Jia Wei Xiao Chai Hu Tang Intervention In Asthmatic Rat Model Of Experimental Research

Purpose:In the establishment of rat model of bronchial asthma,based on Xiao Chai Hu Tang Jia Wei given intervention,through animal experiments,from pathology,immunohistochemistry and molecular biology,perspectives of Xiao Chai Hu Tang Jia Wei of Chinese medicine treatment of bronchial asthma rats model of the mechanism ,for the treatment of asthma medication and provide experimental evidence.method:PartⅠ: The Histopathological observation of Xiao Chai Hu Tang Jia Wei in the rats with asthmaAfter feeding in the adaptation, 60 rats were randomly divided into 6 groups,namely the normal group, model group,Xiao Chai Hu Tang Jia Wei high dose group (the high dose group),Xiao Chai Hu Tang Recipe for low-dose group (low dose group),western dexamethasone injection group,Tonga, Western medicine Modified Xiaochaihu dexamethasone injection group (in western medicine group),10 in each group.The rats were marked on the numbers,the average weight difference between groups control <20g, to intervene with medicine.Treatment groups were from the 1st of provocation of asthma (modeling the first 3 weeks) to put to death the day before oral administration for 4 weeks,normal group with normal saline in each group review the animal body weight every 7 days.According to the report,he rats were kept three days after adaptation, in addition to the normal group, in experiment 1 and day 8 days the rats by intraperitoneal injection of 10% egg protein,the mixture of aluminum hydroxide 1ml, the waist on both sides depicting 1 point for every point injected 0.5 ml,a total of 2 ml, so that rats sensitized.The first 15 days only use 40 ml of 3% egg protein stimulate models of asthma spray, aerosol flow control in 5ml / min,each 15 minutes,once every other day,a total excitation for 4 weeks. In rats breathing speed up,cyanotic lips, nodded breathing and unstable performance of the excited standing success.Normal control group received normal saline intraperitoneal ovalbumin injection and aerosol inhalation.After the animals were anesthetized completely,the rapid ligation of right main bronchus chest,take right middle lobe of 4% paraformaldehyde, dehydrated,embedded,sliced,hematoxylin - EOSin HE staining,pathological changes were observed.Before modeling,measurement of body weight once;modeling every 7 days after the beginning of the first body weight measurement,drawn up before the last measurement.Before modeling, measurement of body weight once; modeling every 7 days after the beginning of the first body weight measurement, drawn up before the last measurement; rats stimulated within 1 minute after a four-count breathing rate and the number of sneezing.All data were analyzed with SPSS10.0 software processing.Experimental data with mean±standard deviation indicated that groups using single factor analysis of variance (Bonferroni) method for statistical analysis.The level of significant difference between the 0.05 and 0.01 as the standard.Part II: The influence of Xiao Chai Hu Tang Jia Wei on NO and ET-1 in asthmatic ratsAnimal groups, modeling,drug use and the statistical processing with the former.In the experiment 42 days after the last weighing excited asthma and blood of rats in the abdominal aorta for the determination of nitric oxide,endothelin -1.Determination of nitrate reductase by NO levels in lung tissue, NO chemical properties as active,in vivo metabolic soon to NO2-and NO3-,which is further converted to NO2-NO3- , nitrate reductase specific law used to NO3-reduction to NO2- , determined by color depth The level of concentration . Radioimmunoassay using ET-1 content in lung tissue.Radioimmunoassay by the automaticγ-counter pre-programmed to directly give the standard curve and sample concentrations.Part III: The influence of Xiao Chai Hu Tang Jia Wei on IL -4 and IFN-γof rats with bronchial asthma. Animal groups,modeling,drug use and the statistical processing with the former.Animal anesthesia completely after ligation of right main bronchus chest quickly,take right upper lobe, weighed and homogenized moved into the glass tube,add 2 times the weight of the lung tissue of saline, full ground up,so that tissue homogenization. Preparation of a good homogenate 3500r/min, centrifuge 10 min,the supernatant extracted separate packaging, low temperature refrigerator tested.Interleukin (IL) -4 andγ-interferon (IFN-γ) were measured by enzyme-linked