【摘要】： 第一部分哮喘小鼠氣道炎癥和氣道黏液分泌的改變 目的觀察哮喘小鼠氣道炎癥和氣道黏液分泌的改變方法清潔級(jí)BALB/c小鼠20只隨機(jī)分為兩組：哮喘組(AS組)10只和正常組(NS組)10只,哮喘組用卵清白蛋白(OVA)致敏和激發(fā)制作哮喘模型,正常組用生理鹽水致敏和激發(fā)。通過(guò)檢測(cè)AS組小鼠支氣管肺泡灌洗液(BALF)細(xì)胞總數(shù)和細(xì)胞分類(lèi)計(jì)數(shù),采用ELISA檢測(cè)BALF中的IL-4和TNF-α水平,通過(guò)HE染色觀察肺部病理學(xué)改變,用阿爾辛藍(lán)-過(guò)碘酸雪夫(AB-PAS)染色氣道杯狀細(xì)胞,免疫組織化學(xué)檢測(cè)肺組織中黏蛋白5ac(Muc5ac)的表達(dá)及熒光定量RT-PCR檢測(cè)Muc5acmRNA在肺內(nèi)的表達(dá)。結(jié)果哮喘組小鼠BALF中的細(xì)胞總數(shù)、嗜酸性粒細(xì)胞、單核細(xì)胞、淋巴細(xì)胞百分比,IL-4和TNF-α水平、肺組織AB-PAS陽(yáng)染面積,Muc5ac蛋白和：mRNA表達(dá)明顯高于正常對(duì)照組小鼠(P0.01或P0.05)。結(jié)論OVA致敏和激發(fā)的哮喘小鼠出現(xiàn)以嗜酸性粒細(xì)胞、淋巴細(xì)胞浸潤(rùn)為主的氣道炎癥及杯狀細(xì)胞增生的氣道黏液高分泌,且氣道炎癥和氣道黏液高分泌關(guān)系密切。 第二部分CCR3在哮喘小鼠氣道組織Muc5ac表達(dá)中的作用及SB328437干預(yù)的影響 目的通過(guò)應(yīng)用CCR3拮抗劑SB328437干預(yù)哮喘組小鼠,探討CCR3在哮喘小鼠氣道Muc5ac表達(dá)中的作用。方法清潔級(jí)BALB/c小鼠50只隨機(jī)分成正常對(duì)照組(NS組)、哮喘組(AS組)、地塞米松干預(yù)組(AS+DEX組)、SB328437干預(yù)組(AS+SB組)及溶劑對(duì)照組二甲基亞砜組(DMSO組),各10只。測(cè)定BALF中細(xì)胞總數(shù)和細(xì)胞分類(lèi)計(jì)數(shù),采用ELISA檢測(cè)BALF中的IL-4和TNF-α水平,通過(guò)HE染色觀察肺組織病理學(xué)改變,AB-PAS對(duì)氣道杯狀細(xì)胞進(jìn)行染色,用免疫組織化學(xué)法檢測(cè)氣道Muc5ac和肺組織CCR3蛋白的表達(dá)及熒光定量RT-PCR檢測(cè)Muc5ac mRNA、CCR3 mRNA在肺組織內(nèi)的表達(dá)。結(jié)果AS組在BALF細(xì)胞總數(shù)、嗜酸性粒細(xì)胞、單核細(xì)胞、淋巴細(xì)胞、IL-4,TNF-α水平、AB-PAS陽(yáng)染面積、黏蛋白Muc5ac、CCR3蛋白IOD及Muc5ac mRNA、CCR3mRNA表達(dá)與AS+SB組、AS+DEX組比較差異有統(tǒng)計(jì)學(xué)意義(P＜0.01或P0.05)。結(jié)論CCR3可能介導(dǎo)哮喘小鼠的慢性氣道炎癥反應(yīng)和導(dǎo)致氣道黏蛋白基因Muc5ac mRNA的高表達(dá)和黏蛋白Muc5ac的高分泌。SB328437可能通過(guò)抑制肺組織CCR3 mRNA及其蛋白的表達(dá)而抑制氣道炎癥和氣道黏液高分泌。
[Abstract]:Part I changes of airway inflammation and airway mucus secretion in asthmatic mice ObjectiveTo observe the changes of airway inflammation and airway mucus secretion in asthmatic mice 20 BALB/c mice of clean grade were randomly divided into two groups: Asthma group (AS group, n = 10) and normal group (NS group, n = 10), The asthmatic model was induced by ovalbumin (OVA) in asthmatic group, and the normal group was sensitized and stimulated with normal saline. The total number and cell classification of (BALF) cells in bronchoalveolar lavage fluid (BALF) of mice in AS group were measured, the levels of IL-4 and TNF- 偽 in BALF were detected by ELISA, and the pathological changes of lung were observed by HE staining. The expression of mucin 5ac (Muc5ac) in lung tissue and the expression of Muc5acmRNA in lung tissue were detected by immunohistochemistry and fluorescence quantitative RT-PCR. Results the levels of IL-4 and TNF- 偽 in BALF, eosinophil, monocyte and lymphocyte percentage in asthma group were significantly higher than those in normal control group (P0.01 or P0.05). Conclusion Airway mucus with eosinophilic granulocyte, lymphocyte infiltration and goblet cell proliferation were found in asthmatic mice sensitized and stimulated by OVA, and there was a close relationship between airway inflammation and airway mucus hypersecretion. The role of CCR3 in the expression of Muc5ac in airway tissue of asthmatic mice and the effect of SB328437 intervention objective to investigate the role of CCR3 in the expression of Muc5ac in asthmatic mice by using CCR3 antagonist SB328437. Methods Fifty clean grade BALB/c mice were randomly divided into normal control group (NS group), asthma group (AS group), dexamethasone intervention group (AS DEX group), SB328437 intervention group (AS SB group) and solvent control group (DMSO group). The total number of cells and the number of cells in BALF were measured. The levels of IL-4 and TNF- 偽 in BALF were detected by ELISA. The pathological changes of lung tissue were observed by HE staining and the goblet cells were stained by AB-PAS. Immunohistochemical method was used to detect the expression of CCR3 protein in airway and lung tissue, and fluorescence quantitative RT-PCR was used to detect the expression of Muc5ac mRNA,CCR3 mRNA in lung tissue. Results the total number of BALF cells, eosinophilic granulocyte, monocyte, lymphocyte IL-4 TNF- 偽 level and AB-PAS positive staining area, IOD and Muc5ac mRNA,CCR3mRNA expression of mucin Muc5ac,CCR3 protein in AS group were significantly different from those in AS SB group and AS SB group (P < 0. 01 or P < 0. 05). Conclusion CCR3 may mediate chronic airway inflammation in asthmatic mice and induce high expression of airway mucin gene Muc5ac mRNA and high secretion of mucin Muc5ac. SB328437 may inhibit airway inflammation by inhibiting the expression of CCR3 mRNA and its protein in lung tissue. Hypersecretion of mucus and airway mucus.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R562.25

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相關(guān)期刊論文 前1條

1 彭小華;楊遠(yuǎn);;地塞米松對(duì)哮喘小鼠氣道炎癥反應(yīng)和STAT6表達(dá)的影響[J];東南大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2009年05期

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本文編號(hào)：2224820