excessive lung inflammation in CF patients, is considered a therapeutic target to prevent the progressive lung tissue deterioration. Extracts originated from olive processing waste and obtained by a new Molecular Imprinting procedure, have been investigated to select pure compounds able to reduce IL-8 expression in human bronchial epithelial cells (IB3-1), 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 understand whether the different active principles were able to inhibit the binding between transcription factor (TF) NF-kB and DNA consensus sequences. Among different identified active principles, three derivatives were selected, apigenin, oleuropein and cyanidin chloride which have shown remarkable activity in inhibiting TF/DNA complexes. NF-kB is known to be a very interesting target molecule for the design of anti-tumor, pro-apoptotic and anti-inflammatory drugs; these natural derivatives have proved to be excellent lead compounds for the inhibition of NF-kB-p50 biological activity. At last, we demonstrated that apigenin and cyanidin chloride are able to modulate the expression of the NF-kB regulated IL-8 gene in CF IB3-1 cells, while oleuropein showed no effect in regulating the expression of the gene IL-8.
Gene Expression modulation of pro-inflammatory IL-8 in cystic fibrosis cells by active principles of olive extract
LAMPRONTI, Ilaria;VERTUANI, Silvia;MANFREDINI, Stefano;GAMBARI, Roberto
2012
Abstract
Cystic fibrosis (CF) is a severe and diffuse recessive genetic disease due to defects of the CF Transmembrane conductance Regulator (CFTR) gene. CF affects several organs, with the chronic pulmonary disease being the major cause of reduction of the quality and expectancy of life. The hallmark of CF lung disease is chronic infection generally sustained by the gram-negative bacterium Pseudomonas aeruginosa and excessive lung inflammation with a huge infiltrate of neutrophils in the bronchial lumen, mainly due to the release of the chemokine interleukin IL-8. The identification of innovative drugs, to reduce the excessive lung inflammation in CF patients, is considered a therapeutic target to prevent the progressive lung tissue deterioration. Extracts originated from olive processing waste and obtained by a new Molecular Imprinting procedure, have been investigated to select pure compounds able to reduce IL-8 expression in human bronchial epithelial cells (IB3-1), 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 understand whether the different active principles were able to inhibit the binding between transcription factor (TF) NF-kB and DNA consensus sequences. Among different identified active principles, three derivatives were selected, apigenin, oleuropein and cyanidin chloride which have shown remarkable activity in inhibiting TF/DNA complexes. NF-kB is known to be a very interesting target molecule for the design of anti-tumor, pro-apoptotic and anti-inflammatory drugs; these natural derivatives have proved to be excellent lead compounds for the inhibition of NF-kB-p50 biological activity. At last, we demonstrated that apigenin and cyanidin chloride are able to modulate the expression of the NF-kB regulated IL-8 gene in CF IB3-1 cells, while oleuropein showed no effect in regulating the expression of the gene IL-8.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
