| Objective:To identify and characterize subjects with allergic bronchial asthma treated byAnti-Tumour Necrosis Factor-α (TNF-α) and Interleukin-1β (IL-1β) immunoglobulinY antibodies to understand pathophysiologic mechanisms.Methods:1. The guinea pigs were randomly divided into5groups: normal group (groupC), allergic bronchial asthma group (group M),0.1%Anti-TNF-α and IL-1β immu-noglobulin Y antibodies treatment group (group Z1),1.0%Anti-TNF-α and IL-1βimmunoglobulin Y antibodies treatment group (group Z2), budesonide treatmentgroup (group Z3).2. The guinea pig allergic bronchial asthma model was established by nebuliza-tion inhalation of ovalbuamin, followed with the treatment with inhaled Anti-TNF-αand IL-1β IgY antibodies or budesonide.3. The following allergic activities changes, such as moving, diet, cough,touching nose, sneezing, cough, respiratory rate and so on, were observed pro-andpost-treatment.4. The animals were killed at2h,4h,8h and24h after the last treatment. Thebilateral lungs were extirpated. The right lung was cut into slices and HE stained toobserve the pathologic character5. The bronchoalveolar lavage fluid (BALF) in left lung was collected andcentrifuged. The cytokines (e.g. TNF-α,IL-1,IL-4,IL-6,IL-8,IL-10,IL-13,IL-16and TGF-β) and IgE in supernatant were detected by the antibody-sandwichenzyme-linked immunosorbent assay (ELISA) method.6. All data was analyzed with SPSS statistic19.0software. Quantitative datawere recorded as mean±standard (X±SD). Differences among several groups wereconducted with one-way ANOVA. Results were considered significant when P<0.05. Results:1. It was success in construction of the guinea pig allergic bronchial asthmamodel inhaled with ovalbumin.2. Behavior observation: The animals in group M were excited, restlessness,touching the nose, sneezing and cough after inhaled the OVA. The symptom of theanimals in group Z1, group Z2and group Z3were improved after treatment.3. Pathologic changes in lung tissues:Group C: It was observed under light microscope that the structure of theairway wall and alveolar wall were integrity, and the bronchi ducts were smooth. Thealveolar cavity were clean and the epithelial cell array orderly. The capillary, thealveolar epithelial cells and other tissues in pulmonary interstitial constitute integratealveolar respiratory membrane. Infiltration of the inflammatory cells was seldomobserved around the different levels of bronchus.Group M: It was observed under light microscope that the structure of thebronchial wall, alveolar wall and alveolar cavity were damaged, with dilated capillaryand increased inflammatory cells. The alveolar cavity was broken in large area, andthe exudate with necrosis alveolar epithelial cells was detected in it. It was evidentthat the smooth muscle cells in bronchial wall hypertrophied and proliferated, and thebasement membrane was thicken and hyalinized. And it was detected the infiltrationof the eosinophilic granulocytes, lymphocytes and neutrophilic granul-ocytes inpulmonary interstitial.Group Z1and Z2: After inhaled the anti-TNF-α and IL-1β immunoglobulin Yantibodies, the degree of damage in the structure of the bronchial wall, alveolar walland alveolar cavity was diminished. The shape of the alveolar in these two groupswas more integrate than which in group M. Only a little of inflammatory cells wereobserved in alveolar cavity. The quantity of inflammatory cells in capillary wasdecreased, compared with group M. The obstruction of melicera mucus in bronchialand alveolar cavity was alleviated. It was obvious that the inflammatory cells lessinfiltrated in the bronchial cavity and pulmonary interstitial.Group Z3: The degree of damage in the structure of the lung tissues wasdiminished after inhaled budesonide. There were no obvious differences among the group Z1, Z2and Z3in pathologic changes.4. Detection of cytokine in BALF:The cytokine in BALF compared with group M: IL-1in group Z1, Z2, Z3,TNF-α in group Z1, Z2, IL-16in group Z2, Z3, IgE in group Z1, were decreased at2h after treatment (P<0.05). IL-4, IL-8and TGF-β in group Z3were decreased too(P<0.05). IL-1, IL-4, IL-6and IgE in group Z1, Z2, Z3, IL-8, IL-13and TNF-α ingroup Z2, Z3, IL-16in group Z1, Z3, were decreased at4h after treatment (P<0.05).And TGF-β in group Z1, Z2was also decreased (P<0.05). IL-1, IL-4, IL-6, IL-8,IL-13, IL-16, TNF-α and TGF-β in group Z1, Z2, Z3, IgE in group Z2and Z3, wereall decreased at8h after treatment (P<0.05). IL-1, IL-4, IL-6, IL-8, IL-13, IL-16,TNF-α, TGF-β and IgE in group C were lower than which in group M at24h aftertreatment (P<0.05). IL-1,IL-4,IL-6,IL-13,IL-16,TNF-α,TGF-β and IgE in groupZ3, IL-4,IL-6,IL-8,IL-13,IL-16,TGF-β and IgE in group Z2, IL-4and TGF-βin group Z1, were decreased at24h after treatment (P<0.05).The cytokine in BALF compared among group Z1, Z2, and Z3: There was nosignificant difference in IL-1, IL-6among the three groups2h after treatment(P<0.05). IL-4and IL-13in group Z3were lower than which in group Z1, Z2(P<0.05). IL-16and IgE in group Z2were higher than which in group Z1, Z3(P<0.05). TGF-β in group Z1was lower than which in group Z2, Z3(P<0.05). Therewas no significant difference in IL-1, IL-13and IgE among the three