Research On The Efficacy Of Aminophylline On Phagocytosis Of Apoptotic Eosinophils By A549 Cells

BackgroundEosinophils(Eos) infiltrating is an important appearance during asthma invasion,and the turnover of Eos to affect asthma turnover.There are two pathways about cell metabolism:one is death,another is apoptosis.Beacause the cell membranes of apoptosis cells are Integrity,so the inducing inflammation materials can not reveal from cells.Then a lot of proteinums expression on the cell membranes sothat apoptosis cells can be phagocytize.that is to be say, apoptosis Eosinophils can be phagocytized by airway cellula epithelialis which can phagocytize other cells.The process may lessen inflammation of airway during asthma invasion.Aminophylline(AM) is one utility medicine for asthma, recently years,many experiments and researchs indicate that Aminophylline can possess not only relax bronchus but also immune and anti-inflammatory.Now some literatures report dexamethasone can up-regulation the capability of epithelial cell phagocytize apoptosis eosinophils,and exist concentration rely effect. Aminophylline can also effect phagocytes and eosinophils.To cure asthma the utility medicine including dexamethasone as well as aminophylline. Can aminophylline effect epithelial cells phagocytize apoptosis eosinophils ?We can not know the action and mechanism about this.Now having documents report aminophylline can inhibit many inflammatory factor secretion,but if aminophylline effect epithelial cells phagocytize apoptosis eosinophils correlate effect IL-6,IL-8,This investigation is very important because it have important significance about aminophylline incure asthma and new pathway about aminophylline anti-inflammatory,and it can be to provide new evidence for asthma clinical iatreusis.Objectives1. To construct A549 cells phagocytize apoptosis eosinophils model, For study aminophylline effect epithelial cells phagocytize apoptosis eosinophils to provide cells model.2. Basic of the model about A549 cells phagocytize apoptosis eosinophils,invest to aminophylline effect epithelial cells phagocytize apoptosis eosinophils.And understand if it have concentration relying or time relying.3. Invest whether or not aminophylline inhibit A549 cells secrete inflammatory factor which be stimulus by lipopolysaccharide(LPS) and apoptosis eosinophils. Whether or not it existence concentration relying.4. Whether or not aminophylline effect epithelial cells phagocytize apoptosis eosinophils correlate with inhibit secrete inflammatory factor.MethodsA549 cells were cultured and passsaged to last, Adds aminophylline of different concentrate on (0,10-3,10-4,10-5mol/L) to A549 cells before engulf experiment at 12,24,72h. Eosinophils were purified from normal donors by magneticactivated separation column (MACS Column),CD16 magnetic bead antibody and CD15 magnetic bead antibody.The cells were cultured for 48h and and apoptosis induced. then add the A549 cells which treated by aminophylline. Morphology and AnnexinV-FITC kit were used to accredit apoptotic eosinophils. Eosinophils stained for peroxidase with o-dianisidine and A549 cells stained with haematoxylin facilitated.Olympus inverted microscope and Diagnostic Instruments-Spot insight were used to dynamic observe the phenomenon of A549 cells engulf apoptotic eosinophils,calculate the ratio of engulfment under the light microscope with the cells which were be fixed and stained. Add aminophylline of different concentrate on (0,10-3,10-4,10-5mol/L) to A549 cells,A549 cells and lipopolysaccharide(LPS),A549 cells and apoptotic eosinophils. All these supernatant liquid were cultured for collected and using radioimmunoassay kit to detect the concentration of IL-6,IL-8.Result1. When A549 cells were effected by Aminophylline for 24,48,72 hours, these cells, engulfment decline time by time.2. Which A549 cells were effected by Aminophylline with 10-3 mol/L, these cells, engulfment were most lowness with 10-3 mol/L,and hoist with 10-4 mol/L,10-5 mol/L.3. After A549 cells engulf apoptotic eosinophils, nothing changes about IL-6,IL-8. But decline with lipopolysaccharide.4. Aminophylline can inhabit A549 cells secrete IL-6,IL-8 and in concentration manner.Conclusions1.Aminophylline inhibited the A549 cells engulf apoptotic eosinophils.2.Aminophylline can to bring into full play anti-inflammation effective by inhibited the secretion of IL-6,IL-8 .3. Which A549 cells phagocytize apoptosis eosinophils can depress inflammation,and it may be very important for control asthma.