| Objective:This experiment is on the basis of the optimal proportion to treat asthma that previous animal experiments have confirmed the viewpoints that day moxibustion medicine white mustard powder acupoint sticking therapy,determine that the application of ancient prescription of day moxibustion medicine white mustard powder acupoint sticking therapy for asthma rats could significantly inhibition of airway inflammation and immune regulation,prompting immune function returned to normal,ancient prescription of day moxibustion medicine white mustard powder play an important role in the immune state reconstruction.Proposed from the perspective of gene white mustard powder in the pathogenesis of infantile asthma,the steady-state reconstruction immune mechanism,to study the therapeutic mechanism and so on,to improve the diagnosis accuracy and treatment of asthma targeting,and provide experimental basis for clinical application and theoretical support.Methods:90 clean male SD rats,4 ~ 6 weeks,weight(150 ±20)g,according to the equilibrium method of random stratified(by weight)is divided into three groups,respectively to medicine group,model group,group A and group B(15),the normal group,each group 30 experimental animals.Day 1,give a medicine group,model group,respectively with fresh preparation of OVA,100 mg with 200 mg of aluminum hydroxide gel and saline 1 ml of suspension liquid in rats after two groin,foot plantar,a total of 4 do hypodermic,each point of 0.2 ml,intraperitoneal injection of 0.2 ml at the same time,the normal group do not do any processing.15 th day of the execution group A model group,normal group,5 rats that need to be detected and save parts of the specimen collection,in order to confirm the building after A successful animal gene expression in the body of the state,model B group retention as A treatment for comparison.15 to 24 days to begin day moxibustion medicine group were applied treatment,acupoints were applied with 8%sodium sulphide in acupuncture points before hair removal,affix pill in medical desensitization gel,stick in the rat lung yu(double)respectively,anti-asthma,CV 17,bl 43(double),1 times a day,each time attach 2 ~ 4 h,applied after treatment,the rats of experimental group sensitization in an airtight container with ultrasonic atomizer atomized inhalation of 2% OVA saline suspension,30 min every day,to stimulate out of 1,10 daysin a row.The model group B was given a blank patch,and the treatment method was given to the drug group.The normal group is free to eat regular feed and water.The rats were sacrificed on the 25 th day to collect lung tissue samples.For RNA extraction and identification,gene chip detection,bioinformatics analysis,using Western blot,Northern blot,fluorescence quantitative rt-pcr detection and gene knockout technology of gene chip detection results for validation.Results:(1)The model successfully concluded that,compared with the normal group,the rats in the model group had significant changes in behavior,pathology and genetics,and the model was successful.(2)Due to limited funds,the genes extracted from lung tissues of 9 rats were examined,the genetic results based on data such as the gene pool,according to the p < 0.05,for gene expression times value the absolute value of 2.0 or higher threshold to identify differentially expressed genes.The results were statistically significant,and these genes were directly or indirectly related to the development of asthma.Group A is normal group and model group,normal group by 17 genes,raised six genes,instead of change appears in group A model group,was statistically significant,asthma model success.Normal group and model group,group B to group comparison,differentially expressed genes,the same raised gene has four,cut gene has 2,cut multiple gene dosage group was obviously higher than that of model group,group B raised gene and cut to decrease.Model group of group A and group B,the dosage group,raised gene has two,has six cut gene,to cut A multiple gene medicine group was obviously higher than that of model group,group B model raised in group A set of genes in the group B in the model group,medication group cut,cut or low expression of gene is not expressed in model group B,the dosage group appeared two raised genes,it has anti-inflammatory effects.In the comparison between group B and drug administration,there was no down-regulated gene expression,and there were 8up-regulated genes.The results indicated that group B of the model group had their own immune recovery,and the treatment effect was better than that of the model group B,and the treatment effect was not restored to normal.(3)To the normal group,model group and drug group to compare the differentially expressed genes,through heat maps can be found that the gene expression of normal ratsby a rise in different genes and steady-state,cut or not to maintain the immune model group and drug group of genes,there is raised and lowered to deal with the occurrence of asthma,the difference was statistically significant,found 18 raised genes associated with asthma,cut,there are seven genes.The experimental results were consistent with some published reports and were consistent with the function of genes.Conclusion:1.The differentially expressed genes and asthma: the relationship between the pathogenesis related to EOS,participate in the airway inflammation,airway hyperresponsiveness,airway remodeling process of genes are: EOS chemotaxis cytokines CCR3,Ccl11,Ccl2,Ccl22;Genes with anti-inflammatory effects: ADM,Pla2g4 c,Fcgr3a,Nr1d1,Acox1;The genes involved in airway inflammation were: C3,SelP,Sele,Serpina3 n,and CX3CR1.The genes involved in airway inflammation and airway remodeling were mmp-9 and TLR1.The genes for airway inflammation and airway remodeling are: Timp1;It is closely related to asthma,but it is not clear that the genes involved in the mechanism are prim1,Serpinb10,FMO3,Scd1,per3,DBP,ANKRD1,CD177,DES.2.Compared with the group A of the normal group and the model group,the gene was down-regulated,and the three mechanisms involved in the formation of asthma were related.The model of asthma was established successfully,and the difference was statistically significant.Normal group,model group,A group and model group compared with group B,the dosage group,the differentially expressed genes,the same dosage group cut ratio is higher than the model group,group B gene on the comparison between the two way,differentially expressed genes were all raised,no cut gene,said moxibustion medicine tomorrow after white mustard powder to treat asthma rats,compared with asthma model group in group A and group B situation improved,compared with normal group asthma inflammation still exist,the result was statistically significant.3.Under normal circumstances,the immune steady maintaining the basic state of life,of rats with asthma rats immune the breakage of the steady state,the genes associated with asthma also produces change,in the heat map clear cut on gene changes,should have raised genes,into the cut,cut genes into raising originally,originally not express or lower expression genes begin to express,the normal group,model group and drug group of differentially expressed genes were compared,found 18 raised genes associated withasthma,seven cut gene,through the comparison analysis of various genes,The experimental results agree with published some related literature reports,also consistent with the function of genes,on genes,asthma rats genetically changed,some of these genes or gene polymorphism in clearly play an important role in asthma,to treat asthma immune reconstruction process provides a direction of the steady state. |
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