The Establishment Of Northwest Dryness Syndrome Animal Model And The Intervention Trial By Empirical Formulas

Objective: to design the northwest dryness syndrome animal model,the macroscopic syndrome differentiation index and experience formulas disproof evaluation model,at the same time,on the model animals with partial index for testing and analysis of modern medicine,the northwest dryness syndrome and its correlation disease mechanism of modern medicine and drug prevention study deeply to the microscopic level.Methods: 1)The system summary since northwest dryness syndrome concept is put forward to carry out the research work,from the medical concept of overall,analysis of the multiple causes of northwest dryness syndrome,combing the complexity of northwest dryness syndrome syndrome structure,so as to reveal its pathogenesis essence,establish the basic treatment of northwest dryness syndrome and treatment,the formulas for the northwest dryness syndrome syndrome model study to lay a theoretical basis.2)On the basis of northwest dryness syndrome etiology,symptoms and the biological characteristics of common laboratory animals,choose suitable for the northwest dryness syndrome animal model research with strain.3)Draw lessons from the six evils of the past literature of animal building experience,based on the northwest dryness syndrome causes the results of the study,using its dry fire wind chill with actual load factor relationship between climate index of reference from the northwest dryness syndrome prone areas of hotan actual statistics by climate dry fire wind chill constituent ratio,and selected laboratory artificial climate index and classification,composition,quantitative and combined with time and space transform regulation of artificial climate index factors,establish experimental scheme.4)Analyze the four corresponding relationship of the clinical and animal representation of northwest dryness syndrome from the body(metrology)syndrome differentiation and syndrome differentiation to make the transition to animal syndrome differentiation,to establish the northwest dryness syndrome animal measurement dialectical method.5)Experimental animals were randomly divided into normal group,model group and treatment group the formulas for eight gasification cycle(a cycle is 6d)the causes of simulated building.6)With the northwest dryness syndrome animal differentiation of measurement scale as the main evaluation index model,choose water,diet and stool properties,weight combined with disproving evaluation of animal models,the formulas and establishing building gasification cycles required.7)Animal models with optical microscope,lung,submandibular gland tissues,lung,animal models with transmission electron microscope,submandibular gland tissue cell structure,evaluation of representative organs in building element under the action of damage degree.8)Using flow cytometry test peripheral blood T lymphocytes in the change in the process of building,suggests that during the period of building the dynamic change of T lymphocyte subsets.9)TUNEL method in detecting lung experimental animal representative organs and the status of the submandibular gland tissue cell apoptosis and evaluation mode that the damage degree of factors on the experimental animals.10)Immune histochemical method in detecting lung experimental animals and submandibular gland apoptosis related proteins the BCL-2,the expression of Bax,fas-l,preliminary explore the possible pathway of apoptosis.11)ELISA method to detect peripheral blood and promote experimental animals adrenal hormone(ACTH)and Cortisol(Cortisol),catecholamine(CA)content,evaluation of building factors of neuroendocrine immune network HPA axis.12)Associates method is used to determine serum malondialdehyde(MDA)concentration and xanthine oxidase method determining superoxide dismutase(SOD)activity,evaluation of building elements.13)With flow cytometry test peripheral blood T lymphocytes in the building process to experience the change of the treatment,the formulas that experience effect on T lymphocyte subgroup animal models with the formulas of dynamic change.14)With "dry,fire,wind,cold," gasification ark(i.e.,the northwest dryness syndrome main building factors)joint lavage thyroxine method to establish the northwest dryness syndrome animal model of heart and kidney Yin deficiency,flow cytometry test peripheral blood T lymphocyte by change,and the factorial design makes a research on the forming die factors influence of fT3,fT4.15)"dry,fire,wind,cold," ark of gasification combined egg albumen sensitization legal system for northwest dryness syndrome and bronchial asthma model,and ELISA method to detect serum IgE experimental animals,IL-4,IFN-gamma level,evaluate the role of two factors and experience formulas.Results: 1)team in the past study revealed a northwest dryness syndrome is the external cause to dry,fire,wind,cold,internal cause in dry sensitive physique condition,and appear inside and outside the hair dry permit and multifarious syndrome;According to the principle