CCR3在哮喘小鼠氣道組織Muc5ac表達中的作用
發(fā)布時間：2018-09-05 16:28
【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BALB/c小鼠20只隨機分為兩組：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)。通過檢測AS組小鼠支氣管肺泡灌洗液(BALF)細胞總數和細胞分類計數,采用ELISA檢測BALF中的IL-4和TNF-α水平,通過HE染色觀察肺部病理學改變,用阿爾辛藍-過碘酸雪夫(AB-PAS)染色氣道杯狀細胞,免疫組織化學檢測肺組織中黏蛋白5ac(Muc5ac)的表達及熒光定量RT-PCR檢測Muc5acmRNA在肺內的表達。結果哮喘組小鼠BALF中的細胞總數、嗜酸性粒細胞、單核細胞、淋巴細胞百分比,IL-4和TNF-α水平、肺組織AB-PAS陽染面積,Muc5ac蛋白和：mRNA表達明顯高于正常對照組小鼠(P0.01或P0.05)。結論OVA致敏和激發(fā)的哮喘小鼠出現以嗜酸性粒細胞、淋巴細胞浸潤為主的氣道炎癥及杯狀細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系密切。 第二部分CCR3在哮喘小鼠氣道組織Muc5ac表達中的作用及SB328437干預的影響 目的通過應用CCR3拮抗劑SB328437干預哮喘組小鼠,探討CCR3在哮喘小鼠氣道Muc5ac表達中的作用。方法清潔級BALB/c小鼠50只隨機分成正常對照組(NS組)、哮喘組(AS組)、地塞米松干預組(AS+DEX組)、SB328437干預組(AS+SB組)及溶劑對照組二甲基亞砜組(DMSO組),各10只。測定BALF中細胞總數和細胞分類計數,采用ELISA檢測BALF中的IL-4和TNF-α水平,通過HE染色觀察肺組織病理學改變,AB-PAS對氣道杯狀細胞進行染色,用免疫組織化學法檢測氣道Muc5ac和肺組織CCR3蛋白的表達及熒光定量RT-PCR檢測Muc5ac mRNA、CCR3 mRNA在肺組織內的表達。結果AS組在BALF細胞總數、嗜酸性粒細胞、單核細胞、淋巴細胞、IL-4,TNF-α水平、AB-PAS陽染面積、黏蛋白Muc5ac、CCR3蛋白IOD及Muc5ac mRNA、CCR3mRNA表達與AS+SB組、AS+DEX組比較差異有統(tǒng)計學意義(P＜0.01或P0.05)。結論CCR3可能介導哮喘小鼠的慢性氣道炎癥反應和導致氣道黏蛋白基因Muc5ac mRNA的高表達和黏蛋白Muc5ac的高分泌。SB328437可能通過抑制肺組織CCR3 mRNA及其蛋白的表達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.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R562.25
本文編號：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- 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.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R562.25
【參考文獻】
相關期刊論文 前1條
1 彭小華;楊遠;;地塞米松對哮喘小鼠氣道炎癥反應和STAT6表達的影響[J];東南大學學報(醫(yī)學版);2009年05期
，本文編號：2224820
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