【摘要】：目的：觀察哮喘小鼠氣道炎癥及氣道黏液分泌情況。 方法：清潔級BALB/c小鼠20只隨機分為2組：哮喘組(AS組)10只,生理鹽水對照組(NS組)10只。哮喘組以卵清白蛋白(OVA)致敏及激發(fā)制作哮喘模型,生理鹽水對照組以生理鹽水代替OVA,余處理同哮喘組。支氣管肺泡灌洗液(BALF)用作細胞總數(shù)及細胞分類計數(shù),ELISA檢測支氣管肺泡灌洗液中IL-5及IL-32水平,HE染色觀察小鼠氣道及肺組織病理變化,阿爾辛藍-過碘酸雪夫染色(AB-PAS)觀察氣道組織杯狀細胞、黏液分泌情況,免疫組織化學法(IHC)檢測黏蛋白5AC(MUC5AC)在氣道及肺組織分布情況,實時熒光定量PCR、蛋白免疫印跡法(Western blot)分別檢測MUC5AC mRNA、MUC5AC蛋白在氣道組織表達。 結(jié)果：AS組小鼠BALF細胞總數(shù)、中性粒細胞數(shù)、淋巴細胞及嗜酸性粒細胞水平均高于NS組(P0.05),BALF中IL-5、IL-32水平、氣道組織AB-PAS陽性染色面積、MUC5AC蛋白及mRNA表達均高于NS組(P0.05)。 結(jié)論：哮喘小鼠存在氣道炎癥、氣道黏液高分泌。 目的：觀察辛伐他汀對哮喘小鼠氣道黏液高分泌的影響,探討辛伐他汀在哮喘小鼠氣道組織MUC5AC表達中作用機制。 方法：將清潔級BALB/c小鼠50只隨機分成5組：生理鹽水對照組(NS組)、哮喘組(AS組)、辛伐他汀干預組(SIM組)、溶劑對照組(ET組)、地塞米松組(DEX組),每組10只。進行BALF細胞總數(shù)、白細胞分類計數(shù),ELISA檢測BALF中IL-5、IL-32水平,HE染色觀察小鼠肺及氣道組織病理變化,AB-PAS染色觀察氣道組織杯狀細胞、黏液分泌情況,IHC、Western blot、實時熒光定量PCR檢測MUC5AC蛋白及mRNA在氣道及肺組織的表達。 結(jié)果：AS組BALF細胞總數(shù)、中性粒細胞、淋巴細胞及嗜酸性粒細胞數(shù)、IL-5及IL-32水平、AB-PAS陽性染色面積、MUC5AC mRNA及蛋白水平、MUC5AC積分光密度值均較NS組、SIM組、DEX組增高,差異有統(tǒng)計學意義(P0.05)；與ET組相比,差異無統(tǒng)計學意義(P0.05)；DEX組、SIM組間差異無統(tǒng)計學意義(P0.05)。 結(jié)論：辛伐他汀可抑制哮喘氣道炎癥,并且降低氣道粘蛋白MUC5AC高分泌。
[Abstract]:Objective: to observe airway inflammation and airway mucus secretion in asthmatic mice. Methods: twenty clean grade BALB/c mice were randomly divided into two groups: asthma group (AS group, n = 10) and saline control group (NS group, n = 10). The asthmatic model was induced by ovalbumin (OVA) in asthmatic group. The control group was treated with saline instead of OVA,. Bronchoalveolar lavage fluid (BALF) was used as the count of total cells and cell classification. The levels of IL-5 and IL-32 in bronchoalveolar lavage fluid (BALF) were detected by Elisa and HE staining was used to observe the pathological changes of airway and lung tissue in mice. The goblet cells and mucus secretion of airway tissue were observed by AB-PAS, and the distribution of mucin 5AC (MUC5AC) in airway and lung tissue was detected by immunohistochemical (IHC). The expression of MUC5AC mRNA,MUC5AC protein in airway tissue was detected by real-time quantitative PCR, Western blot (Western blot). Results the total number of BALF cells, the number of neutrophils, the level of lymphocyte and eosinophil in as group were higher than those in NS group (P0.05), and the expression of MUC5AC protein and mRNA in AB-PAS positive staining area of airway tissue were higher than those in NS group (P0.05). Conclusion: asthma mice have airway inflammation and airway mucus secretion. Aim: to observe the effect of simvastatin on airway mucus hypersecretion in asthmatic mice and to explore the mechanism of simvastatin on the expression of MUC5AC in airway tissue of asthmatic mice. Methods: fifty BALB/c mice of clean grade were randomly divided into 5 groups: normal saline control group (NS group), asthma group (AS group), simvastatin intervention group (SIM group), solvent control group (ET group) and dexamethasone group (DEX group) with 10 rats in each group. The total number of BALF cells and the level of IL-5,IL-32 in BALF were detected by Elisa. The pathological changes of lung and airway tissues in mice were observed by HE staining. The goblet cells in airway tissue were observed by AB-PAS staining. The expression of MUC5AC protein and mRNA in airway and lung tissues were detected by real-time fluorescence quantitative PCR (PCR). Results the total number of BALF cells, the number of neutrophils, lymphocytes and eosinophilic granulocytes and the levels of AB-PAS positive staining area of MUC5AC mRNA and protein in as group were significantly higher than those in NS group (P0.05). Compared with the ET group, the difference was not statistically significant (P0.05). Conclusion: simvastatin can inhibit airway inflammation and reduce airway mucin MUC5AC hypersecretion.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R562.25

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