CCR3在哮喘小鼠气道组织Muc5ac表达中的作用
发布时间：2018-09-05 16:28
【摘要】： 第一部分哮喘小鼠气道炎症和气道黏液分泌的改变 目的观察哮喘小鼠气道炎症和气道黏液分泌的改变方法清洁级BALB/c小鼠20只随机分为两组：哮喘组(AS组)10只和正常组(NS组)10只,哮喘组用卵清白蛋白(OVA)致敏和激发制作哮喘模型,正常组用生理盐水致敏和激发。通过检测AS组小鼠支气管肺泡灌洗液(BALF)细胞总数和细胞分类计数,采用ELISA检测BALF中的IL-4和TNF-α水平,通过HE染色观察肺部病理学改变,用阿尔辛蓝-过碘酸雪夫(AB-PAS)染色气道杯状细胞,免疫组织化学检测肺组织中黏蛋白5ac(Muc5ac)的表达及荧光定量RT-PCR检测Muc5acmRNA在肺内的表达。结果哮喘组小鼠BALF中的细胞总数、嗜酸性粒细胞、单核细胞、淋巴细胞百分比,IL-4和TNF-α水平、肺组织AB-PAS阳染面积,Muc5ac蛋白和：mRNA表达明显高于正常对照组小鼠(P0.01或P0.05)。结论OVA致敏和激发的哮喘小鼠出现以嗜酸性粒细胞、淋巴细胞浸润为主的气道炎症及杯状细胞增生的气道黏液高分泌,且气道炎症和气道黏液高分泌关系密切。 第二部分CCR3在哮喘小鼠气道组织Muc5ac表达中的作用及SB328437干预的影响 目的通过应用CCR3拮抗剂SB328437干预哮喘组小鼠,探讨CCR3在哮喘小鼠气道Muc5ac表达中的作用。方法清洁级BALB/c小鼠50只随机分成正常对照组(NS组)、哮喘组(AS组)、地塞米松干预组(AS+DEX组)、SB328437干预组(AS+SB组)及溶剂对照组二甲基亚砜组(DMSO组),各10只。测定BALF中细胞总数和细胞分类计数,采用ELISA检测BALF中的IL-4和TNF-α水平,通过HE染色观察肺组织病理学改变,AB-PAS对气道杯状细胞进行染色,用免疫组织化学法检测气道Muc5ac和肺组织CCR3蛋白的表达及荧光定量RT-PCR检测Muc5ac mRNA、CCR3 mRNA在肺组织内的表达。结果AS组在BALF细胞总数、嗜酸性粒细胞、单核细胞、淋巴细胞、IL-4,TNF-α水平、AB-PAS阳染面积、黏蛋白Muc5ac、CCR3蛋白IOD及Muc5ac mRNA、CCR3mRNA表达与AS+SB组、AS+DEX组比较差异有统计学意义(P＜0.01或P0.05)。结论CCR3可能介导哮喘小鼠的慢性气道炎症反应和导致气道黏蛋白基因Muc5ac mRNA的高表达和黏蛋白Muc5ac的高分泌。SB328437可能通过抑制肺组织CCR3 mRNA及其蛋白的表达而抑制气道炎症和气道黏液高分泌。
[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- 伪 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- 伪 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- 伪 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- 伪 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.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R562.25
本文编号：2224820
[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- 伪 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- 伪 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- 伪 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- 伪 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.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R562.25
【参考文献】
相关期刊论文 前1条
1 彭小华;杨远;;地塞米松对哮喘小鼠气道炎症反应和STAT6表达的影响[J];东南大学学报(医学版);2009年05期
，本文编号：2224820
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