Effects Of ICS On The Degranulation Of Mast Cell In The Airway Of Asthmatic Rats

Objective1. To observe the degranulation of mast cell in the airways of the asthmatic rats, and the relationship between airway inflammation of asthma and β- tryptase and IL-6.2. To observe that if the ICS- budesonide can reduce airway inflammation, reduce the hyperplasia of mast cells, protect the membrane of mast cells, inhibit the degranulation of mast cells, and control the release of the beta tryptase and IL- 6.3. To compare the budesonide and mast cell stabilizer- tranilast on the treatment and prevention of asthma, and on the inhibition of mast cells’ degranulation.4. To explore budesonide and tranilast whether there is synergy on the treatment and prevention of asthma.Method1. 40 adult male SD rats divided into five groups: normal control group(group A), asthma model group(group B) and budesonide intervention group(group C), tranilast intervention group(group D) and budesonide + tranilast intervention group(group E),each group of 8, and observe and record the performance of each rat.2. To create asthma model of subacute by using the ovalbumin to sensitize and inspire. The control group with 0.9% sodium chloride to instead.3. Drug intervention group respectively with budesonide to pump atomization and tranilast to gavage, The control group with 0.9% sodium chloride to dispose.4. Take the serum, then using the ELISA method to detect the serum content of beta tryptase and IL- 6. Gathering the lung tissue around the bronchial, and then using Western Blot to test the levels of beta tryptase in lung tissue of asthmatic rats. Using pathology examination with HE staining to observe the airway inflammation of each rats. A pplying toluidine blue stain to observe the change of morphology and counting of mast cell in the lung tissue between each groups.5. In the experimental process, We found that tranilast exist the possibility of damage urinary system. So when the end of the experiment we collecting kidney and bladder specimen of each rats with HE staining, and then screening for pathological detection to presence of inflammatory lesions in the kidney and bladder.Result1. Construction of subacute asthma model in rats: With the increasing of excitation frequency, the rats of asthma model group(group B) appeared the sample performance of asthma. Such as irritable, scratching their heads,shortness of breath, sneeze, weight growth to slow down. Pathological examination showed that the inflammation of airway is obviously, Eosinophilic cells is increased;The number of mast cells in lung tissue and around the airway are increased, the degranulation of mast cells under the electron microscope is increased obviously. Successfully established subacute asthma model in rats.2. Mast cell count: The mast cell count of group B is obviously higher than group A. And that indicator of group C, group D and group E are all significantly reduced. Among the three groups, the most obviously reduced is group E.But it’s still more than group A.3. The mast cell degranulation of each group under the electron microscope: The results show that the number of mast cells in group A is very little, and there is almost no obvious degranulation phenomenon. But the hyperplasia and degranulation of mast cells are evident. And those indicators of group C, group D and group E are all significantly more ease than group A.4. The levels of beta tryptase in lung tissue: The levels of beta tryptase in lung tissue of group B is obviously higher than group A. Group C, group D and group E are significantly lower than group B. And among the three groups the lowest is group E. But they are still higher than that of group A. And the differences are statistically significant(P < 0.05).There was no significant difference between group C and group D.5. Serum content of beta tryptase and IL- 6: the levels of beta tryptase and IL- 6 in serum of group B is up from group A.Group C, group D and group E are all significantly lower than group B. Among the three groups the lowest is group E. But they are still higher than those of group A. And the differences are statistically significant(P < 0.05). There was no significant difference between group C and group D. Correlation analysis showed that beta tryptase and IL- 6 are positively correlated.6. In the group with tranilast intervention( group D), we found a case of cystitis. In the experimental process, the experimental rats was found to drink more water and output more urine. Pathological results showed that the bladder mucosal of one of the experimental rats is edema, with the inflammatory cell infiltration in the bladder interstitial, and the lymphocytes is the main inflammatory cells, And it is considered interstitial inflammation on bladder.Conclusion1. ICS- budesonide can improve the symptoms of asthma in rats, relieve the airway inflammation, reduce the number of mast cells proliferation, inhibit its degranulation and reduce the release of beta tryptase and IL- 6.2. On laboratory index, budesonide with tranilast combined application is better than that of budesonide or tranilast separate applications.3. Under certain conditions, tranilast may cause bladder interstitial inflammation.