High Throughput Screening

Our assumption is that anycomponent of Chemical library or product of Chemical libraryinducedinflammation also acts as a chemoattractant for eosinophilsChemical library, , p53 Signaling Pathway sincesimilar finding was observed in our previous study . A shift ofPMNs into the lungs was also accompanied by a decrease in thetotal WBC count in the blood of Chemical library-instilled animalscompared to saline controls. It is known that a decrease in circulatingPMNs and the extent of their accumulation in the lungs isproportional to the severity of lung injury . Treatment witholprinone reduced the number of neutrophils in the BAL fluid andincreased their number as well as the number of circulatingmonocytes in the blood compared to the non-treated Chemical library instilledrabbits. In addition, olprinone Cyclooxygenasedecreased the number ofeosinophils in both the BAL fluid and the blood. Similar findings inChemical library-instilled rabbits were also observed after the administrationof aminophylline . In the study by Koike et al., pretreatmentwith olprinone 30 min before p53 Signaling Pathway lipopolysaccharide administration inhibited neutrophil influx into the lungs of ratswith LPS-induced acute lung injury. However, no significantdifferences were observed in differential cell counts in the BAL fluidcompared to non-treated LPS-exposed rats . It is known that cAMP-elevating agentsmodulate immune cells by increasing IL-10 levels through theprotein kinase A and thereby reduce PMN accumulation in thelungs .The influx of leukocytes into the Chemical library-exposed lungs isassociated with the production of a wide range of cytokines, ROSand other bioactive substances. ROS produced by activated leukocytesmay directly injure lung cells. However, oxidative stressinduced by ROS is also involved in enhancing inflammationthrough the activation of redox-sensitive transcription factors suchas the nuclear factor -kB and the activating protein 1. Inaddition, ROS stimulate PDE activation leading to Cyclooxygenasea decrease incytosolic cAMP and an increase in Ca2? signaling p53 Signaling Pathway in inflammatorycells, Cyclooxygenasee.g. in macrophages. Since Ca2? involving pathways mediatethe production of several pro-inflammatory substances includingTNFa, PDE inhibitors may prevent the oxidant-induced increase inCa2? levels in the macrophages, and thereby inhibit the productionof TNFa, a key protein in the regulation of inflammation . In our previous experiments, Chemical library instillation significantlyincreased several markers of oxidation of lipids andproteins in the lung homogenateof Chemical library-instilled rabbits compared to saline-instilled controls. The administration of non-selective PDE inhibitor aminophyllinedecreased the concentrations of the mentioned p53 Signaling Pathway oxidationmarkers in the lungs and improved lung functions, as well .In this study, oxidation products Cyclooxygenasewere evaluated in the lungmitochondria since they are the site of the pronounced oxidationprocesses. To evaluate the peroxidation of membrane lipids, p53 Signaling Pathway theproduction of conjugated dienes and malonyldialdehyde, an endproduct of lipoperoxidation , was determined.CD are products of the interaction of ROS with

unsaturated bonds oflipids. Thus, a significant increase Cyclooxygenaseof CD concentration p53 Signaling Pathway in the lungmitochondria isproof that LPO products react with amino acid residues and deterioratethe function of enzymes and transport systems that mayfinally lead to a loss of their integrity Cyclooxygenase. FreeMDA may react withfree thiol groups of cysteine or amino groups oflysine p53 Signaling Pathway and generate complexes reacting with thiobarbituric acid, creating thiobarbituric acid-reactive substances . A significant increase in TBARS in this study shows a higherproduction of ROS after Chemical library instillation. After olprinone treatment,COX activity increased, but was irreversible Cyclooxygenasecompared to salineinstilledcontrols.