Effect Of Acupoint TDD Application On Inter Leukin-4,Recombinant Interferon-γ And Airway Remodeling Of Guinea Pig With Bronchial Asthma

[Background]:Asthma,also called bronchial asthma,is a chronic inflammatory disorder of the airway involving various inflammatory cells,such as mastocyte, eosinophilic granulocyte and T-lymphocyte,which result in the hyperresponsiveness, spasm and narrowing of the airway.It is manifests as paroxysmal wheeze,chest distress and/or cough,which usually occurs at night and/or at early morning and may remit spontaneously or by treatment.Asthma has a high incidence,which is considered to be the world's most common chronic inflammatory airway disease,and is a great threat to public health according to the incomplete statistics.There are about 100-200 million patients of asthma in the world,and in China it affects 1-2%of the population.Data of the recent years show that the incidence of asthma is rising;hence asthma is becoming the key disease to be studied.According to the discovery,asthma manifests not only the inflammation,but also the remodeling of the airway,which is referred to the extensive changes of airway structure,involving the thickening of the airway wall,metaplasia of squamous cell in airway epithelial layer,proliferation of airway goblet cells and smooth muscle,fibrosis of airway epithelial layer and differentiation of mesenchymal cells,particularly myofibroblasts.A general consensus is that asthma is a type of allergy disease,especially related to IgE-dependent.The imbalance of Th1/Th2 leads to the morbidity of asthma.INF-γis a characteristic Th1 cytokine,while IL-4 is a characteristic Th2 cytokine.In recent years,to adjust the imbalance of Th1/Th2 in the treatment of asthma has become popular,and the adjustion of the level of IL-4 is the key link.Glucocorticoids is the first line drug of the treatment of asthma which can anti-inflammatory,anti-allergy, regulate the role of immune function.But its side effects are very obvious.This disease belongs to the category of "Xiao Zheng"(Wheezing Syndrome) in TCM.Its pathogenic factors include endopathic and exopathic aspects.The former refers to congenital defect,kidney Qi deficiency and weak constitution,the latter is related to invasion lung by exogenous pathogenic factors.Point Application therapy in the treatment of asthma has a unique effect and is being widely used in clinical practice now.But the dosage is still an old form,an immediate hand-made cream or ointment,Hence,the research developed a new dosage of point application, Transdermal Drug Delivery paste(TDD),which is based on the cold-roaring prescription of "Zhang Shi Yi Tong".It is a new form of natural moxibustion with high bioavailability and fewer side effects.Affixed TDD to Dazhui(DU 14),bilateral Feishu(BL 13) and Shenshu(BL 23) could warm the lung,dispel cold,strengthen the spleen and kidney,resolve phlegm and relieve asthma.[Objective]:To observe the effect of acupoint TDD application on the InterLeukin-4(IL-4),Recombinant interferon-γ,(IFN-γ) in serum and bronchial Pathomorphologic changes of experimental bronchial asthma guinea pig in order to supply evidence for the therapeutic mechanism study and expand application of the Acupoint TDD Application.[Method]:The guinea pigs were randomly divided into 4 groups:Negative group,Model group,Dexamethasone(DEX)injection group and TDD group.Animals were made of asthma model by intraperitoneal injection of 10%egg albumin and 1% egg albumin aerosol inhaling method except that of the Negative group.The steps were as follows:①Allergens:On the 4th day,the models were established by sensitization with interaperitioneal injection of 10%ovalbumin(OVA)1mt;②Induced asthma:On the 18th day,the models were challenged with nebulized inhalation of 1%OVA,30s per time and one time per day for 14 days.③Animals in the negative group received the sensitization and asthma induced are same to that of the other groups,but with saline instead of OVA.④Treatment:The animals of TDD Group received the treatment with the Acupoint TDD Application on the 18th day,1 time every other day.The Points included Dazhui(DU 14),bilateral Feishu(BL 13),Pishu(BL 20) and Shenshu(BL 23).The animals of DEX group were injected the dexamethasone 0.5mg/kg in abdominal cavity,the time and times of the treatment were same to TDD group.The animals in the rest groups were not received any interventions after the allergens.⑤On the 31st day,after the treatment,all the animals were anesthesia by 3%pentobarbital sodium and rapidly decapitated.Took 5ml blood from each animal and kept them under the room temperature for 2h,then centrifuged with 3000 turn / min for 15min and took the supernatant to test the level of IL-4 and INF-γ.The test was strictly performed according to the immunohistochemical method(ELISA).Took each 4 pieces bronchial organizations from every animal and observed the pathological changes under light microscope.⑥All experimental data were set up on SPSS 13.0 database and statistical analyzed with P＜0.01 and P＜0.05 as the standard statistical significance.[Result]: 1 The serum level of IL-4:Compared with that of the Negative group,the serum level of IL-4 of the Model group was obviously increased(P＜0.01);after treated by TDD or dexamethasone,they decreased obviously(P＜0.05),however,there was not any difference between the DEX and TDD groups in the decreasing of IL-4(P＞0.05).2 The serum level of IFN-γ:Compared with that of the Negative group,the serum levels of IFN-γof the other 3 groups were significantly lower(P＜0.01);The serum level of IFN-γof DEX group was significantly higher than that of TDD group and model group(P＜0.01);The serum level of IFN-γof TDD group was significantly higher than that of model group(P＜0.01).3 Pathomorphologic changes:Negative group:Bronchial epithelial was integrity, and smooth muscle was not obvious thicken,alveolar wall was clear integrity,spacing patterns was normal and there was not any obvious inflammatory cell infiltration; Model group:Focal chronic infiltration of inflammatory cells in submucous connective tissue of main bronchus,vascular congestion,hyperplasia of glands, diffuse chronic infiltration of inflammatory cells in alveoli of lung,narrowed lumen and consolidation of alveoli.DEX group:The bronchial epithelium was nearly integrity,smooth muscle layer was mild thick,less interstitial inflammatory cells,the alveolar space was integrity and infiltrated a small amount of inflammatory cells; TDD group:Bronchial epithelial was almost integrity,the smooth muscle layer was slightly thick,alveolar integrity which mild interstitial hyperemia had some inflammatory cells infiltrated.[Conclusion]:Acupoint TDD application,similar to dexamethasone injection, can decrease the serum IL-4 level and increase the serum IFN-γ,level,reduce inflammatory cells count in bronchi and lung of asthmetic guinea pigs,relieve the inflammatory reaction,decrease the thickness of airway intima and thickness of fiber structure in main airway and the thickness of smooth muscle in small airway to relieve the damage of ultrastructure,which may relate to its mechanism in relieving asthma.