Based On The Regulation Of Transcription Factors And Theirgene Expression To Explore The Mechanism Of Vesiculation On Preventing And Controlling Asthma

ObjectiveThrough the establishment of bronchial asthma rat model, from the observation of rat asthma symptoms, airway tissue pathology and bronchial morphology, plasma and lung tissue homogenates of each animal of eosinophils (EOS) immunoglobulin E count and lung tissue homogenate (IgE), cytokine interleukin4(IL-4), and interferon gamma (IFN-γ) transcription factor T-bet in lung tissue, content of (T-box express in T cell) and GATA-3(GATA binding protein3) the expression of proteins, are observed from the quantity effect, time effect relationship, the objective evaluation of the efficacy of prevention and treatment of Asthma Day moxibustion, moxibustion for prevention and treatment of asthma.Methods70SPF class male SD rats were randomly divided into seven groups, were divided into normal group, placebo group, dexamethasone group, bigger long-range medicine group, bigger, cake short-range medicine group, long-range plaster group, short-range medicine group,10rats in each group. First days and seventh days of intraperitoneal injection of lml OVA/AL in the experiment (OH)3suspension [containing10mg OVA and200mg AL(OH)3] sensitization, fifteenth day rats were placed in the self-made atomization aspiration box, use10mg/L OVA saline solution inhalation excitation, continuous excitation two weeks in rats shortness of breath, wheezing and other symptoms. In addition to the normal group with physiological saline atomization inhalation, while other groups with ovalbumin (OVA) molding sensitization method, after the success of modeling, the rats the nape of the neck to hair, to day moxibustion on medicine group, the large group with lcm x lcm size plaster medicine cake, cookies, medicine group with the medicine cake0.5cm*0.5cm size plaster, specific acupoints refer to China Development, animal feeding at the temperature of36℃-37℃, similar V weather temperature and humidity75%indoor, the preparation of drugs by first days posted:Dingchuan, Feishu, Fenmen; second days posted:Feishu, Jueyinshu, Gaohuangshu; third days posted: Feishu, Pishu, Shenshu, fourth days:Feishu, Pishu, Shenshu, each5hours, one times a day, four times for a course of treatment, and then continue to conventional breeding, rest three day after one course of treatment to the above method, and so on, long course of treatment group were treated for three courses after the treatment, the same method short-range medicine application group one courses of treatment. The placebo group with the flour into a paste on the nape of the neck with rats, treatment and long term medicine group, dexamethasone group were given dexamethasone0.5mg/(kg·d) intraperitoneal injection, treatment with the long-range medicine group. All animal were at the same time, the treatment is finished, the last again after treatment with10mg/L OVA saline solution inhalation excitation was observed in all groups, animal airway symptoms, signs and record. Observation of symptoms after the rats were anesthetized, The rats were anesthetized, extraction of abdominal aortic blood for EP pipe. Remove the lung tissue, the left lung with pre cooling PBS cleaning and reserved, used for RNA extraction. Middle lobe of the right lung pathological sections for HE staining, optical microscope to find the bronchioles cross-sectional, complete image acquisition, analysis and calculation of standard pipe wall thickness, the standard smooth muscle thickness measurement. Weighing100mg right lung, lung tissue homogenate of the rest, is placed in the container, the ice bath tissue, cryopreservation standby. EOS blood analyzer, IgE, IL-4and IFN-Y levels by enzyme-linked immunosorbent assay (ELISA) detection, real-time fluorescence quantitative (RT-qPCR) assay, the expression level of T-bet mRNA transcription factor GATA-3mRNA.Results(1)The general situation in rats:The rats in the normal group expression of the spirit of lively, responsive, breathing, coat color is bright and clean, without any a shortness of breath, difficulty or wheezing. The rats in the model group shortly after inhalation of OVA appeared to be restless, scratching the face fur and other allergic symptoms, then rats fell down motionless, shortness of breath, abdominal breathing, cyanosis, some rats may hear and sonorous wheeze during expiratory phase, the nose outflow mucus. Repeatedly stimulated rat dull coat, depressed and unresponsive, dexamethasone group from the beginning that drinking water increased, rapid weight loss, black etc.(2) Moxibustion on asthma symptoms in rats:compared with the normal group, placebo group asthma incubation period was shortened, the number of fall rate, sneezing, itchy nose nasal scratching, nodding motion frequency increased significantly, the difference was statistically significant (P<0.05); Dexamethasone group, long distance medicine group compared with the placebo group, the asthma incubation was prolonged, the number of fall rate, sneezing, itchy nose nasal scratching, nodding motion number decreased significantly, the difference was statistically significant (P<0.05); And the short stick medicine group, long-range medicine group, dexamethasone group, asthma incubation period was prolonged, the number of fall rate sneezing, nasal itching, nasal scratching, nodding motion number decreased significantly, the difference was statistically significant (P<0.05); The same period, the incubation period of asthma medicine cake the size, the number of fall rate, sneezing, itchy nose nasal scratching number, point head movement of rats had no significant difference (P>0.05); Long distance medicine group and dexamethasone group, rat asthma incubation period, the number of fall rate, sneezing, itchy nose nasal scratching number, nodding movement has no significant difference (P>0.05).