Research on the Efficacy of Aminophylline on Phagocytosis of Apoptotic Eosinophils by A549 Cells
|School||Anhui Medical University,|
|Keywords||Aminophylline A549 cells Eosinophils IL-6 IL-8|
Background eosinophil infiltration is important manifestation of bronchial asthma pathogenesis, the eosinophil outcome affect the prognosis of asthma. Cell death pathways, there are two: First, death, another apoptosis. Proinflammatory substances which apoptosis can make because of the integrity of the cell membrane is not compromised. Apoptosis, phagocytosis because expression of a series of proteins in the cell membrane that can be swallowed. Eosinophil apoptosis with phagocytosis airway epithelial phagocytosis can reduce the inflammatory response of the airway in bronchial asthma. Aminophylline is an effective drug for the treatment of bronchial asthma, recent experimental studies have shown that in addition to a role of bronchial relaxation, aminophylline, immune and anti-inflammatory role. Already reported in the literature dexamethasone can upregulate epithelial cell phagocytosis of apoptotic eosinophils capacity, and there is a concentration-dependent effect. With effective drug for the treatment of bronchial asthma, for the aminophylline whether intervention in the epithelial cells, thereby affecting the ability of phagocytosis of apoptotic eosinophils unclear. Reported aminophylline can inhibit the secretion of inflammatory cytokines, aminophylline phagocytosis of apoptotic epithelial cells eosinophils intervention role has not been reported whether inhibition of the secretion of inflammatory factors related. Therefore study the role of the eosinophils intervention aminophylline phagocytosis of apoptotic epithelial cells and IL-6, IL-8 secretion, research aminophylline role in the treatment of bronchial asthma and anti-inflammatory the role of the new ideas of great significance, and provide a new basis for bronchial asthma clinical treatment. Purpose build a model of the A549 cells phagocytosis of apoptotic eosinophils, epithelial cells engulf apoptotic eosinophils intervention cell model for further study of aminophylline. A549 cells phagocytosis of apoptotic eosinophils model based intervention research aminophylline A549 cells phagocytosis. And about whether the concentration-dependent and time-dependent relationship. Research aminophylline A549 cells were stimulated by lipopolysaccharide and apoptotic eosinophils secrete inflammatory factors whether the inhibition of the existence of concentration dependence. 4, the role of eosinophils intervention aminophylline phagocytosis of apoptotic epithelial cells and inhibit the secretion of inflammatory factors are related. Continued passage of A549 cells by culture, different concentrations of aminophylline (0,10-3,10-4,10-5 mol / L) was added to A549 cells 24, 48 and 72 hours prior to phagocytosis experiments; magnetic cell separation column (MACS Column) CD16 Beads antibody, CD15 the bead antibody separation purification of human peripheral blood eosinophils in vitro culture 48 hours after natural aging by morphological and AnnexinV-FITC kit eosinophilic Identification of granulocyte apoptosis, then add to the A549 cells after aminophylline intervention, the use of o-dianisidine peroxidase staining engulf apoptotic eosinophils A549 cells, and then be with hematoxylin nuclear lining into. The use of Olympus inverted microscope and Diagnostic Instruments-Spot insigh digital image acquisition the camera system dynamically observe the the A549 cells engulf apoptotic eosinophils phenomenon, and phagocytic rate statistics on fixed and stained cells in the light microscope. Other different concentrations of aminophylline (0,10-3,10-4,10-5 mol / L), respectively, to intervene in A549 cells, A549 and apoptosis of eosinophils, A549 and lipopolysaccharide, harvested cultured on supernatant was detected by radioimmunoassay interleukin IL-6 and IL-8 concentration. Results 1 A549 cells were aminophylline intervention after 24, 48 and 72 hours, the A549 phagocytic ability decreased with time. A549 cells phagocytic capacity aminophylline 10-3 mol / L the phagocytic capacity minimum concentration intervention, 10-4 mol / L ,10-5 mol / L significantly increased. A549 cells, phagocytosis of apoptotic eosinophils compared with non-phagocytic A549 cells, IL-6, IL-8 secretion was no significant difference. A549 cells compared with lipopolysaccharide-stimulated IL-6, IL-8 secretion values ??decreased significantly. 4, aminophylline can inhibit lipopolysaccharide-stimulated A549 cells, IL-6, IL-8 secretion and concentration dependency. Conclusions, aminophylline A549 cells phagocytosis of apoptotic eosinophils, and has a time-and concentration-dependent effects. , Aminophylline through inhibition of IL-6, IL-8 ways to play the role of anti-inflammatory, and has the effect of a concentration-dependent manner. The A549 phagocytosis of apoptotic eosinophils can reduce the inflammatory response, bronchial inflammation in asthma control is of great significance.