| Objective:To investigate the mechanism of Anti-Tumour Necrosis Factor Alpha and Interleukin-lbeta Immunoglobulin Y Antibodies nebulization inhalation in local tissue surface in treatment to allergic bronchial asthma.Methods:Ovalbuamin nebulization inhalation was applied to establish stable guinea pig allergic bronchial asthma model.The animals were randomly divided into 4 groups:normal group(group A),allergic bronchial asthma group(group B),0.1% Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta Immunoglobulin Y Antibodies treatment group(group C),1.0%Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta lmmunoglobulin Y Antibodies treatment group(group D).The following allergic activities changes of the animals were observed by humanbeing:dieting,moving,scrashing nose,cough,sneezing and so on.The animals were killed after treatment at different time points:2h,4h,8h,24h and the color shape as well as some other indexes of the lung tissue were observed.The apex of lung,the main body of lung and the base of lung were collected to make pathological sections.And the pathological sections were HE stained to study the situation of the following indexes:epithelial cells of pulmonary alveoli,the structure of pulmonary alveoli wall.The density,shape,blocking situation and edema in mucous membrane of capillary were also studied.The kinds and accounting of inflammatory cells infiltrating aroud the bronchus were studied.The situation of content in pulmonary alveoli was observed too.The bronchovesicular in left lungs were lavaged by isotonic Na chloride,the bronchoalveolar lavage fluid(BALF)were collected and centrifugated to get the deposied cells.The deposied cells were made into cells smears and were Wright's stained to account eosinophilic granulocytes,neutrophilic granulocytes,lymphocytes,macrophages and so on.The total protein amount in supernatant of BALF was measured by Lowrys method.The antibody-sandwich enzyme-linked immunosorbent assay(ELISA)method was used to detecte the amount ofTNF-α,IL-1,IL-4,IL-8,IL-10,IL-13 in supernatantofBALF.Collect the datas of cytokine/total protein ratio in different groups at various time points. Results:Activites changes:The behavior,moving,dieting and so on in Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta Immunoglobulin Y Antibodies treatment group was turning to normal after being treated 3 days.Pathophysiology changes in lung tissues:,Group A:When be observed under light microscope the structure of alveolar ducts and alveolar wall were integrate,the alveolar space were clean and hyaline.The alveolar epithelial cell array orderly.And the alveolar epithelial cell,capillary and other interstitial form integrate alveolar respiratory membrane.The density of capillaries in alveolar wall were intensive and the alveolar ducts were smooth.Regular shape erythrocyte can be seen in it.Group B: When be observed under light microscope the structure of alveolar ducts and alveolar wall were damaged.The alveolar space were full of transudate,lots of caducous necrosis alveolar epithelial cells and oozy leucocyte can be seen in all the sight field. The alveolar wall was thickened,the capillary in alveolar wall were distorted or dilated and congested accompanied with leucocyte impaction,erythrocyte siltation and useful capillaries decrease.The above were caused by lung interstitial edema and inflammatory cells infiltration.Group C and D:After different concentration treatment of Anti-Tumour Necrosis Factor Alpha and Interleukin-lbeta Immunoglobulin Y Antibodies,the degree of damage in alveolar ducts and alveolar wall were slighter than that in group B.little inflammatory cells can be seen floating in alveolar space.Melicera mucus plug in bronchial lumen and pulmonary alveoli were significantly less than that in group B.The Bronchial lumen in group C and D were more regulate and Inflammatory cells infiltration can seldom be seen aroud it.Recovery of tunica mucosa bronchiorum epithelium was obvious.There is no significant difference between group C and D.Cells content Change in BALF:The number of eosinophile granulocyte, neutrophilic granulocyte,lymphocyte,in group C are significantly decreasd (2h,4h,8h,P＜0.05).While the number of macrophage is significantly increased (2h,4h,8h,P＜0.05)compared with group B.The number of eosinophile granulocyte in group D is significantly decreased than that in group B(2h,4h,8h,P＜0.05).The number of neutrophilic granulocyte in group D is significantly decreased than that in group B(2h,8h P＜0.05)The number of lymphocyte in group D is significantly decreased than that in group B(2h,8h,24h,P＜0.05)While the number of macrophage is significantly increased(2h,8h,P＜0.05)The change of cytokine in BALF:Compared with group B the