The Effect Of Budesonide And Monkelust On Expression Of Clara Cell Secretory Protein (CC16) In Asthmatic Mice

Objective:To investigate the Clara cell secretory protein expression in the blood and lung tissue of asthmatic mice and the effect of budesonide and monkelust on CC16 expression,and to analyze its role in asthma pathogenesis.Methods:96 BALB/C mice were divided into four groups.A is asthma model group,B is Budesonide group,C is Monlukast group,D is negative control group.All mice were actively sensitized by intraperitoneal injection of 0.5ml liquid(including 100ug ovalbumin and 400ug Al(OH)₃) on day 0 and day 7 except D group,From day 14 to day 21 the sensitized mice were challenged with an aerosol ovalbumin(1%w/v diluted in sterile physiological saline) daily for 30 minutes.The C group were given montelukast(6mg/kg) one hour after antigen challenge daily for 4 weeks,The B group were exposed to Budesonide suspl(including Budesonide 2ml and NS 3ml) for 30 minutes one hour after antigen challenge daily for 4 weeks,D group were conducted by i.p.injection of 0.5ml NS on day 0 and day 7,From day 14 to day 21 they were challenged with an aerosol NS daily for 30 minutes.Six mice were sacrificed in every group weekly for four weeks.The blood was obtained via excising eyeball as possible and put in the anticoagulation tube.the serum was achieved.The CC16 was measured via ELISA.Left lung were fixed with 10% phosphate-buffered neutral formalin and embedded in paraffin,sections were cut longitudinally,some of sections were stained with hematoxylin and eosin,and some of sections were performed immunohistochemical staining for CC16,the right lung was freezed and grinded,RT-PCR was performed for CC16mRNA. Results:1,CC16 concentration in peripheral blood between four groupsThe OD of CC16 in A,B,C groups was significantly decreased for 1,2and 3 wk as compared with this of control group(P＜0.05) and The OD of CC16 was not statistically significant between four groups for 4 wk(P＞0.05),The OD of CC16 in B,C groups was significantly decreased only for 1 wk as compared with those of A group(P＜0.05).The OD value of CC16 in A,B,C groups was significantly increased along with time from the 1^th week to the 4^th week(P＜0.05),but the OD value of CC16 in D group was not(P＞0.05).2,CC 16 coloring degree via immunohistochemical staining between four groupsThe coloring degree of CC 16 via immunohistochemical staining in A,B,C groups was significantly increased as compared with this of control group for 1,2,3 and 4 wk(P＜0.05); The coloring degree of CC16 via immunohistochemical staining in B group was significantly decreased as compared with this of A group(P＜0.05);there was statistically significant between A and C group only for 4 wk(P＜0.05);there was not statistically significant between C and B group only for 4 wk(P＞0.05);The coloring degree of CC16 via immunohistochemical staining in A,B,C groups was significantly decreased along with time from the 1^th week to the 4^th week(P＜0.05),but the coloring degree of CC16 in D group was not(P＞0.05).3,CC16mRNA expression between four groupsThe expressions of CC16 in A,B,C groups was significantly decreased for 1,2,3 and 4 wk as compared with those of control group(P＜0.05);The expressions of CC16 in B groups was significantly increased as compared with A group for every week(P＜0.05);The expressions of CC16 in C groups was significantly increased as compared with A group only for 1,4 week(P＜0.05) and was not statistically significant as compared with B group only for 1 week(P＞0.05).The mRNA expressions of CC16 in A,B,C groups was significantly increased along with time from the 1^th week to the 4^th week(P＜0.05),but the mRNA expressions of CC16 in D group was not(P＞0.05).4,Correlation between CC16 concentration in peripheral blood and CC16 mRNA expression in lungThe correlation between CC16 concentration in peripheral blood and CC16mRNA expression is totally positive correlation.(r=0.777,p＜0.0001).Conclusions:1,The expression of CC16 mRNA in lung tissue of asthmatic mice and CC16 concentration in peripheral blood are all decreased,and gradually reduced with the increase of the inflammatory cell in lung tissue.So CC16 concentration probably is an index of asthmatic airway inflammation.2,ICS(budesortide) can significantly increase the CC16 expression oft he asthmatic mice,and the effect that CC16 resist to the airway inflammation of asthma was significantly increased over time.3,Montelukast can significantly increase the CC 16 expression of the asthmatic mice after given 4 weeks,The effect of montelukast maybe same with that of budesonide after 4 weeks,although its effect is slower than that of budesonide in the first 3 weeks.4,Peripheral blood CC16 and CC16 mRNA expression in lung tissue were positively correlated,so it can not only reflect the lung tissue expression of CC16 mRNA in lung tissue,but also indirectly reflect the severity of asthma inflammation.