The Essential Role Of Eotaxin And CCR3 In Airways Inflammation Of Allergic Asthma

Objective Asthma is a complex chronic inflammatory disease of the airways that involves the activation of many inflammatory and structure cells, all of which release inflammatory mediators that result in the typical pathophysiotogical changes of asthma. Such as many inflammatory cells are recruited to asthmatic airway or activated in situ there include mast cells,basosophils,eosinophils,lymphocytes which become the major source of inflammatory mediators in the airway. There have been major advances in understanding the pathway involve in the inflammatory mediators and mediator receptors. The PAF receptor was firstly cloned inflammatory mediate inflammatory receptor in 1991 and many inflammatory mediate receptors have been sequenced since then. Chemokines are very important inflammatory factors, which are attracted lymphocytes~ eosinophils~ mononuclear to the inflammatory reactionary area, then activated and realized inflammatory factors. There is little knowledge about the effecting of Eotaxin on bronchial epithelial and THland TH2 cells. Our purposes are to explore the effects of airway allergic inflammation (AAI) and eosinophil in bronchial asthma; To explore the effects of Eotaxin and CCR3 receptor by observing expression of Eotaxin and CCR3 mRNA in lung tissue of asthmatic guinea pig, and to study the mechanism of Lotaxin selectively regulating eosinophil; To observe 5-HT2AR,SPR expression in the rats lungs after intravenous IL-I f3; To investigate the changes of T helper cell (TH) subsets and its relationship to EOaxin, serum total IgE~ eosinophil counts and ventilatory function in asthmatic patients. -6- Methods I .The ovalhumin (OVA) sensitized and challenged guinea pig model of asthma was adopted ; 2. Bronchial alveolar lavage fluid (BALF) of guinea pigs was collected, in which the cells were classified and counted: 3. The pathologic samples of lung tissue were stained with HE and were processed for electron microscopy; 4.The expression of ECP in asthmatic airway mucosa was observed by immunohistochemistry (IHC); 5.After injecting IL-I f3 intravenous, then to detect the structure changes of lungs and distribution of 5-I-IT2AR and SPR in the rats; 6.The expression of Eotaxin and CCR3 mRNA in guinea lung tissue was detected by in situ hybridization ( ISH), Northern blot and RNA dot blot; 7.The CCR3 gene was detected by reverse transcription polymerase chain reaction( RT-PCR )and the product was cloned and analyzed its sequence; 8.The changes of IFN- i? IL-4. Eotaxin in peripheral T lymphocyte of asthmatic patients were observed by flow cytometry (FCM) and relationship with serum total IgE, eosinophil count and ventilatory function. Results I .ln the airways of allergic inflammation, the infiltration of eosinophils are the predominant pathologic changes of asthma; 2.The counts of eosinophils enhanced prominently in BALF of guinea pigs: 3. Electron microscopy shows the epitheliums of bronchial ,mucosa and alveolus were injured severely and the counts of eosinophils enhanced in pulmonary interstitium , meanwhile eosinophils degranulatd and the charcot-Ieyden crystals were found; 4.The expression of ECP enhanced and the activated eosinophils increased in lung tissue of asthmatic guinea pig; 5. Eotaxin and CCR3 mRNA were expressed at high levers in lung tissue of asthmatic guinea pig, which positively corrected with the expression of ECP; 6.The 5-HT2AR,SPR were highly e...