cytokine(pg) of protein(μg)in BALF:IL-1 in group C and group D was significantly decreased after beingtreated2hlater(group B:0.040±0.014,group C:0.013±0.001,group D:0.019±0.006).IL-8 was significantly decreased too(group B:0.162±0.192,group C:0.015±0.001,group D:0.025±0.007).IL-4 was significantly decreased only in group C(group B:0.104±0.011,group C:0.052±0.036).Compared with group B IL-1 in group C was decreased(group B:0.023±0.007,group C:0.015±0.002) afterbeing treated 4h later.IL-4 was decreased(group B:0.094±0.046,group C: 0.076±0.055).IL-8 was decreased(group B:0.028±0.009,group C: 0.017±0.003).IL-13 was decreased(group B.0.030±0.010,group C:0.018±0.009). TNF-αwas decreased(group B:0.072±0.012,group C:0.063±0.035).While IL-10 was significantly decreased than that in group D(group B:0.242±0.102,group D: 0.337±0.103).Compared with group B IL-10 in group C was significantly increased (group B:0.402±0.098,group C:2.846±0.445,P＜0.05)after being treated 8h later. IL-10 was increased too in group D(group D:0.497±0.289).TNF-αwas decreased in group D(group B:0.100±0.019,group D:0.084±0.014).After being treated 24h later compared with group B IL-1 in group C was significantly decreased(group B: 0.028±0.012,group C:0.011±0.005).IL-4 was significantly decreased(group B: 0.181±0.086,group C:0.070±0.010).IL-8 was decreased(group B:0.027±0.005, group C:0.018±0.006).IL-13 was significantly decreased(group B:0.034±0.010, group C:0.017±0.004).TNF-αwas decreased(group B:0.102±0.034,group C: 0.077±0.029).IL-10 in group D was significantly increased(group B:0.365±0.247, group D:0.765±0.264).TNF-αwas decreased(group B:0.102±0.034,group D:0.088±0.020).After being treated 8h later the IL-10/IL-1 in group C and group B (42.75:19.14).The IL-10/IL-4 in group C and group B(5.16:2.73).The IL-10/IL-8 in group C and group B(36.49:16.08).The IL-10/IL-13 in group C and group B(23.92:10.58).The IL-10/TNF-αin group C and group B(4.87:4.02) significantly or obviously increased.At the same time points the IL-10/TNF-αin group D and group B increased slightly(5.92:4.02).After being treated 24h later the IL-10/IL-1 in group C and group B(25.64:13.04).The IL-10/IL-4 in group C and group B(4.03:2.02).The IL-10/IL-8 in group C and group B(15.67:13.52).The IL-10/IL-13 in group C and group B(16.59:10.74)significantly or obviously increased.After being treated 24h later the IL-10/IL-1 in group D and group B(14.17: 13.04).The IL-10/IL-4 in group D and group B(3.12:2.02).The IL-10/IL-8 in group D and group B(15.30:13.52).The IL-10/IL-13 in group D and group B(21.25: 10.74).The IL-10/TNF-αin group D and group B(8.69:3.58)significantly or obviously increased.Conclusions:In this experiment the following items were explored:The integrate activites behavior of all animals in each group,pathophysiology changes in lung tissues;the amount changes of inflammatory cells(eosinophilic granulocyte, neutrophilic granulocyte and lymphocyte),inflammatory cytokines(IL-1β,TNF-α,IL-4, IL-8,IL-13)and anti- inflammatory cytokines(IL-10)in BALF.The effect of proinflammatory cytokines in allergic bronchial asthma,the relationship between proinflammatory cytokines and other cytokines in allergic bronchial asthma,the mechanism of different concentration Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta Immunoglobulin Y Antibodies nebulization inhalation in treatment to allergic bronchial asthma guinea pig model were discussed.And it has been primarily proved that TNF-a and IL-1βplay an important part in allergic bronchial asthma.They could be promoter,acceleration factor,enhanser as well as tissue injury factor in the inflammatory cytokines net.Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta Immunoglobulin Y Antibodies nebulization inhalation treatment could significantly attenuate the symptoms and the inflammatory pathology in allergic bronchial asthma guinea pig model.Meanwhile the amount of eosinophilic granulocyte,neutrophilic granulocyte and lymphocyte in BALF were significantly or obviously decreased.The content of IL-1β,TNF-α,IL-4,IL-8,IL-13 were decreased too.The content of IL-10 and IL-10/IL-1β,IL-10/ TNF-α,IL-10/ IL-4,IL-10/ IL-8,IL-10/ IL-13 were increased.By changing the amount or the ratio of the inflammatory cytokines above,the symptoms and signs of allergic bronchial asthma were decreased.Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta lmmunoglobulin Y Antibodies nebulization inhalation treatment was applied in this experiment to treat allergic bronchial asthma guinea pig model.And the results indicates that Anti-Tumour Necrosis Factor Alpha and Interleukin -lbeta Immunoglobulin Y Antibodies local passive immunity treatment to allergic inflammation is of bright future.
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