| Objective: The combustion of fine particles generated by the observation and measurement of soot changes in asthma mice model by ovalbumin was established the inflammatory injury and pathological,interleukin mouse asthma model in vivo and natural immune relationship between soot PM2.5 establishment of OVA in close affect the expression of inflammatory cytokines,and to provide evidence for the way to reveal the coal PM2.5 and exacerbation of asthma and the mechanism involved.Methods: 1.PM2.5 collection: Taking bulk bituminous coal as sample,equipped with air sampler,equipped with PM2.5 filter membrane.PM2.5 was collected during coal combustion.After collection,the filter membrane was washed by ultrasonic water and filtered low temperature and vacuum dry.2.The establishment of animal grouping and model: 40 6-week male Kunming mice were randomly divided into four groups(Control group,OVA group,PM2.5 group,OVA+PM2.5 group).In sensitization stage,OVA group and OVA+PM2.5 group were intraperitoneally injected with 10 gOVA and 1mgAl(OH)3 suspension.Control group and PM2.5 group were injected equal volume of PBS for 3 times/ 1 week.The exciting stage OVA group and OVA+PM2.5 group by PBS inhalation 10mg/mlOVA configured to stimulate,Control group and PM2.5 group by PBS inhalation,30 min each time,1 time /3day,a total of six times,the PM2.5 group and OVA+PM2.5 inhalation before nasal instillation of 4?g/ ?l PM2.5 suspension 5 ?l or so each nostril.3.Tissue treatment and factor test: Tissue collection was carried out within 24 hours after the completion of the infection.Mouse bronchoalveolar lavage fluid and right upper lung tissue were collected.After centrifugation,BALF was separated,precipitated and smeared.The distribution of inflammatory cells was observed under microscope.Supernatant,enzyme-linked immunosorbent assay(ELISA)was used to detect IL-4,IL-7,IL-8,IL-17,TGF-beta,IFN-gamma,serum specific IgE and IgG1.The level of IL-7mRNA and IFNgamma mRNA in lung tissue was detected by RT-PCR.Results: 1.In Control group: normal behavior,no significant changes were observed.In OVA group: caught the ear of facial behavior,breathing with dramatic ups and downs.In PM2.5 group: occasionally catch ear,irritability.In OVA+PM2.5 group: mice breath can smell and deepen quickly,most caught the ear of behavior,dysphoria and irritability.2.The distribution of inflammatory cells in mice in BALF,OVA group,Lym,Neu,Mon,Eos cell count increased significantly compared with Control group(p<0.05);PM2.5 group Mon,Lym,Neu,Eos,the total number of cells was higher than Control group(p<0.05);OVA+PM2.5 group Mon,Lym,Neu,Eos,cell number compared with the other three groups were significantly increased(p<0.05).Control group lung tissue sections were stained microscopically had no obvious change;OVA group showed a large number of eosinophils gathered around the bronchial wall and visible mononuclear cells and lymphocytes,epithelial cell deformation,shedding;PM2.5 group appeared in alveolar eosinophils accumulation,visible alveolar hyperemia,compared with the OVA group,less inflammatory cells;OVA+PM2.5 group gathered a large number of inflammatory cells in lung tissues,around the bronchi,interstitial infiltration,bronchial epithelial exfoliation,exudation tube.Electron microscope results: Control group of capillary lumen visible part of neutrophils,type II epithelial cell cytoplasm showed rich osmiophilic lamellar bodies;OVA group individual capillary lumen neutrophils,II cells in a small amount of osmiophilic lamellar bodies,small lamellar body evacuation.The PM2.5 group mitochondria had no obvious abnormalities,cell surface microvilli decreased and disappeared,showed visible macrophage infiltration;type I epithelial cells of individual swelling significantly,type II epithelial cells in the cytoplasm of a small amount of osmiophilic lamellar bodies,lamellar body evacuation,mitochondrial swelling,disappearance of cell surface microvilli observed.In OVA+PM2.5 group,I type individual epithelial cell swelling significantly,type II epithelial cells in the cytoplasm of a small amount of osmiophilic lamellar bodies,lamellar body evacuation,mitochondrial swelling,occasional macrophages,alveolar septa obviously individual collagen fiber bundle.The results of Elisa,IL-4:Control were lower than the other three groups(p<0.05),OVA+PM2.5 group was higher than that of other three groups(p<0.05,F=26.884);IL-7:Control group and PM2.5 group had no significant difference(p=0.776),the other group arranged from low to high as PM2.5<OVA<OVA+PM2.5(p<0.05),F= 15.327;IL-8:OVA+PM2.5 group was higher than that of Control group,OVA group and PM2.5 group(p<0.05),Control group was lower than that of OVA group and PM2.5 group(p<0.05,F=45.324);group IL-17: four from low to high order as follows: Control<OVA<PM2.5<OVA+PM2.5(p<0.05),F=26.744;IFN-: Control gamma group and PM2.5 group had no significant difference(p=0.071),the other groups were different(p<0.05,F=17.526);TGF-beta: Control group and PM2.5 group had no significant difference(p=0.144),the other groups were different(p<0.05,F=144.332).IgE: the four groups were arranged from low to high as follows: Control<OVA<PM2.5<OVA+PM2.5,(p<0.05),F=19.374;OVA-IgG1:Control group were lower than those of other groups(p<0.05),OVA group was higher than PM2.5 group(p<0.05),F=33.611.RT-PCR detection,IL7 mRNA group only between the upward trend(p>0.05),F=0.028,IFN-gamma mRNA Control group is higher than the rest of the other groups,(p<0.05),F=63.651.Conclusion: Coal burning PM2.5 can change Treg/Th balance in OVA induced asthma mouse model by influencing IL17 related inflammatory factors,and then aggravating respiratory tract inflammatory injury and tissue structure change in asthmatic mice. |
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