Study On Effects Of Moxa-Cone Moxibustion At Lung's Back-shu And Front-mu Points On Th17/Treg Balance In Asthmatic Mice And Its Mechanism

Objective: By observing the effects on the incubation period of inducing asthma,the histopathological changes of lung tissue,the expression of specific transcription factors PI3 K,Foxp3,ROR ? t m RNA in lung tissue,and the Th17/Treg percentage in spleen tissue of asthmatic mice after using moxa-cone moxibustion at lung's back-shu and front-mu points,discuss the mechanism of this therapy on treating asthma,and provide some experimental basis for preventing and controling asthma in clinical application.Methods:(1)Grouping: 48 male Balb/c mice were randomly devided into the normal group,model group,LY agent group,moxa-cone moxibustion group,each group of 12 mice.(2)Modeling: Mice in model group,LY agent group and moxibustion group were sensitized by intraperitoneal injection of ovalbumin(OVA)+aluminum hydroxide mixture,and stimulated by 1%OVA solution.The normal group was sensitized and stimulated by normal saline.(3)Intervention:Mice in LY agent group were injected the LY294002 into their caudal vein,and those in moxibustion group were taken moxa-cone moxibustion directly at their lung's back-shu and front-mu points.Mice in model group were fastened with the same way and same time as moxibustion group,while those in normal group had no any therapy.(4)Observation indexes: The incubation periods of inducing asthma were recorded,and the histopathological morphological changes of lung tissues were observed,and the percentages of Th17/Treg cells in spleen tissues were measured by flow cytometry,and the expressions of m RNA of PI3 K,ROR?t and Foxp3 in lung tissues were detected by real-time quantitative PCR.Results:(1)Comparison of the incubation periods of inducing asthma:Compared with the normal group,the incubation periods of inducing asthma in model group,LY agent group and moxibustion group were significantly shortened(P<0.01).Compared with the model group,the periods in LY agent group and moxibustion group were significantly prolonged(P<0.01).There was no significant difference between LY agent group and moxibustion group(P>0.05).(2)Comparison of histopathological morphological changes:The normal group had intact structure of lung tissue without damage,with no infiltration of inflammatory cell and no exudate in the bronchial cavity and alveolar cavity seen.In the model group,structure of the lung tissue was significantly damaged,and the cilia of bronchial mucosa cells was prostrate and lost,and even some cells were necrotic and shed.A large amount of exudate was observed in the bronchial lumen,and some lumen was narrowed and blocked.There were also hyperemia,exudation and even consolidation in the alveoli.Numerous inflammatory cells are seen around the bronchi and alveoli.LY agent group and moxibustion group were showed the similar manifestations,that lung inflammation was mild,and the structure of bronchi and alveoli was basically normal,as less exudation in the bronchial and alveolar cavity and less inflammatory cell infiltration were found.(3)Comparison of Th17/Treg percentages: Compared with the normal group,the percentage of Th17 cells in the spleen of the model group increased significantly(P<0.01),the percentage of Treg cells decreased(P<0.05)and the ratio of Th17/Treg increased(P<0.01).Compared with the model group,both LY agent group and moxibustion group showed the trends that the percentages of Th17 cells decreased(P<0.05,P<0.01),the percentages of Treg cells increased(P<0.01,P<0.05)and the proportion of Th17/Treg was down-regulated(P<0.01).There was no significant difference between LY agent group and moxibustion group(P>0.05).(4)Comparison of specific transcription factors: Compared with the normal group,the expressions of PI3 K and ROR ? t m RNA in the model group significantly increased(P<0.05,P<0.001),while the expression of Foxp3 m RNA significantly decreased(P<0.05).But compared with the model group,the expressions of PI3 K and ROR?t m RNA in LY and moxibustion groups significantly decreased(P<0.05,P<0.001),and the expression of Foxp3 m RNA significantly increased(P<0.01).There was no significant difference in those three indicators between moxibustion group and LY agent group(all P>0.05).Conclusions: Moxa-cone moxibustion at lung's back-shu and front-mu points can prolong the asthmatic mice's incubation period of inducing asthma and reduce the pulmonary inflammation.Its mechanism may be though regulating the PI3 K signaling pathway,affecting the expressions of specific upstream transcription factors ROR ? t and Foxp3,and intervening the secretion of Th17 and Treg cells,so as to correct the imbalance of Th17/Treg,improve the immune function of the body,inhibit the airway inflammation,and achieve the purpose of preventing and treating asthma at last.