In Situ Amplification of 5-Lipoxygenase and 5-Lipoxygenase-Activating Protein in Allergic Airway Inflammation and Inhibition by Leukotriene Blockade

摘要: Leukotrienes are important mediators of the eosinophilic influx and mucus hypersecretion in lungs a murine model asthma. We used situ PCR this human asthma to detect lung mRNA for 5-lipoxygenase (5-LO) 5-LO-activating protein (FLAP), key proteins necessary leukotriene synthesis. Lung tissue was obtained on day 28 from mice treated with i.p. (days 0 14) intranasal 14, 25, 26, 27) OVA or saline. After fixation, sections underwent protease- RNase-free DNase digestion, before RT-PCR using target-specific cDNA amplification. 5-LO FLAP-specific visualized by digoxigenin detection system, positive cells were analyzed morphometry. associated primarily eosinophils alveolar macrophages airways pulmonary blood vessels OVA-sensitized/challenged mice. FLAP expression increased per-cell basis OVA-treated compared saline controls. Pulmonary vessel endothelial also 5-LO, mRNA, protein. inhibition significantly decreased inflammatory cells. These studies demonstrate marked increase pathway allergic response an immunomodulatory effect blockade decrease gene expression.