(3)Moxibustion effect on bronchial microscope pathology:The placebo group rat bronchial wall thickness and smooth muscle thickness was significantly higher than that in the normal group, the difference was statistically significant (P>0.05); Long distance medicine group, dexamethasone group rat bronchial wall thickness and smooth muscle thickness of the treatment group were significantly shorter on decreased, the difference was statistically significant (P>0.05); Long distance medicine group and dexamethasone group, rat bronchial wall thickness and smooth muscle thickness showed no significant difference (P>0.05); The same period, there is no significant difference of medicine cake size on rat bronchial smooth muscle thickness of the tube wall thickness change (P>0.05).(4) Effect of cold moxibustion on asthma rats EOS, LYM change in the placebo group:Rat peripheral blood and lung tissue EOS, LYM higher than that in normal group, the difference was significant (P<0.05); Compared with the long range medicine group, dexamethasone group and the placebo group, the rat peripheral blood and lung tissue EOS, LYM were decreased,, the difference was significant (P<0.05); Long distance medicine group and dexamethasone group in peripheral blood and lung bronchial tissue EOS, similar to LYM, no significant difference (P>0.05); Long distance medicine group, dexamethasone group and short medicine group, rat peripheral blood and bronchial tissue EOS, LYM decreased significantly (P<0.05); The same course, pie medicine group and little medicine group were not significantly different (P>0.05).(5)Effect of cold moxibustion on cytokines in lung homogenate of rats: compared with the normal group, the expression of IFN-γ placebo group decreased significantly, IL-4, IgE expression was significantly increased, IFN-γ/IL-4ratio was significantly decreased, the statistical significant difference (P<0.05); Compared with the placebo group, expression long distance medicine group and the dexamethasone group IFN-γ increased significantly, the expression of IL-4, IgE were significantly decreased, IFN-γ/IL-4ratio increased, the statistical significant difference (P<0.05);And the short stick group, the expression of long-range medicine group and the dexamethasone group IFN-γ increased significantly the expression of IL-4, IgE, decreased significantly, IFN-γ/IL-4ratio increased, the statistical significant difference (P<0.05); Long distance medicine group and dexamethasone group, no significant difference of IFN-in lung tissue homogenate gamma, IL-4, IgE change (P>0.05);No significant differences with the course change medicine cake the size of the IFN-of lung tissue homogenate in gamma, IL-4, IgE (P>0.05).(6)T-bet mRNA,GATA-3mRNA and the ratio of T-bet/GATA-3in the lung tissues of rats with placebo in the expression of T-bet mRNA:compared with normal group decreased significantly, while GATA-3mRNA expression increased, T-bet/GATA-3mRNA ratio decreased significantly, the differences were statistically significant (P<0.05); Long range and short range of paste paste in treatment group was significantly enhanced the expression of T-bet mRNA medicine group and the dexamethasone group, but the expression of GATA-3mRNA was decreased, T-bet/GATA-3mRNA ratio also increased significantly, the differences were statistically significant (P<0.05); Long distance medicine group and dexamethasone group, no significant difference was found in T-bet mRNA, GATA-3mRNA and T-bet/GATA-3ratio (P>0.05); There is no significant difference of the same course of medicine cake the size of the rat T-bet mRNA, GATA-3mRNA and T-bet/GATA-3ratio (P>0.05).Gonclusion(1)Moxibustion can relieve the inflammatory cell infiltration of airway, airway epithelial necrosis improved, can reduce the airway of asthma rat duct wall thickness, smooth muscle, can improve the airway inflammation and airway remodeling in asthmatic rats, and dexamethasone rather.(2)Moxibustion can reduce rat asthma symptoms and prolong the incubation period, decrease the rate of fall, sneezing, nasal itching nose, nodding movement. With the passage of days moxibustion, rats significantly reduce airway inflammation of asthma rat bronchial mucosa and submucosal apparent decline in the number of EOS and LYM bronchial lung tissue local infiltration of inflammatory cells, smooth muscle hypertrophy, edema, luminal stenosis, such as pathological changes significantly decrease, the tip day moxibustion therapy for asthma inflammation intervention is particularly important that day moxibustion has obvious timeliness, but with the size of the treatment medicine cake in this experiment for rat asthma symptoms and no significant pathological changes of the airway.(3)Moxibustion can significantly improve the low in asthmatic rats IFN-γ and T-bet mRNA expression level, reduce the amount of IgE, IL-4, GATA-3mRNA and EOS, so as to reduce the inflammatory cell infiltration in lung tissue of bronchus, and its mechanism may be related to induction of lung tissue T-bet mRNA expression was increased, inhibition of GATA-3mRNA expression, thereby regulating cytokine IgE, IL-4levels, reduce, increase IFN-γ, and ultimately reduce the number of peripheral blood EOS, thereby reducing the inflammatory cell infiltration in lung tissue of bronchus.