immunosorbent assay (ELISA).Part IV: The influence of Xiao Chai Hu Tang Jia Wei on EOS of lung tissue and blood in the rats with asthmaAnimal groups,modeling, drug use and the statistical processing with the former. Animal anesthesia completely after ligation of right main bronchus rapid thoracotomy, take right middle lobe of 4% paraformaldehyde, dehydrated,embedded,sliced,hematoxylin - EOSin HE staining was observed in the EOS infiltration in lung tissue changes.Part V: The expression of Xiao Chai Hu Tang Jia Wei on IL-5 mRNA of lung tissue in the rats with asthmaAnimal groups,modeling,drug use and the statistical processing with the former. Animal anesthesia completely after ligation of right main bronchus chest quickly,take the right lung near the hilum Department about 100mg,into Eppendorf tubes,adding 1ml Trizol fully homogenized, put into liquid nitrogen tank,tested RT-PCR. Using RT-PCR method to detect in the lung tissue IL-5 mRNA expression.Results:1.The success of rat model of bronchial asthma.2.Bronchial asthma and lung tissue adjacent to a large number of EOS vascular infiltration,consistent with asthma,chronic inflammation and pathological changes.Xiao Chai Hu Tang Jia Weican reduce the high and low dose EOS infiltration in lung tissue, in particular, Xiao Chai Hu Tang Jia Wei EOS high-dose group decreased more significantly, better than the model group Xiao Chai Hu Tang Jia Wei low dose group.3.The changes in body weight of experimental rats:before sensitization, before stimulation,stimulate two weeks,four weeks each time point excitation, the excitation before and after the normal group and Xiao Chai Hu Tang Jia Wei low dose group,Xiao Chai Hu Tang Jia Wei high dose group, traditional Chinese medicine Modified Xiaochaihu Tonga Western dexamethasone injection group,dexamethasone injection group no difference in body weight (P> 0.05); stimulate body weight after two weeks was significantly lower than model group rats, high doses of Xiao Chai Hu Tang Jia Wei group, Tonga, Western medicine Modified Xiaochaihu dexamethasone injection group, the difference was significant (P <0.01);excitation model rats after 4 weeks body weight was significantly lower, Modified high-dose group Xiao Chai Hu Tang,Xiao Chai Hu Recipe for Tonga Western dexamethasone injection group (P <0.01),lower than the dexamethasone injection group (P <0.05).Rats in each experimental group the number of respiratory rate and changes in sneezing: excited after four weeks, compared with model group, model group, the number of respiratory rate and sneezing was significantly higher than other groups.4.Compared with the control group, model group,NO increased significantly (P <0.05),the other groups were not significantly different (P> 0.05). Compared with model group,Western group, high dose of Chinese medicine,Chinese and Western medicine group NO level was significantly lower (P <0.05). Western Group,Western Canada Chinese medicine group, low dose of Chinese medicine and Chinese medicine high dose group between the two groups were not significantly different (P> 0.05).Compared with the control group,model group,Western group,high dose of Chinese medicine,Chinese medicine group and the Western Canada Chinese ET1 low-dose group were significantly increased (P <0.01);with model group, Western group,the high dose group of Chinese medicine,Chinese and Western medicine Chinese medicine group and the low dose group had significantly lower levels of ET1 (P <0.01). Western medicine group,high dose group of Chinese medicine,Chinese medicine group and the Western Canada Chinese low-dose group ET1 levels between the two groups were not significantly different (P> 0.05).5.Compared with model group,modified Xiao Chai Hu Tang can significantly lower high-dose IL-4 levels, with significant difference (P <0.01); while the normal group, and flavored low-dose group Xiao Chai Hu Tang,Xiao Chai Hu Tang Jia Wei high dose group,Tonga,Western medicine Modified Xiaochaihu dexamethasone injection group,dexamethasone injection group rat lung tissue between the IL-4 levels not different (P> 0.05), Compared with model group, Xiao Chai Hu Tang Jia Wei high dose, low dose of Modified Xiao Chai Hu Tang Xiao Chai Hu Jia Wei Tonga group and western medicine can dexamethasone injection group was significantly higher IFN-γlevels, with significant difference (P <0.01 ),western group can also be elevated IFN-γlevels, but no significant difference (P> 0.05);with the