Study On The Effect And Mechanism Of Acupunture To Relax Asthma Airway Smooth Muscle

Objective:By observing the latency period of inducing asthma,pulmonary function,the changes of airway morphology and tissue ultrastructure,protein expression and gene level of ET-1 andβ2-AR,and the levels of ET-1,TNF-a,cAMP and cGMP in serum and alveolar lavage fluid（BALF）to explore the effect and mechanism of acupuncture relaxation asthma airway smooth muscle on asthmatic rats so that could provide a theoretical basis for the clinical application of acupuncture in the treatment of asthma.Methods:48 SPF-rated male rats were randomly divided into normal group（NC）,model group（AS）,acupunture the Feishu matched Dingchuan group（AAF）,acupunture the Kongzui matched Yuji group（AAK）,acupunture the Feishu matched Dingchuan and Kongzui matched Yuji group（AAFK）,and false acupunture group（AAJ）,8 rats in each group.The asthma models rat were sensitized by OVA,except for the NC group,which was replaced by physiological saline.Recording asthmatic time of rats,detecting pulmonary function,observing changes in the morphology and ultrastructure of smooth muscle of the airways of rats,detecting the expression of ET-1,β2-AR mRNA and protein in lung tissues and the levels of ET-1,TNF-a,cAMP and cGMP in serum and BALF.Results:（1）Comparison of incubation period of inducing asthma:compared with the NC group,the incubation periods of inducing asthma in each group was shortened（P<0.05）;compared with the AS and AAJ group,the incubation periods of inducing asthma in the AAFK group,AAF group and AAK group were prolonged（P<0.05）.And compared with the AAFK group,AAF group was shorten（P<0.05）,and there was no significant difference between the AS group and AAJ group,AAF group and the AAK group.（2）Changes in pulmonary function:compared with the NC group,RL was significantly increased in AS group and AAJ group,and Cdyn was significantly reduced（P<0.05）,there was no difference in the first RL between AAF group and AAK group,and the last three RL were increased（P<0.05）,and there was no significant difference in the first two groups of Cdyn,and the last two were reduced（P<0.05）;compared with the AS group,RL was decreased in AAFK group,AAF group and AAK group,Cdyn was elevated（P<0.05）;compared with the AAJ group,RL was decreased（P<0.05）in AAFK,AAF and AAK groups,and Cdyn was significantly increased（P<0.05）in AAFK,while both Cdyn was elevated（P<0.05）in the first three times and had no significant difference（P>0.05）in the AAFK group.Cdyn decreased（P<0.05）;there were no significant differences between NC and AAFK,AS and AAJ,AAF and AAK.（3）ET-1,TNF-a,cAMP and cGMP levels in serum and BALF:Compared with the NC group,the contents of TNF-a,ET-1 and cGMP in serum and BAALF in AS group and AAJ group increased significantly,cAMP decreased significantly（P<0.05）,ET-1 and cGMP levels in BAALF in AAFK group increased（P<0.05）,AAF group,The content of ET-1 in serum（P<0.05）and ET-1 and cGMP in BAALF increased（P<0.05）in AAK group,and compared with AS group and AAJ group,the content of TNF-a,ET-1 and cGMP in serum and BALF in AAFK,AAF group and AAK group decreased and cAMP content increased（P<0.05）,and there was no significant significance between AS group and AJ group,AAFK group,AAF group and AAK group（P>0.05）.（4）Comparison of gene expressions:Compared with the NC group,ET-1mRNA expression andβ2-ARmRNA expression decreased（P<0.05）in the AS group,AAJ group,AAF group and AAK group;compared with the AS group and AAJ group,ET-1mRNA expression decreased andβ2-ARmRNA expression increased（P<0.05）in the AAFK group,AAF group and AAK group;compared with the AAFK group,the AAF group,β2-ARmRNA expression decreased in AAK group（P<0.05）,and there were no significant differences between NC group and AAFK group,AS group and AAJ group,AAF group and AAK group.（5）Comparison of protein level:Compared with the NC group,ET-1 protein expression was increased andβ2-AR protein expression decreased in the AS group,AAJ group,AAF group and AAK group（P<0.05）;compared with the AS group and AAJ group,the expression of ET-1 protein in AAFK group,AAF group and AAK group was significantly reduced.β2-AR protein expression was significantly increased（P<0.05）,compared with AAFK group,ET-1 protein expression was elevated（P<0.05）andβ2-AR protein expression was not significantly different,and there was no significant difference between NC group and AAFK group,AS group and AAJ group,AAF group and AAFK group,AAF group and AAK group（P>0.05）.（6）the change of airway smooth muscle morphology and ultrastructural:compared with the NC group,the AS group and AAJ group had severe abnormalities in lung tissue,collagen fiber hyperplasia,smooth muscle significantly thickened,mitochondria significantly swollen,a large number of crest fractures and decreases,dense spots and dense bodies increased significantly,and the distribution was uneven;The lung tissue of AAK group was severely abnormal,the smooth muscle was significantly thickened compared with the NC group,slightly reduced compared with the AS group and AAJ group,the mitochondria were slightly swollen,the crest part was broken,the dense spots and dense bodies were reduced,the distribution was uniform,the rough endoplasmic reticulum was slightly dilated,and the surface ribosomes were locally degranulated.The median lung tissue of AAF group and AAFK group was abnormal,smooth muscle was slightly thicker than that of NC group,significantly reduced compared with AS group and AAJ group,mitochondria were slightly swollen,a small number of crest fractures,the number of dense spots and dense bodies was acceptable,the distribution was uniform,and the ribosomal on the surface of the rough endoplasmic reticulum was abundant,and no obvious degranulation was seen.Conclusion:Acupuncturing the Feishu matched with Dingchuan and Kongzui macthed with Yuji acupoints can prolong the incubation period of inducing asthma,reduce inflammatory cell infiltration,and reduce airway inflammation which the mechanism may be that the two groups of acupuncture points simultaneously regulate the two targets of ET-1 andβ2-AR,downregulate the expression of ET-1mRNA and protein,and upregulate the level ofβ₂-ARmRNA and protein to reduce the content of ET-1,increase the content of cAMP,reduce the level of cGMP to maintain the dynamic balance of the two,and protect the stability of the smooth muscle morphology and ultrastructure of the airway.The purpose of inhibiting smooth muscle hyperplasia and relaxing the spasm of smooth muscle of the airways to reduce lung resistance and control asthma.