| Objective:To observe the expressions of Thymic Stromal Lymphopoietin(TSLP) and Thymic Stromal Lymphopoietin receptor(TSLPR),and the effect of Budesonide(BUD),bacillus calmette-guerin(BCG) intervention in asthmatic mice.To analyze the relationship of TSLP/TSLPR and asthma to provide new ideas of prevention and treatment for asthma.Methods:1.Forty Kunming mice were randomly divided into four groups(10 mice each group)on average,including asthmatic group,BUD intervention group,BCG intervention group and control group.2.In asthmatic group,mice were sensitized on days 1,7 and 14 with injection intrapertoneally 0.01%OVA and 10%aluminum hydroxide 0.2ml,and challenged by inhaling aerosol 2%OVA 5ml once a day for 30 minutes from 21 days to 27 days.3.In BUD intervention group,mice were inhaled BUD normal saline solution 5ml(containing BUD 1 mg) one hour before challenged.4.In BCG intervention group,mice were treated with injection intracutaneously BCG 0.025mg on 5d,3d,1d before sensitization.5.In control group,mice were sensitized and challenged with normal saline instead of OVA.6.Bronchoalveolar Lavage Fluid(BALF),peripheral blood,lung tissues and spleens were collected from mice 24 hours after the last challenge.Sections of lung tissue were performed and stained with HE.The expressions of TSLP,TSLPR in lung tissues were evaluated by immunohistochemistry.The levels of IL-4 in BALF,TSLP in peripheral blood,TSLP and TSLPR in lung tissues and spleens were measured by Enzyme-linked Immunosorbent Assay(ELISA).The expressions of TSLP mRNA and TSLPR mRNA in lung tissues were detected by real-time polymerase chain reaction.Results:1.The mean optical density(OD) of TSLP positive cells of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were (0.277±0.009),(0.064±0.015),(0.160±0.009),(0.004±0.001) respectively.There were obvious difference among the four groups(P＜0.01).The mean OD of positive cells increased noticeably in asthmatic group compared with the control group(P＜0.01),but that in BUD intervention group and BCG intervention group was lower than in asthmatic group(P＜0.01).The mean OD of positive cells increased noticeably in BUD intervention group and BCG intervention group compared with the control group (P＜0.01),but that in BUD intervention group was lower than in BCG intervention group(P＜0.01).2.The mean OD of TSLPR positive cells of asthmatic group,BUD intervention group, BCG intervention group and the control group mice were(0.413±0.030), (0.040±0.004),(0.168±0.008),(0.006±0.001) respectively.The comparative results were similar to those of TSLP positive cells.3.The levels of IL-4 in BALF of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(244.200±4.601) pg/ml,(220.600±3.034)pg/ml,(224.900±2.930)pg/ml,(219.200±2.480)pg/ml respectively.There were obvious difference among the four groups(P＜0.01).The levels of IL-4 increased noticeably in asthmatic group compared with the control group(P＜0.01),but those in BUD intervention group and BCG intervention group were lower than in asthmatic group(P＜0.05).Compared with control group,the levels of IL-4 in BUD intervention group and BCG intervention group were higher,but there were no significant statistical significance(P＞0.05).The comparative results between the BUD intervention group and BCG intervention group there were no significant statistical significance(P＞0.05).4.The levels of TSLP in lung tissues of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(4.221±0.115)ng/ml,(3.899± 0.071)ng/ml,(3.8980±0.046)ng/ml,(3.621±0.129)ng/ml respectively.The comparative results were similar to those of IL-4 in BALF.5.The levels of TSLPR in lung tissues of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(17.160±0.157) ng/ml,(16.460±0.209)ng/ml,(15.740±0.273)ng/ml,(15.860±0.285) ng/ml respectively.The comparative results were similar to those of IL-4 in BALF.6.The levels of TSLP in spleens of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(5.467±0.074)ng/ml,(5.096±0.077)ng/ml,(5.101±0.161)ng/ml,(4.975±0.073)ng/ml respectively.The comparative results were similar to those of IL-4 in BALF.7.The levels of TSLPR in spleens of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(20.901±0.421) ng/ml,(18.280±0.159)ng/ml,(18.840±0.273)ng/ml,(17.930±0.156)ng/ml respectively.There were obvious difference among the four groups(P＜0.01).Compared with control group,the levels of TSLPR in BCG intervention group were higher(P＜0.05).The other comparative results were similar to those of IL-4 in BALF.8.The levels of TSLP in peripheral blood of asthmatic group,BUD intervention group, BCG intervention group and the control group mice were(1.611±0.204)ng/ml, (0.484±0.061)ng/ml,(0.166±0.045)ng/ml,(0.336±0.076)ng/ml respectively.There were obvious difference among the four groups(P＜0.01).The levels of TSLP increased noticeably in asthmatic group compared with the control group(P＜0.01).But those in BUD intervention group and BCG intervention group were lower than in asthmatic group(P＜0.01).BUD intervention group and BCG intervention group compared with control group,the difference was no significant statistical significance(P＞0.05).But the levels of TSLP in BUD intervention group were higher than in BCG intervention group(P＜0.01).9.The levels of TSLP mRNA in lung tissues of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(8.510±1.657)%, (1.756±0.518)%,(3.950±1.031)%,(0.999±0.204)%respectively.The comparative results were similar to those of IL-4 in BALF.10.The levels of TSLPR mRNA in lung tissues of asthmatic group,BUD intervention group,BCG intervention group and the control group mice were(8.601±1.847)%, (0.972±0.295)%,(3.236±0.538)%,(1.848±0.394)%respectively.There were obvious difference among the four groups(P＜0.01).The levels of TSLPR mRNA in BUD intervention group were lower than in BCG intervention group(P＜0.01).The other comparative results were similar to those of IL-4 in BALF.Conclusions:1.The levels of TSLP in peripheral blood,TSLP,TSLPR in lung tissues and spleens,TSLP mRNA,TSLPR,mRNA in lung tissues were all increased in asthmatic mice.TSLP and TSLPR play an important role in pathogenetic mechanism of asthma.2.BUD and BCG can markedly reduce the expressions of TSLP and TSLPR of asthmatic mice.we provided the theoretical evidence of BUD and BCG for preventing and treating asthma.3.TSLP and TSLPR play an important role in the development of asthma,and may be the potential therapeutic target for asthma. |
