Effects Of Icarin On Mast Cell Activation And Inflammation In Rats Model With Tracheotomy Intubation By One-time Epidural Catheter

Objective(1)To observe the deregulation of mast cell in the lung of rat model with tracheotomy intubation by one-time epidural catheter,and the relationship between beta tryptase,histamine,IL-6,TNF-?and lung inflammation.(2)To observe that if the Icariin can reduce lung inflammation,inhibit the degranulation of mast cells and control the release of beta tryptase,histamine,IL-6 and TNF-?.Methods(1)54 adult male SD rats were randomly divided into 3 groups with 18 in each group: normal control group(group A),tracheotomy intubation by one-time epidural catheter model group(group B),and the icarin invention group(group C),and observe the performance of each rat.(2)To create the rat tracheotomy intubation model.(3)After 6h,the rats in group C were given intragastric administration of icariin.The group A and group B were with the same amount of 0.9% sodium chloride to instead.(4)Observeing the changes of the lung tissues in rats after the success of the model of preparation.And then,Bronchoalveolar lavage and lung tissues around the bronchial were collected with in 24 h,72h and 168 h respectively,6 mice in each time.And using the transmission electron microscope to observe mast cell ultrastructure of the rat tissue.Histamine,TNF-a and IL-6 were measured by ELISA.The lung tissues tryptase expression were measured by Immunohistochemical method.Results(1)Creation of tracheotomy intubation model in rat:With the increasing of excitation frequency,the group B appeared the performance,such as iritable,scratching their head and epidural catheter,weight growth to slow down.Pathological examination showed that the lung inflammation,the degranulation of mast cells under the electron microscope is increased obviously.Successfully established tracheotomy intubation model in rat.(2)The pathological changes of lung tissue in each group under light microscope: The lung tissue pathological of group A had no changes.The group B was damaged in varying degrees,and the damage was aggravated with the prolongation of time.At the 24 h,72h,168 h these three times in group C were lighter than those in the model group,and there was no significant difference between the three times.The Smith scores of B were higher than group A(P <0.05),and group C also more than group A;The group B and group C had no significant difference at 24h(P>0.05),but at 72 h,168h were obviously than group C(P < 0.05).(3)The degranulation of each group under electron microscope: The results show that the number of mast cells in group A is very little,and there is almost no degranulation phenomenon.In the group B,mast cells degranulation,cytoplasmic granules reduced electron density,formation of vacuoles at 24 h,72h,168 h these three times,of which is more obvious than group C.(4)Each group of?-tryptase expression: B,C group of tryptase expression in the three periods were significantly higher than group A(P < 0.05);At the 24 h group B and C there was no difference(P>0.05),however,it was significantly lower in group C than group B at 72 hours and 168 hours,especially at 168h(P <0.05).(5)The bronchoalveolar lavage fluid of histamine content: At 24 h,72h,168 h,the group B and C was significantly higher than A group(P < 0.05);the histamine content of group B had no obvious change,between B and C the two groups had no difference(P > 0.05).(6)The levels of IL-6 in bronchoalveolar lavage fluid:Both of B and C groups were higher than the normal group in 3 times(P < 0.05);Group C were significantly lower than group B.And among the three times of group B(P <0.05),the highest is at 24 h.and then decreased from 72 h to 168 h gradually,the same as group C(P < 0.05).(7)The levels of TNF-? in bronchoalveolar lavage fluid: The levels of TNF-? in lung tissue of group B is obviously higher than group A in three times(P < 0.05).Group C was significantly lower than group B(P < 0.05).And among the three times of group B,the highest is at 24 h.and then decreased from72 h to 168 h gradually,the same as group C(P < 0.05).(8)According to the results of correlation analysis between variables:Smith score and beta tryptase,TNF-alpha and IL-6 three are related,and the beta tryptase positive correlation(P < 0.05),and negatively correlated with TNF-alpha and IL-6(P < 0.05);beta tryptase and TNF-alpha and IL-6 were negative correlation(P < 0.05).Conclusion(1)In the early stage after tracheotomy,the inflammation of the lungs was observed.,then the activity of mast cells also changed,for example,the phenomenon of degranulation of mast cells is moreobvious.(2)Icariin can improve the symptoms of asthma in rat,relieve the pulmonary inflammation,inhibit its degranulation and reduce the release of beta tryptase,IL-6 and TNF-?.