白芥子散穴位贴敷对哮喘大鼠气道炎症的影响及对IL-4和IFN-γ调控机制的研究
发布时间：2018-08-18 08:22
【摘要】：目的本实验在前期动物实验已确定古方天灸药物白芥子散穴位贴敷治疗哮喘最佳配比的基础上,拟从白芥子散对哮喘大鼠气道炎症的病理学改变及对免疫因子的调控角度,探究白芥子散穴位贴敷对哮喘大鼠的治疗作用,为临床治疗哮喘提供理论依据。方法以SD大鼠为研究对象,根据大鼠体重,按均衡随机法分为4组,分别为空白对照组(A组)、哮喘模型组(B组)、穴位贴敷组(C组)、布地奈德雾化组(D组),每组实验动物10只。按照实验流程对各组大鼠分别施以不同的处理方法,处死后采集标本。1.用形态学方法,测量各组大鼠主支气管与右肺中叶连接处的管腔直径,对比各组大鼠支气管管腔直径的大小,观察白芥子散对哮喘大鼠气道痉挛的影响。2.采用HE染色,通过光镜观察对比4组大鼠肺组织炎细胞浸润程度、支气管上皮细胞脱落程度、支气管管腔内黏栓分泌的多少、气道水肿及闭塞程度等,根据制定的评判标准,对相应指标进行分析统计,直观的反映白芥子散穴位贴敷对哮喘大鼠气道炎性的改善情况。3.采用免疫组化方法,观察IL-4、IFN-γ在大鼠肺组织中的分布表达情况,通过对灰度值进行分析,判断白芥子散对哮喘大鼠免疫失衡的调节。结果1.造模成功断定:与空白对照组相比,哮喘模型组大鼠在症状上及病理上有显著变化,提示造模成功。2.大鼠支气管管腔直径对比结果:通过均值对比,穴位贴敷组布地奈德组空白对照组哮喘模型组,经方差分析,P0.05,认为四组大鼠支气管管腔直径均数对比无显著差别。3.通过观察肺组织HE染色,对单项病理观察指标分析发现:哮喘模型组与空白对照组相比,P0.01,差异显著;在气道炎性浸润情况、支气管上皮细胞脱落情况及肺泡间隔增厚程度上,C组及D组与B组相比,P0.05,无差异;在大鼠支气管管腔内黏栓分泌上,B组与A组相比,P0.01,差异显著;C组与B组相比,P0.01,差异显著,D组与B组相比,P0.05,无明显差异;在大鼠终末细支气管水肿及闭塞情况上,C组及D组与B组相比,P0.05,有差异。五项病理观察指标累计总分比较,B组与A组相比,P0.01,整体差异显著,C组及D组与B组相比,P0.01,整体差异显著,C组与D组相比,P0.05,说明两种治疗方法对比无差异。4.肺组织中IL-4及IFN-γ阳性表达检测结果分析:对炎性细胞胞浆中IL-4阳性表达灰度值分析,B组与A组相比,P0.01,差异显著;C组与B组相比,P0.05,无差异;D组与B组相比,P0.05,有差异。对上皮间质细胞胞浆中IL-4阳性表达灰度值分析,B组与A组相比,P0.05,有差异;C组及D组与B组相比,P0.05,无差异。对炎性细胞胞浆中IFN-γ阳性表达灰度值分析,B组与A组相比,P0.01,差异显著;C组及D组与B组相比,P0.01,差异显著。对上皮间质细胞胞浆中IFN-γ阳性表达灰度值分析,B组与A组相比,P0.05,无差异;C组及D组与B组相比,P0.01,差异显著。结论1.本次实验所用造模方式,可引起大鼠气道炎细胞浸润、支气管上皮细胞脱落、支气管管腔内黏栓分泌增多、支气管水肿及闭塞、肺间质增宽等,且能促进大鼠肺组织中IL-4的表达,抑制IFN-γ的表达,符合哮喘的一般病理学改变,提示造模成功。2.白芥子穴位贴敷及布地奈德雾化吸入均可扩张支气管,缓解哮喘大鼠气道痉挛,但改变幅度不大。3.白芥子散穴位贴敷与布地奈德雾化吸入对哮喘大鼠病理学改变明显,治疗效果显著,且两种治疗方法对比无差异。两者均能明显减轻哮喘大鼠终末细支气管水肿及闭塞情况。在减轻哮喘大鼠支气管管腔内黏液栓分泌上,白芥子散穴位贴敷明显优于布地奈德雾化吸入。4.IL-4及IFN-γ在炎性细胞中的表达明显高于在上皮间质细胞胞浆中的表达。白芥子散穴位贴敷和布地奈德雾化吸入均能降低哮喘大鼠IL-4的表达,且能提高哮喘大鼠IFN-γ的表达。在对IL-4的调控上,布地奈德雾化吸入优于白芥子散穴位贴敷;在对IFN-γ的调控上,两者无差异。5.白芥子散穴位贴敷治疗哮喘疗效确切,与西药布地奈德雾化吸入对比,无明显差异,疗效相当。
[Abstract]:Objective Based on the best prescription of Baijiezi powder for treating asthma in animal experiment, this experiment is to explore the therapeutic effect of Baijiezi powder on asthmatic rats from the point of view of pathological changes of airway inflammation and regulation of immune factors. Methods According to the body weight of SD rats, SD rats were randomly divided into four groups: blank control group (group A), asthma model group (group B), acupoint application group (group C), budesonide atomization group (group D), 10 rats in each group. Samples were collected after death. 1. The lumen diameters at the junction of the main bronchus and the right middle lobe were measured by morphological method. The lumen diameters of bronchial tubes were compared in each group, and the effects of Baijiezi powder on airway spasm in asthmatic rats were observed. 2. The infiltration of inflammatory cells in the lung tissues of the four groups were observed by HE staining and compared by light microscope. According to the established evaluation criteria, the corresponding indicators were analyzed and statistically reflected intuitively the improvement of airway inflammation in asthmatic rats treated with Baijiezsan acupoint application. 3. Using immunohistochemical method, the levels of IL-4 and IFN-gamma were observed. Distribution and expression of pulmonary tissue in rats were analyzed by gray value to judge the regulation of Baijiezi Powder on immune imbalance in asthmatic rats. Results 1. Successful modeling concluded that: compared with the blank control group, asthmatic model group rats had significant changes in symptoms and pathology, suggesting successful modeling. 