of dialectical treatment selected northwest dryness syndrome main northwest dryness syndrome and heart and kidney Yin deficiency syndrome and the northwest dryness syndrome experience formulas of bronchial asthma.2)The selected temperature,humidity,light,wind,dust and other five factors for building artificial climate indicators,and used a modified artificial climate box,adding dust experiment box,at the same time as the five artificial climate index classification quantification assignment to the combination and dry fire wind cold properties,and formulate gasification cycle state formula,artificial climate index factors in time and space transform regulation plan,select SD rats for northwest dryness syndrome main animal model building.3)compared with control group,model group rats after building factors exert namely appear the phenomenon of water quantity to increase,and continues until the end of treatment.In dealing with the three gasification cycle before eating less obvious,however,as the processing of applying,to 8d,model group diet is no statistical difference compared with normal control group.Compared with control group,model group rats weight at the beginning of the applying process of 4~6d weight slow growth,P=0.001(P<0.05)difference is statistically significant,continue to 10~12d;After 13 d and model group compared with the control group weight differences were not statistically significant.Compared with the control group,model group rats during building factors exert always have constipation.4)The pathological tissue section HE dyeing,24 d and lung and submandibular gland tissue of previous models did not see obvious pathological changes,48 group d pathological damage of individual animals began to appear more apparent.5)Peripheral blood cell count,according to the results of each experimental animal blood count within the normal range,compared with normal control group,24 d,elevated monocytes ratio(Mon %),percentage of eosinophils(Eos)% lower statistically significant.6)From the main syndrome animal model of peripheral blood T lymphocyte by the changes can be seen,when compared with normal control group: CD3+,CD4+,CD8+cells percentage change over time,but no statistically significant difference,the CD4+/CD8+ratio decline over time,6d differences statistically significant(P=0.019),shows that building factors can reduce the CD4+/CD8+ratio.7)Experimental animal lungs observed under transmission electron microscope and submandibular gland tissue structures in the cell,the model group,a significant phenomenon of apoptosis,cell membrane damage,cell swelling,alveolar visible between eosinophil infiltration.8)TUNEL experiment result shows model group of alveolar,around the bronchi and submandibular gland serous cell nuclei have obvious yellow to brown staining of apoptotic cells.9)Immune histochemical method detecting experimental animal lungs and submandibular gland apoptosis related proteins expression: model group the BCL-2 expression is not obvious,bax,fas-l have more expression,that building factors may result in promoting apoptosis protein expression level.10)ELISA to detect peripheral blood serum ACTH,Cortisol,and CA content in experimental animals,compared with normal control group,the model of rat serum adrenocorticotropic hormone(ACTH)content in indifference,and Cortisol(Cortisol)content decreased(P=0.020),catecholamine(CA)content is also reduced(P=0.019).Compared with model group,low,medium and high doses of the drug group can improve the serum catecholamine(CA)content(P=0.028,P=0.028,P=0.000).At the same time low,medium and high doses of the drug group can improve the level of Cortisol(Cortisol)levels back to normal,and the model group differences statistically significant(P=0.007,P=0.007,P=0.000).Medium and high doses of the drug can increase the content of ACTH,compared with the model group with statistical significance(P=0.034,P=0.034).11)Compared with normal control group,model group rats serum IgE levels,give a low dose and dose after the formulas in the lower-and there was no difference in the normal control group,high dose drug group rats serum IgE not seen is reduced,to higher levels compared with normal control group,with statistical significance.Compared with normal control group,high dose group can reduce the MDA content in serum.Compared with normal control group,low,middle dose group can enhance the vitality of SOD,no statistical difference and high dose group and normal group.Building factors can increase the expression of IgE,low,medium dose experience can reduce the expression,the formulas and high dose effect is not obvious.Experience in high doses can reduce MDA content,the formulas and low,medium dose can improve SOD vigor.12)From the main syndrome animal model for groups of peripheral blood T lymphocyte by the changes can be seen,when compared with 0d: the rat model with master card to medicine group peripheral blood CD3+cells percentage increases,statistically significant(P=0.002).CD4+cells percentage increases with time after the first drop,the difference of 