The Studay On Expression And Significance Of Related Cytokines Of Th17/Treg Cell In Severe Pneumonia Induced By Pseudomonas Aeruginosa

Objective: Discuss different doses of pseudomonas aeruginosa to construct the meaning of severe pneumonia SD rats model Through two aspects, the alveolar flushing fluid and peripheral blood observation of TH17/Trge cells in pseudomonas aeruginosa infection of SD rats with severe changes in lung inflammation. Discuss two kinds of cells play a role in the host immune system. Methods.1) Pseudomonas aeruginosa is provided by guilin medical college affiliated hospital clinical laboratory,streak plated on blood plate. In 37 ℃ CO2 hatch training contents, Switching a cultivation every 24 hours in order to ensure the activity of pseudomonas aeruginosa. Using spectro photometric analysis and colorimetry 2 hours before. With physiological saline as 0.5* 10 ^ 10 cfu/ml, .2* 10 ^ 10 cfu/ml,2.0* 10 ^ 10 cfu/ml concentration of three kinds of mixed suspension.2)8₁0 weeks 40 SD rats, Weighting 300g+20g,Then they were divided into experimental group and control group randomly,30 rats and 10 rats respectively. Intraperitoneal injection of 10% chloral hydrate（4ml/kg）, The control group endotracheal intubation note 0.2 ml of normal saline After anesthesia The experimental group was divided into three groups, low dose group, middle dose group, high dose endotracheal give 0.2 ml mixed suspension, which Concentrations （?） To be put to death in rats after 72 hours, obtaining alveolar lavage fluid and lung tissue and peripheral blood HE dyeing observation lung tissue pathological changes after PA infection.3) 8₁0 weeks 80 SD rats Weighting 300g±20g,Then they were divided into experimental group and control group randomly,40 rats respectively.For experimental group,Tracheal push note 0.2 ml concentration is 1.2* 10 ^ 10 cfu/ml of suspension. The control group inject Sodium Chloride at the same dosage. After processing,abtaining abdominal aorta, respectively, and put to death in the rat blood, and to return alveolar lavage, in 12 h,24 h,3 d,5 d,7 d respectively. BALF detect the changes of inflammatory cells; Flow cytometry test peripheral blood of rats with Trge Th17 cell level. Results:1) Clinical symptoms and pathological lung infection:In PA bacteria inoculated in experimental rats were pneumonia clinical manifestations but its different levels except 2rats in the Low dose group.1 in midle dose group,6 in high dose group around the nose and mouth with blood, And high dose group has three die within 12 hours. The control group no pneumonia clinical manifestations, and no death.2) Three groups of rats inoculated PA bacteria occuring lobular pneumonia under light microscope. The middle and lower lobectomy visible blend of hemorrhagic lesions And there is significant difference. Peripheral blood and alveolar lavage fluid bacterial culture showed positive differences are significant. Dose group and high dose group was significantly higher in low dose group and the control group. Lobectomy in grinding per gram bacteria content of lung tissue were greater than 10 ^ 5 cfu.3)Severe pneumonia in rats model of PA infection neutrophil increased significantly after 12 h, and peaking （28.09 ±0.68* 10 A 6 on the third day; Th17 cells rise rapidly in the early stages of the infection 2.0±0.14）%.Expression trend and the expression of the ANC is correlation.Treg cells expressed in the early stages of the infection（3.21±0.10）% And then gradually incresed with the progress of the infection. Th17 cells and Treg cells in peripheral blood of change is consistent with the change of alveolar lavage fluid, And any time period and the expression of control group there were significant differences （P< 0.05）. Comparing two group at each time period also have statistically significant differences （P< 0.01） Conclusion:1) the typical experiment made clear mold infection site and pathological changes, has a good stability and repeatability; 2) in the mouse model of severe pneumonia infection in PA Thl7 cells involved in the process of severe pneumonia occurrence, development, and has a proinflammatory role; Treg cells in the process of the occurrence and development of pneumonia have anti inflammatory effect, in the late stage of infection both gradually reach dynamic equilibrium. About the delivery time is early or super early, and can prevent or reverse the medicine two kinds of cells in the expression of severe pneumonia reduce lung substantial damage remains to be further tested.