2. Contrast results of bronchial lumen diameter in rats By means of mean comparison, the acupoint application group Budesonide group blank control group asthma model group, by analysis of variance, P 0.05, that the four groups of rats bronchial lumen diameter mean contrast is not significantly different.3. By observing the lung tissue HE staining, the analysis of single pathological observation indicators found: asthma model group compared with the blank control group, P 0.01, the difference is significant. There was no significant difference between group C and group D in airway inflammatory infiltration, bronchial epithelial cell exfoliation and alveolar septal thickening compared with group B, P 0.05, P 0.01 in bronchial mucosa secretion between group B and group A, P 0.01 in group C, P 0.01 in group B, P 0.05 in group D and group B. Compared with group B, there were significant differences in the cumulative total scores of five pathological observation indexes between group B and group A. Compared with group B, there were significant differences in the cumulative total scores of five pathological observation indexes between group B and group B, and there were significant differences in group C and group D, compared with group B, P 0.01 and P 0.05. Analysis of IL-4 and IFN-gamma positive expression in tissues: The gray value of IL-4 positive expression in cytoplasm of inflammatory cells in group B was significantly different from that in group A, P 0.01; P 0.05 in group C was not different from that in group B; P 0.05 in group D was different from that in group B. There were significant differences between group C and group D and group B. There was no significant difference between group B and group C in the expression of IFN - gamma in cytoplasm of inflammatory cells. There was no significant difference between group B and group A in the expression of IFN - gamma. Compared with group B, P 0.01, the difference was significant. Conclusion 1. The modeling method used in this experiment can cause airway inflammation cells infiltration, bronchial epithelial cells exfoliation, bronchial cavity mucosal thrombus secretion increased, bronchial edema and occlusion, pulmonary interstitial widening, and can promote the expression of IL-4 in rat lung tissue, inhibit the expression of IFN-gamma, consistent with asthma. General pathological changes, suggesting the success of modeling. 2. White mustard acupoint sticking and budesonide atomization inhalation can dilate bronchi, alleviate asthmatic rats airway spasm, but the change is not significant. 3. Baijiezsan acupoint sticking and budesonide atomization inhalation on asthmatic rats with obvious pathological changes, significant therapeutic effect, and two treatment methods. Both of them can obviously relieve the edema and obstruction of the bronchioles in asthmatic rats. Baijiezsan acupoint application is superior to Budesonide aerosol inhalation in alleviating the secretion of mucus emboli in the bronchioles of asthmatic rats. 4. The expression of IL-4 and IFN-gamma in inflammatory cells is significantly higher than that in the epithelial stromal cells. Da. Baijiezi powder acupoint application and budesonide atomization inhalation can reduce the expression of IL-4 in asthmatic rats, and can increase the expression of IFN-gamma in asthmatic rats. There was no significant difference between western medicine budesonide and aerosol inhalation.