6d statistical significance(P=0.000);Higher percentage of CD8+cells at 24 d,difference have statistical significance(P=0.014);CD4+/CD8+ratio change with time after the first drop rose,6d differences statistically significant(P=0.004),24 d statistically significant difference(P=0.013);The total number of lymphocytes no statistically significant difference in change over time.Compared to peripheral blood T lymphocyte by main syndrome animal model of change,the empirical formulas made mold adjustable factors of CD4+/CD8+ratio decreased.13)The northwest dryness syndrome of heart and kidney Yin deficiency of experimental animals peripheral blood T lymphocyte count: compared with normal control group,and the model group rats peripheral CD3+cells percentage to fall after rise over time,24 d difference have statistical significance(P=0.03);CD4+cells percentage increases with time after the first drop,no statistical significance difference with the normal control group,but 6d to 24 d difference statistical significance(P=0.042);Percentage of CD8+cells when the change is not statistically significant;CD4+/CD8+ratio drop change over time,6d differences statistically significant(P=0.004).Total number of lymphocytes fall after rise over time,including 0d and 6d differences statistically significant(P<0.05).14)The factorial design analysis of Yin deficiency made die factors with northwest dryness syndrome factors,with the change of FT3 and FT4,the results of variance analysis showed that the interaction of the two factors and two factors of the effect of free T3 content in the serum of rats with no statistical significance(F=0.752,P=0.752).And Yin deficiency building factors influence on free T4 content was statistically difference(F=5.592,P=5.592),the main card building factors influence on free T4 content no statistical differences(F=3.618,P=3.618),the interaction of the two factors on the impact of free T4 content in serum of rats was statistically significant(F=26.413,P=26.413).15)Compared with normal control group,the northwest dryness syndrome and bronchial asthma with pure bronchial asthma group make serum IgE levels,while no statistical differences compared to medicine group and normal group.Compared with normal control group,the northwest dryness syndrome and bronchial asthma with pure bronchial asthma group had elevated levels of serum IL-4,to medicine group and model no statistical difference compared with normal group.Serum levels of IFN-gamma no significant differences between groups.Conclusion: 1)Northwest dryness syndrome is square performance significantly,mainly dry evil cause,pathogenesis of involvement,how dirty mixed complex syndromes.2)The time and space in combination with artificial climate simulation experimental design can objectively reflect the northwest dryness syndrome scientifically exogenous dryness of the actual situation of fire wind cold causes made SD rat animal model conforms to the theory of traditional Chinese medicine,is feasible.3)The northwest dryness syndrome building factors exert the best time for four gasification cycle(d)24.Water quantity increased significantly during this period,the animal model,eating less,slow weight gain;Model animal lungs and submandibular gland has no obvious pathological changes.4)Further the microscopic index observation,northwest dryness syndrome main building factors can cause:(1)the classification of peripheral blood cell count is normal,the CD4+/CD8+ratio decline over time.(2)the cell membrane were observed under tem cell ultrastructure of visible damage,eosinophil infiltration and cell apoptosis.(3)animal models with TUNEL detect lung and submandibular gland apoptosis expression,and to promote apoptosis proteins bax immunohistochemical method to detect and high expression of fas-l.5)The northwest dryness syndrome model experience of the formulas was given from the experiment result shows:(1)the northwest dryness syndrome main building HPA axis is disorder,can cause animal model and the experience of high,medium and low dose the respectively the formulas have certain adjustment.(2)low,medium dose experience can reduce serum IgE level model animals,the formulas of high dose experience can reduce the MDA content in serum,the formulas at the same time,low dose can improve SOD vigor,showed that experience formulas have allergy,antioxidant,the ability of scavenging free radicals,and its role is associated with drug concentration.(3)the northwest dryness syndrome main building can cause animal model of CD4+/CD8+ratio decline over time,and after dosing(dose)can make the downward trend is obvious.6)Build the northwest dryness syndrome,heart and kidney Yin deficiency model preliminary study the main syndrome and building factors and interactions between building factors.7)Establishment of northwest dryness syndrome and bronchial asthma model,experience can reduce serum IgE animal models with the formulas,IL-4 levels,experience that the mechanism of treating bronchial asthma with the formulas,partly by reducing the synthesis of IgE,IL-4 and reach.