groups4h aftertreatment (P<0.05). IL-4and IL-6in group Z2were higher than which in group Z1(P<0.05), and IL-8, TNF-α were higher than which in group Z2, Z3(P<0.05). TGF-βin group Z2was lower than which in group Z1, Z3(P<0.05). TNF-α in group Z3waslower than which in group Z1and Z2(P<0.05). There was no significant difference inIL-1, IL-6, IL-8, IL-13and IL-16among the three groups8h after treatment (P<0.05).IL-4in group Z3was lower than which in group Z1(P<0.05). TNF-α in group Z3was lower than which in group Z1(P<0.05) and higher than which in group C and Z2(P<0.05). TNF-α and IgE in group Z2was lower than which in group Z1(P<0.05).Compared with the cytokines in group Z3at24h after treatment, IL-1in groupZ1andZ2, IL-6, IL-13and IgE in group Z1, were more higer (P<0.05), and TNF-α in groupZ1and Z2, IL-4in group Z2, were more lower (P<0.05). IL-4, IL-6, IL-13, IL-16and IgE in group Z2were lower than which in group Z1(P<0.05). IL-1in group Z1andZ2, IL-4in group Z2, IL-13and IL-16in group Z1, TNF-α in group Z2and Z3, IgEin group Z1, Z2and Z3, were higher than which in group C at24h after treatment(P<0.05)..There was no significant difference in IL-10among each group at all the timepoints. The ratio of IL-4/IL-10(0.86,1.08,0.86and0.83) in group M at2h,4h,8hand24h after treatment were more higher than which in group C (0.54,0.54,0.48,0.73) respectively. The ratio of IL-4/IL-10in group Z1(1.34) and Z2(0.98) werehigher than which in group M (0.86) at2h after treatment (P<0.05). But the ratio ofIL-4/IL-10in group Z1(0.56,0.71,0.75) and Z2(0.68,0.53,0.57) were lower thanwhich in group M respectively (P<0.05). The ratio of IL-4/IL-10in group Z3(0.74,0.67,0.57) were lower than which in group M (0.83) at2h,4h and8h respectively(P<0.05), nevertheless the ratio of IL-4/IL-10in group Z3(0.93) was higher thanwhich in group M at24h (P<0.05). In a word, the ratio of IL-4/IL-10in group Z1, Z2and Z3were lower than which in group M, and little higher than which in gruoup C,which indicated that the relative increases of IL-10in group Z1, Z2and Z3followedwith the inhibition of the allergic inflammation.5. The correlation of the cytokines in BALF:The content of TNF-α had significantly positive correlation with the content ofIL-1, IL-4, IL-8, IL-13, IL-16and IgE in BALF (P<0.05). The content of IL-1hadsignificantly positive correlation with the content of IL-4, IL-6, IL-8, IL-13, IL-16,TNF-α, TGF-β and IgE in BALF (P<0.05). The content of IL-4had significantlypositive correlation with the content of IL-1, IL-6, IL-8, IL-13, IL-16, TNF-α, TGF-βand IgE in BALF (P<0.05). The content of IL-6had significantly positive correlationwith the content of IL-1, IL-4, IL-8, IL-13, IL-16, TGF-β and IgE in BALF (P<0.05).The content of IL-8had significantly positive correlation with the content of IL-1,IL-4, IL-6, IL-13, IL-16, TNF-α, TGF-β and IgE in BALF (P<0.05). The content ofIL-10had no correlation with any other cytokines in BALF. The content of IL-13hadsignificantly positive correlation with the content of IL-1, IL-4, IL-6, IL-8, IL-16,TNF-α, TGF-β and IgE in BALF (P<0.05). The content of IL-16had significantlypositive correlation with the content of IL-1, IL-4, IL-6, IL-8, IL-13, TNF-α, TGF-β and IgE in BALF (P<0.05). The content of IgE had significantly positive correlationwith the content of IL-1, IL-4, IL-6, IL-8, IL-13, IL-16, TNF-α and TGF-β in BALF(P<0.05). The content of TGF-β had significantly positive correlation with the contentof IL-1, IL-4, IL-6, IL-8, IL-13, IL-16and IgE in BALF (P<0.05).Conclusions:1. The changes of IgE and the cytokines (e.g. IL-4, IL-6, IL-8, IL-13, IL-16,TNF-α, TGF-β) have correlation with which of TNF-α and IL-1in BALF. It has beenindicated that TNF-α and IL-1β play an important role in allergic bronchial asthma.TNF-α and IL-1β may be the initiation factor, promoter and the node in inflammatorycascade reaction in pathological process of allergic bronchial asthma.2. Anti-TNF-α and IL-1β IgY antibodies nebulization inhalation treatment couldinhibit the expression of TNF-α and IL-1β in BALF, and reduce the secretion ofinflammatory factor, such as IL-4, IL-6, IL-8, IL-13, IL-16, TGF-β, and so on. Anti-TNF-α and IL-1β Ig Y antibodies treatment could significantly attenuate the pathol-ogic damage and alleviate the symptoms in allergic bronchial asthma guinea pigmodel. Compared with budesonide treatment, anti-TNF-α and IL-1β Ig Y antibodiestreatment inhibit TNF-α and IL-1β directly and permanent for a more long time.3. Being a small sample experiment, the statistical significance in the changes ofa portion of inflammatory factor has not been detected. Furthermore, a large sampleand more perfect design should be adopted in the further study. However, Anti-TNF-αand IL-1β Ig Y antibodies treatment significantly attenuated the pathologic damage inallergic bronchial asthma in this experiment. Therefore, Anti-TNF-α and IL-1β Ig Yantibodies would be promising in the treatment with allergic bronchial asthma.4. The IL-1concentration was positive relation with IL-4, IL-6, IL-8, IL-13,IL-16, TNF-α, TGF-β, and TNF-α concentration was positive relation with IL-1, IL-4,IL-8, IL-13, IL-16, IgE in BALF. |
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