Inhibition effects of Vitex rotundifolia on inflammatory gene expression in A549 human epithelial cells.

BACKGROUND: Vitex rotundifolia has long been used in traditional medicine to treat asthma and other allergic diseases. OBJECTIVE: To evaluate the anti-inflammatory mechanisms of V rotundifolia in cultured A549 human alveolar epithelial cells. METHODS: In the present study, A549 cells were stimulated with tumor necrosis factor alpha, interleukin 4, and interleukin 1beta to induce expression of chemokines and adhesion molecules involved in eosinophil chemotaxis. The anti-inflammatory effects of V rotundifolia on stimulated A549 cells were then evaluated by analyzing eotaxin secretion and eosinophil migration. In addition, the effects of V rotundifolia on gene expression profiles in stimulated A549 cells were evaluated by oligonucleotide microarray and real-time reverse transcription-polymerase chain reaction (RTRP). RESULTS: The V rotundifolia-treated A549 cells had significantly suppressed eotaxin secretion and eosinophil migration in a dose-dependent manner. In addition, the results of the microarray analysis and RTRP revealed that inflammation-related genes and cell adhesion-related genes were down-regulated in V rotundifolia-treated A549 cells. Furthermore, several genes related to the mitogen-activated protein kinase pathway were down-regulated in V rotundifolia-treated A549 cells. CONCLUSIONS: The mechanism responsible for the effects of V rotundifolia on A549 cells is closely associated with regulation of the mitogen-activated protein kinase pathway. Thus, V rotundifolia may be useful in the treatment of asthma and other allergic diseases.

Global gene analysis of Erigeron canadensis-treated TNF-alpha-, IL-4- and IL-1beta-stimulated A549 human epithelial cells.

BACKGROUND/AIMS: This study was conducted to evaluate the anti-inflammatory mechanisms of Erigeron canadensis (EC) on the tumor necrosis factor-alpha (TNF-alpha)-, interleukin (IL)-4- and IL-1beta-induced stimulation of A549 cells. METHODS: In the present study, the anti-inflammatory effects of EC on TNF-alpha-, IL-4- and IL-1beta-induced A549 cells were determined by analyzing eotaxin secretion using ELISA. In addition, the effects of ECon gene expression profiles in stimulated A549 cells were evaluated by microarray analysis. RESULTS: Oligonucleotide microarray analysis revealed that inflammatory-related genes such as NOS1, NOS2A, IL-1beta, IL-8 and CSF2 and cell adhesion-related genes such as SELE, MMP3, VCAM1, ICAM1, ITGA7 and ITGB2 were downregulated in EC-treated A549 cells that had been pretreated with TNF-alpha, IL-4 and IL-1beta. In addition, significant decreases in Eotaxin, ICAM, VCAM and IL-8 gene expression were observed in EC-treated A549 cells. CONCLUSIONS: EC has an anti-inflammatory effect in stimulated A549 cells. Microarray-based genomic survey is a high-throughput approach that is suitable for the evaluation of gene expression in cell lines that have been treated with EC.

Inhibitory effects of Schizandrae Fructus on eotaxin secretion in A549 human epithelial cells and eosinophil migration.

Eosinophilia have been implicated in a broad range of diseases, most notably allergic conditions (e.g. asthma, rhinitis and atopic dermatitis) and inflammatory diseases. These diseases are characterized by an accumulation of eosinophils in the affected tissue. Defining the mechanisms that control the recruitment of eosinophil is fundamental to understanding how these diseases progress and identifying a novel target for drug therapy. Accordingly, this study was conducted to evaluate the regulatory effect of Schizandrae Fructus (SF) on the expression of eotaxin, an eosinophil-specific chemokine released in respiratory epithelium following allergic stimulation, as well as its effects on eosinophil migration. To accomplish this, human epithelial lung cells (A549 cell) were stimulated with a combination of TNF-alpha (100ng/ml) and IL-4 (100ng/ml) for 24h. The cells were then restimulated with TNF-alpha (100ng/ml) and IL-1beta (10ng/ml) to induce the expression of chemokines and adhesion molecules involved in eosinophil chemotaxis for another 24h. Next, the samples were treated with various concentrations of Schizandrae Fructus (SF) (1, 10, 100, 1000microg/ml) or one of the major constituents of SF, schizandrin (0.1, 1, 10, 100microg/ml), after which following inhibition effect assay was performed triplicates in three independence. The levels of eotaxin in secreted proteins were suppressed significantly by SF (100 and 1000microg/ml, p<0.01) and schizandrin (10 and 100microg/ml, p<0.01). In addition, SF (1, 10, 100 and 1000microg/ml) decreased mRNA expression levels in A549 cells significantly (p<0.01). Eosinophil recruitment to lung epithelial cells was also reduced by SF, which indicates that eotaxin plays a role in eosinophil recruitment. Furthermore, treatment with SF suppressed the expression of another chemokine, IL-8 (0.1 and 1microg/ml SF, p<0.01), as well as intercellular adhesion molecule-1 (10 and 100microg/ml SF, p<0.01) and vascular cell adhesion molecule-1 (0.1 and 1microg/ml SF, p<0.05), which are all related to eosinophil migration. Taken together, these findings indicate that SF may be a desirable medicinal plant for the treatment of allergic diseases.

Inhibition effects of Moutan Cortex Radicis on secretion of eotaxin in A549 human epithelial cells and eosinophil migration.

UNLABELLED: Eosinophils have been implicated in a broad range of diseases, most notably allergic conditions (e.g. asthma, rhinitis and atopic dermatitis) and inflammatory diseases. These diseases are characterized by an accumulation of eosinophils in the tissue. Defining the mechanisms that control eosinophil recruitment is fundamental to understanding how these diseases progress and may identify a novel target for drug therapy. Eotaxin is a potent eosinophil-specific chemokine that is released in the respiratory epithelium after allergic stimulation. AIM OF THE STUDY: In this study, we determined whether Moutan Cortex Radicis (MCR), a plant extract, effects eotaxin secretion from A549 epithelial cells and eosinophil chemotaxis, and then examined the mechanism involved. MATERIALS AND METHODS: Prior to assaying MCR's effects, A549 cells were stimulated with tumor necrosis factor-alpha (TNF-alpha), interleukin-4 (IL-4) and IL-1beta to induce expression of chemokines and adhesion molecules involved in eosinophil chemotaxis. In the presence of MCR, eotaxin, regulated on activation in normal T cells expressed and secreted (RANTES), IL-8, IL-16, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) transcripts were quantitated by real-time RT-PCR. RESULTS: As a result, 0.01, 1, and 100 microg/ml of MCR treatments reduced eotaxin expression significantly and 0.01, 0.1, 1, 10, and 100 microg/ml of MCR reduced significantly eotaxin secretion. In addition, MCR treatment significantly inhibited eosinophil migration toward A549 medium. And 100 microg/ml of MCR suppressed the activated of nuclear factor (NF)-kappaB. CONCLUSIONS: These findings indicate that suppressed eotaxin secretion by MCR treatment is due to the inhibition of NF-kappaB activation. Therefore, MCR might be of therapeutic value in treating asthma.