【学位授予单位】：山西中医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245
[Abstract]:Objective Based on the best prescription of Baijiezi powder for treating asthma in animal experiment, this experiment is to explore the therapeutic effect of Baijiezi powder on asthmatic rats from the point of view of pathological changes of airway inflammation and regulation of immune factors. Methods According to the body weight of SD rats, SD rats were randomly divided into four groups: blank control group (group A), asthma model group (group B), acupoint application group (group C), budesonide atomization group (group D), 10 rats in each group. Samples were collected after death. 1. The lumen diameters at the junction of the main bronchus and the right middle lobe were measured by morphological method. The lumen diameters of bronchial tubes were compared in each group, and the effects of Baijiezi powder on airway spasm in asthmatic rats were observed. 2. The infiltration of inflammatory cells in the lung tissues of the four groups were observed by HE staining and compared by light microscope. According to the established evaluation criteria, the corresponding indicators were analyzed and statistically reflected intuitively the improvement of airway inflammation in asthmatic rats treated with Baijiezsan acupoint application. 3. Using immunohistochemical method, the levels of IL-4 and IFN-gamma were observed. Distribution and expression of pulmonary tissue in rats were analyzed by gray value to judge the regulation of Baijiezi Powder on immune imbalance in asthmatic rats. Results 1. Successful modeling concluded that: compared with the blank control group, asthmatic model group rats had significant changes in symptoms and pathology, suggesting successful modeling. 2. Contrast results of bronchial lumen diameter in rats By means of mean comparison, the acupoint application group Budesonide group blank control group asthma model group, by analysis of variance, P 0.05, that the four groups of rats bronchial lumen diameter mean contrast is not significantly different.3. By observing the lung tissue HE staining, the analysis of single pathological observation indicators found: asthma model group compared with the blank control group, P 0.01, the difference is significant. There was no significant difference between group C and group D in airway inflammatory infiltration, bronchial epithelial cell exfoliation and alveolar septal thickening compared with group B, P 0.05, P 0.01 in bronchial mucosa secretion between group B and group A, P 0.01 in group C, P 0.01 in group B, P 0.05 in group D and group B. Compared with group B, there were significant differences in the cumulative total scores of five pathological observation indexes between group B and group A. Compared with group B, there were significant differences in the cumulative total scores of five pathological observation indexes between group B and group B, and there were significant differences in group C and group D, compared with group B, P 0.01 and P 0.05. Analysis of IL-4 and IFN-gamma positive expression in tissues: The gray value of IL-4 positive expression in cytoplasm of inflammatory cells in group B was significantly different from that in group A, P 0.01; P 0.05 in group C was not different from that in group B; P 0.05 in group D was different from that in group B. There were significant differences between group C and group D and group B. There was no significant difference between group B and group C in the expression of IFN - gamma in cytoplasm of inflammatory cells. There was no significant difference between group B and group A in the expression of IFN - gamma. Compared with group B, P 0.01, the difference was significant. Conclusion 1. The modeling method used in this experiment can cause airway inflammation cells infiltration, bronchial epithelial cells exfoliation, bronchial cavity mucosal thrombus secretion increased, bronchial edema and occlusion, pulmonary interstitial widening, and can promote the expression of IL-4 in rat lung tissue, inhibit the expression of IFN-gamma, consistent with asthma. General pathological changes, suggesting the success of modeling. 2. White mustard acupoint sticking and budesonide atomization inhalation can dilate bronchi, alleviate asthmatic rats airway spasm, but the change is not significant. 3. Baijiezsan acupoint sticking and budesonide atomization inhalation on asthmatic rats with obvious pathological changes, significant therapeutic effect, and two treatment methods. Both of them can obviously relieve the edema and obstruction of the bronchioles in asthmatic rats. Baijiezsan acupoint application is superior to Budesonide aerosol inhalation in alleviating the secretion of mucus emboli in the bronchioles of asthmatic rats. 4. The expression of IL-4 and IFN-gamma in inflammatory cells is significantly higher than that in the epithelial stromal cells. Da. Baijiezi powder acupoint application and budesonide atomization inhalation can reduce the expression of IL-4 in asthmatic rats, and can increase the expression of IFN-gamma in asthmatic rats. There was no significant difference between western medicine budesonide and aerosol inhalation.
【学位授予单位】：山西中医学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245
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