【摘要】：目的本實驗在前期動物實驗已確定古方天灸藥物白芥子散穴位貼敷治療哮喘最佳配比的基礎(chǔ)上,擬從白芥子散對哮喘大鼠氣道炎癥的病理學(xué)改變及對免疫因子的調(diào)控角度,探究白芥子散穴位貼敷對哮喘大鼠的治療作用,為臨床治療哮喘提供理論依據(jù)。方法以SD大鼠為研究對象,根據(jù)大鼠體重,按均衡隨機法分為4組,分別為空白對照組(A組)、哮喘模型組(B組)、穴位貼敷組(C組)、布地奈德霧化組(D組),每組實驗動物10只。按照實驗流程對各組大鼠分別施以不同的處理方法,處死后采集標(biāo)本。1.用形態(tài)學(xué)方法,測量各組大鼠主支氣管與右肺中葉連接處的管腔直徑,對比各組大鼠支氣管管腔直徑的大小,觀察白芥子散對哮喘大鼠氣道痙攣的影響。2.采用HE染色,通過光鏡觀察對比4組大鼠肺組織炎細(xì)胞浸潤程度、支氣管上皮細(xì)胞脫落程度、支氣管管腔內(nèi)黏栓分泌的多少、氣道水腫及閉塞程度等,根據(jù)制定的評判標(biāo)準(zhǔn),對相應(yīng)指標(biāo)進(jìn)行分析統(tǒng)計,直觀的反映白芥子散穴位貼敷對哮喘大鼠氣道炎性的改善情況。3.采用免疫組化方法,觀察IL-4、IFN-γ在大鼠肺組織中的分布表達(dá)情況,通過對灰度值進(jìn)行分析,判斷白芥子散對哮喘大鼠免疫失衡的調(diào)節(jié)。結(jié)果1.造模成功斷定:與空白對照組相比,哮喘模型組大鼠在癥狀上及病理上有顯著變化,提示造模成功。2.大鼠支氣管管腔直徑對比結(jié)果:通過均值對比,穴位貼敷組布地奈德組空白對照組哮喘模型組,經(jīng)方差分析,P0.05,認(rèn)為四組大鼠支氣管管腔直徑均數(shù)對比無顯著差別。3.通過觀察肺組織HE染色,對單項病理觀察指標(biāo)分析發(fā)現(xiàn):哮喘模型組與空白對照組相比,P0.01,差異顯著;在氣道炎性浸潤情況、支氣管上皮細(xì)胞脫落情況及肺泡間隔增厚程度上,C組及D組與B組相比,P0.05,無差異;在大鼠支氣管管腔內(nèi)黏栓分泌上,B組與A組相比,P0.01,差異顯著;C組與B組相比,P0.01,差異顯著,D組與B組相比,P0.05,無明顯差異;在大鼠終末細(xì)支氣管水腫及閉塞情況上,C組及D組與B組相比,P0.05,有差異。五項病理觀察指標(biāo)累計總分比較,B組與A組相比,P0.01,整體差異顯著,C組及D組與B組相比,P0.01,整體差異顯著,C組與D組相比,P0.05,說明兩種治療方法對比無差異。4.肺組織中IL-4及IFN-γ陽性表達(dá)檢測結(jié)果分析:對炎性細(xì)胞胞漿中IL-4陽性表達(dá)灰度值分析,B組與A組相比,P0.01,差異顯著;C組與B組相比,P0.05,無差異;D組與B組相比,P0.05,有差異。對上皮間質(zhì)細(xì)胞胞漿中IL-4陽性表達(dá)灰度值分析,B組與A組相比,P0.05,有差異;C組及D組與B組相比,P0.05,無差異。對炎性細(xì)胞胞漿中IFN-γ陽性表達(dá)灰度值分析,B組與A組相比,P0.01,差異顯著;C組及D組與B組相比,P0.01,差異顯著。對上皮間質(zhì)細(xì)胞胞漿中IFN-γ陽性表達(dá)灰度值分析,B組與A組相比,P0.05,無差異;C組及D組與B組相比,P0.01,差異顯著。結(jié)論1.本次實驗所用造模方式,可引起大鼠氣道炎細(xì)胞浸潤、支氣管上皮細(xì)胞脫落、支氣管管腔內(nèi)黏栓分泌增多、支氣管水腫及閉塞、肺間質(zhì)增寬等,且能促進(jìn)大鼠肺組織中IL-4的表達(dá),抑制IFN-γ的表達(dá),符合哮喘的一般病理學(xué)改變,提示造模成功。2.白芥子穴位貼敷及布地奈德霧化吸入均可擴張支氣管,緩解哮喘大鼠氣道痙攣,但改變幅度不大。3.白芥子散穴位貼敷與布地奈德霧化吸入對哮喘大鼠病理學(xué)改變明顯,治療效果顯著,且兩種治療方法對比無差異。兩者均能明顯減輕哮喘大鼠終末細(xì)支氣管水腫及閉塞情況。在減輕哮喘大鼠支氣管管腔內(nèi)黏液栓分泌上,白芥子散穴位貼敷明顯優(yōu)于布地奈德霧化吸入。4.IL-4及IFN-γ在炎性細(xì)胞中的表達(dá)明顯高于在上皮間質(zhì)細(xì)胞胞漿中的表達(dá)。白芥子散穴位貼敷和布地奈德霧化吸入均能降低哮喘大鼠IL-4的表達(dá),且能提高哮喘大鼠IFN-γ的表達(dá)。在對IL-4的調(diào)控上,布地奈德霧化吸入優(yōu)于白芥子散穴位貼敷;在對IFN-γ的調(diào)控上,兩者無差異。5.白芥子散穴位貼敷治療哮喘療效確切,與西藥布地奈德霧化吸入對比,無明顯差異,療效相當(dāng)。
[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.
【學(xué)位授予單位】：山西中醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R245

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本文編號：2188916