normal group, the western group and model group,IFN-γlevels,which have a very significant difference (P <0.01 );compared with western medicine,Chinese medicine Modified Xiaochaihu Tonga dexamethasone injection group can be significantly increased IFN-γlevels, with significant difference (P <0.01), high doses of Modified Xiao Chai Hu Tang Xiao Chai Hu Tang Group and the Modified low-dose group also significantly increased IFN-γlevels, with significant differences.6.Compared with model group,Western group,high dose of Chinese medicine,Chinese and Western medicine EOS, lung bronchial tissue was significantly lower (P <0.01), low dose of Chinese medicine EOS,bronchial tissue was significantly lower (P <0.05).Western Group,Western Canada Chinese medicine group,low dose of Chinese medicine and Chinese medicine high dose group between the two groups no significant difference was not significant (P> 0.05). Compared with model group,Western group,high dose of Chinese medicine,Chinese and Western medicine group peripheral blood EOS counts were significantly lower (P <0.01), traditional Chinese medicine low dose group was significantly lower in peripheral blood EOS counts (P <0.05). Western Group,Western Canada Chinese medicine group,low dose of Chinese medicine and Chinese medicine high dose group between the two groups no significant difference was not significant (P> 0.05).7.Asthmatic lung tissue IL-5 mRNA expression was significantly higher than the normal group (P <0.01),compared with model group, high dose of Chinese medicine,Chinese and Western medicine lung tissue IL-5 mRNA expression was significantly lower (P <0.01),compared with western medicine, traditional Chinese medicine in high dose group,Western Canada Chinese medicine group,lung IL-5 mRNA expression was significantly lower (P <0.01),compared with the low dose of Chinese medicine,Chinese medicine high dose group,herbal medicine lung tissue IL-5 mRNA expression was significantly lower (P <0.01),high-dose group and Chinese medicine Chinese and Western medicine between groups in lung tissue expression of IL-5 mRNA was no significant difference (P> 0.05).Conclusion:1. Multi-point subcutaneous injection using ovalbumin (OVA) approach, the success of induced rat model of bronchial asthma.2.Rats in all treatment groups significantly improved the general state of bronchial asthma and lung tissue adjacent to a large number of EOS vascular infiltration,consistent with asthma,chronic inflammation and pathological changes.Xiao Chai Hu Tang Jia Weican reduce the high and low dose EOS infiltration in lung tissue, in particular, Xiao Chai Hu Tang Jia Wei EOS high-dose group decreased more significantly, better than the model group Xiao Chai Hu Tang Jia Wei low dose group,can be seen Xiao Chai Hu Tang Jia Wei inhibit EOSinophil infiltration, reduce airway inflammation,reduce airway hyperresponsiveness,as Xiao Chai Hu Tang Jia Wei effective to provide a favorable basis for the treatment of asthma.3.Xiao Chai Hu Tang Jia Wei significantly reduced the expression of NO in blood.High and low dose group of Modified Xiao Chai Hu Tang can lower blood NO levels,but no significant difference between the two. Xiao Chai Hu Tang Jia Weican be well in lowering blood levels of ET-1,thus inhibiting the development of asthma inflammation,prevent or inhibit airway remodeling.4.Asthmatic lung tissue IL-4 levels were significantly higher than the normal group. And the model group,the treatment group could reduce the content of lung tissue IL-4,IFN-γlevels increased, indicating Modified Xiao Chai Hu Tang can lower IL-4 levels in lung tissue, increased IFN-γlevels,which regulate immune function reduce the infiltration of airway EOS to control airway inflammation.5.Xiao Chai Hu Tang Jia Weican significantly reduce the external peripheral blood of asthmatic rats, the number of bronchial in EOS, reduce asthma animal model of airway inflammatory cell infiltration,and reduce asthma animal model of accumulation of inflammatory cells in peripheral blood .6.IL-5 involved in the pathogenesis of bronchial asthma and airway inflammation, Xiao Chai Hu Tang Jia Wei high dose group,Chinese and Western medicine group,lung IL-5 mRNA expression was significantly reduced,modified Xiao Chai Hu Tang can reduce the level of transcription IL-lung 5mRNA expression,to promote EOS apoptosis in the lungs and reduce the infiltration of inflammatory cells,thereby reducing airway inflammation, relieve airway spasm.