Effect Of Acupuncture On Th17/Treg Balance In Chronic Obstructive Pulmonary Diseases Model Rats

Objective To investigate the regulation of acupuncture on lung tissue Th17/Tregbalance in Chronic Obstructive Pulmonary Disease(COPD) rats induced by cigarettesmoke and lipopolysaccharide.Methods1.Thirty male SD rats were randomly divided into3groups: controlgroup，model group，and acupuncture group;2. The model group and the acupuncturegroup rats smoked in third days before and smoked fifteenth days, with6%chloralhydrate anesthesia, cervical incision, the trachea, Solution1ml syringe to LPS intracheal injection concentration of1mg/ml (lipopolysaccharide, LPS),200μ g/rats.Thesmoke box was placed in rats,with this card36cigarettes its smoked,for the first40minutes daily smoked, lasted8weeks, Then for each smoked for30minutes for4weeks.A totle of weeks of smoking weeks. The pathological change in lung tissue andpulmonary resistance and compliance were used to evaluate the COPDmodel;3.acupuncture method:firstly6%chloral hydrate anesthesia;secondlyacupuncture acupoint Feishu(BL13) and Zusanli(ST36);thirdly connect pointselectroacupuncture instrument, electroacupuncture parameters2mA,4/20Hz alternatingfrequence,every30minutes, every other day,a totle of14times of electroacupuncture.model group and control group was performed for6%chloral hydrate anesthesiaproperly. At the same time the model group and acupuncture group were all moderatesmoked;4.All rats were sacrificed after four weeks，ELISA method determinate thecontect of IL-17、IL-10、INF-γ、and TNF-α in plasma and bronchoalvedar lavagefluid,and calculate the totle number of white blood cells in BALF under lightmicroscope;5.Immunohistochemical method to observe the expression of IL-17R andIL-10R in lung tissue；6.Western blot method to observe the expression of IL-17R inlung tissue;6.Detection of IL-17and IL-10mRNA with Real-Time quantitativePCR.7.Analysis of relationship between IL-17、IL-10and lung function index. Results1. Compared with control group, the rats in the model group is significantly increased inthe resistance of pulmonary ventilation （P<0.001）,and dynamic lung compliance issignificantly decreased（P<0.001）.It shows that the model rats had strongly breathlimited. Compared with model group, the rats in the model group is significantlydecreased in the resistance of pulmonary ventilation（P<0.01）,and dynamic lungcompliance is obviously increased（P<0.05）， It shows that acupuncture can obviouslyimprove the pulmonary ventilation function.2. Normal lung tissue is normal; thickening visible lung interval of the rats in the modelgroup，inflammatory cell infiltration，small airway lumen rare secretion，alveolareduced， alveolar cavity expansion even fusion. Acupuncture group pulmonarythinning，alveolar area increased，alveolar interstitial inflammatory reduced，airwaysecretion decreased.3. ELISA results shows that，compared with control group, IL-17、TNF-α and INF-γ in rats of model group is obviously higher,compared with model group,acupuncture group was obviously lower（P<0.05）. Compared with control group, IL-10in rats of model group was obviously lower（P<0.05）, compared with model groupacupuncture group was obviously higher（P<0.05）.4. Immunohistochemistry shows that，IL-17R is mainly distributed in the lung andairway inflammatory cells. Optical density analysis shows that, compared with controlgroup,the IL-17R in model group is significantly higher（P<0.01）; compared withcontrol group, the acupuncture group is obviously lower（P<0.05）.5.Western blot and fluorescence quantitative PCR results shows，compared with controlgroup,IL-17R protein and mRNA expression of rats in the model group are significantlyhigher（P<0.05）. compared with control group, the acupuncture group are obviouslylower（P<0.05）. 6. IL-17is positively correlated with pulmonary resistance（r=0.609, P<0.01）；while isnegatively correlated with pulmonary dynamic compliance（r=-0.549，P<0.01）。7. IL-10is negatively correlated with pulmonary resistance（r=-0.532, P<0.01），while ispositively correlated with pulmonary dynamic compliance（r=0.512，P<0.05）.Conclusions Acupuncture may down regulate IL-17production, the formation of theup regulation of IL-10,which regulate the Thl7/Treg immune imbalance in rats with,inhibit inflammation, alleviate airway remodeling, and improve the Pulmonaryventilation function.