Objective:1.Establish mice model of asthma by OVA injection and sensitization.Observed Lung histopathological changes in the mice with the bronchial asthma in theacute, chronic mice of asthma and the normal mice.2.Observed the changes ofPPAR-γ,EOS,IL-13and VEGF level in the mice with the bronchial asthma in acute,chronic, remission period and the normal mice.3. Explore the role of PPAR-γ in theairway inflammation and airway remodeling of bronchial asthma.Methods:30mice which4~6weeks old,18~22g health, female, SPF, wererandomly divide into acute asthma group (acute group,10mice), chronic asthmagroup(chronic group,10mice), and the normal control group(control group,10mice), toestablish asthma model with the method of egg albumin(OVA) sensitization andstimulate: acute, chronic group in the first0,7,14days with20μgOVA+150μlAI(OH)+350μlNS(100mg/L) intraperitoneal injection and sensitization;27,28,29days will be Intranasally to stimulate by OVA-NS(50μl,2g/L);Acute group weresacrificed at30days24hours; chronic group aerosol inhalation of2%OVA every dayin the31~59days; In normal group used normal saline intraperitoneal injection andaerosol inhalation, treatment time was the same to chronic group; The first60daysexecuted chronic, normal group. Observed HE staining of lung tissue pathology airwayinflammation, total trachea wall thickness (Wat/pbm), tracheal wall thickness(Wai/pbm)and airway smooth muscle thicknes(sWam/pbm)in mice; bronchoalveolarlavage fluid (BALF) cell sorting; biotin double-antibody sandwich enzymelinkedimmunosorbent assay interleukin-13(IL-13) and vascular endothelial growth factor(VEGF) levels; immunohistochemical method for the determination of the peroxisomeproliferator-activated receptor γ (peroxisome proliferators activated receptor PPAR-γ)protein expression.The obtained empirical datum uses the SPSS17.0Software to carryon the analysis. Results:1. The performance and comparison of asthma mice model: the mice of acuteandgroup both apparent restlessness, easily frightened, shortness of breath, wipe nasal,sneezing, lip tail purple cyanosis, activity frequently, fur shadowbane, abdominalmuscle spasm, incontinence, etc; chronic group showed more especially noticeable, therhythm of breathing not neat, fell motionless, limbs undermines. normal group did notappear afore-mentioned symptoms.2. Pulmonary histopathologically observation: acute and chronic group comparedwith normal group showed lung surface swelling, increase in size, cut-side visible oozywhite foam. Lung biopsy HE dyeing appear bronchial lumen narrow, mucosa edema,mucous glandular hyperplasia, mucous membrane, the alveolar every thickening, edemairregular shape, the alveolar space fusion expanded. acute Group bronchial smoothmuscle, blood vessels around the mucosa and submucosa shows inflammatory cellinfiltration; chronic group in the acute stage of the pathological basis of the gassubepithelial fibrosis, goblet cell hyperplasia, airway smooth muscle cell proliferation,hypertrophy, airway smooth muscle cell migration and vascular remodeling andangiogenesis, ie, airway remodeling. Normal group normal mouse airways and alveolarstructures and bronchial epithelial integrity, and no significant inflammatory cells andairway remodeling change.3. Airway morphology analysis: acute,chronic and normal mice of total tracheawall thickness(Wat/pbm)9.64±1.36μm~2/μm,15.25±1.43μm~2/μm,5.59±1.41μm~2/μm,chronic group’s Wat/pbm more than acute and chronic group was significantlyincreased (P <0.01) difference was statistically significant; acute group of Wat/pbmthan normal group was significantly increased (P <0.01) difference was statisticallysignificant;Tracheal wall thickness (Wai/pbm) of4.68±1.22μm~2/μm,9.49±1.19μm~2/μm,2.38±0.62μm~2/μm, acute group’s Wai/pbm more than acute and normalgroup was significantly increased (P <0.01) difference was statistically significant,acute group’s Wai/pbm more than normal group is significantly increased(P<0.01)difference is statistically significance; airway smooth muscle thickness(Wam/pbm)thickness, respectively,10.64±1.30μm~2/μm,15.67±2.02μm~2/μm,5.05±0.85μm~2/μm, chronic group’s Wam/pbm mroe than acute and normal group issignificantly increased (P<0.01) difference is statistically significant. acute group’sWam/pbm more than normal group is significantly increased (P <0.01) difference isstatistically significant. 4. Serum PPAR-γ level: Acute group mice of PPAR-γ-positive rate is11.23±3.27,chronic group is13.05±0.82, two groups of lung tissue can be seen a large number ofPPAR-γ in expression, mainly in the airway epithelium, submucosal, and around theairway and alveolar regions cells, the expression of PPAR-γ is mainly in the cytoplasm;normal group mice PPAR-γ-positive rate is7.63±0.74, less positive expression in lungtissue; chronic group higher than acute and normal group (P <0.01) in PPAR-γ level, thedifference is statistically significant; acute group higher than normal group (P<0.01) inPPAR-γlevel, the difference is statistically significant;5. BALF EOS%: acute group’s EOS%is13.69±3.37%, chronic group is23.33±2.03%,normal group is2.80±0.65%,chronic group higher than acute and normalgroup (P<0.01), the difference was statistically significant; acute group higher thannormal group (P<0.01), the difference was statistically significant;6. BALF IL-13level: acute group for77.69±25.55ng/L, chronic group for246.14±15.77ng/L, normal group for32.24±9.61ng/ml. chronic higher than acute and normalgroup (P<0.01), the difference was statistically significant; acute group higher thannormal group (P<0.01), the difference was statistically significant;7. BALF VEGF level: acute group for40.45±3.76ng/ml, chronic group for59.82±16.97ng/ml, normal group for27.75±3.78ng/ml. chronic higher than acute andnormal group (P <0.01), the difference was statistically significant; acute group higherthan normal group (P<0.01), the difference was statistically significant;8. Chronic group:BALF IL-13and VEGF level positively correlated (r=0.944,P<0.01), PPAR-γ with the total trachea thickness is positively correlation (r=0.863, P<0.01),the difference is statistically significant.Conclusion:1.The Airway remodeling mode of the mouse with Asthma was built successfully,pathological changes of acute phase and chronic stage accord with the Change ofpathophysiolog about Asthma.2.IL-13,VEGF and EOS play a role in the occurrence of Asthma, participate ininflammatory reaction of the Asthma and Airway remodeling procedure, and their levelcan reflect the Severity of the Asthma to some extent.3.PPAR-γ has a hand in the whole process of chronic inflammatory in Airwayand Airway remodeling, in connection with the Severity of the Asthma4.Expression of PPAR-γin bronchialasthma is in connection with EosinophilDependent on Th2cellã€IL-13secreted from Th2cell and the level of VEGF.,which points out that Action of PPAR-γfor the occurrence of Asthma is Multiple target.Allthis above provide the Laboratory of theoretical basis about PPAR-γactivator applied toTreatment of Asthma. |