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目的检测哮喘小鼠中趋化因子受体CXCR4的表达及其拮抗剂氯喹(CQ)的干预作用。方法将6~8周SPF级Balb/c小鼠随机分成对照组、哮喘组和氯喹干预组,哮喘组和氯喹干预组用鸡卵清蛋白(OVA)致敏和激发建立哮喘小鼠模型,干预组于激发前30 min给予CQ,连续3 d,对照组用PBS代替。最后1次激发24 h内小鼠肺功能仪检测小鼠气道高反应;最后1次激发48 h内检测支气管肺泡灌洗液(BALF)中细胞总数及细胞分类计数;HE染色光镜下观察肺组织的病理炎症改变,免疫组化染色检测肺组织CXCR4的表达;荧光定量PCR(Q-PCR)测定肺组织中CXCR4 mRNA的表达。结果哮喘组气道高反应及BALF中细胞总数明显高于对照组,干预组小鼠气道高反应及BALF中细胞总数与哮喘组相比显著降低;BALF中哮喘组嗜酸性粒细胞、中性粒细胞和单核细胞数较空白组显著增加,而氯喹干预组较哮喘组显著减低;HE染色结果显示哮喘组肺部炎症细胞浸润明显,干预组与哮喘组相比明显减轻,病理评分降低;免疫组化显示哮喘组CXCR4在气道上皮细胞呈强阳性表达,干预组表达呈弱阳性;哮喘组CXCR4 mRNA的表达较对照组升高,氯喹干预组较哮喘组CXCR4 mRNA表达量降低;CXCR4表达量与BALF细胞总数呈正相关。结论 CXCR4可能参与哮喘小鼠的气道炎症和气道高反应的发生,氯喹能改善哮喘小鼠的气道高反应和气道炎症可能与拮抗CXCR4相关,为氯喹治疗哮喘提供理论和实验依据,为开发治疗哮喘新药提供新的思路。
Objective To detect the expression of chemokine receptor CXCR4 in asthmatic mice and the intervention of its antagonist chloroquine (CQ). Methods Sixty-eight weeks old SPF Balb / c mice were randomly divided into control group, asthma group and chloroquine intervention group, asthma group and chloroquine intervention group were sensitized and challenged with chicken ovalbumin (OVA) CQ was administered 30 min before challenge for 3 consecutive days and the control group was replaced by PBS. In the last one time, the airway hypersensitivity of mice was detected by pulmonary function test within 24 hours. The total number of cells and the number of cells in bronchoalveolar lavage fluid (BALF) were detected within 48 h after the last challenge. The pathological changes of lung tissue were observed. The expression of CXCR4 in lung tissue was detected by immunohistochemical staining. The expression of CXCR4 mRNA in lung tissue was detected by quantitative PCR (Q-PCR). Results The airway hyperresponsiveness and the total number of cells in BALF in asthmatic group were significantly higher than those in control group. The airway hyperresponsiveness and the total number of cells in BALF in asthmatic group were significantly decreased compared with those in asthmatic group. The levels of eosinophils, The number of granulocytes and monocytes increased significantly compared with the blank group, while the chloroquine intervention group was significantly lower than the asthma group; HE staining showed lung tissue infiltration of inflammatory cells in asthma group significantly reduced compared with the asthma group, the pathological score decreased; Immunohistochemistry showed that the expression of CXCR4 in asthmatic group was strongly positive in airway epithelial cells and the expression was weakly positive in asthmatic group. The expression of CXCR4 mRNA in asthmatic group was higher than that in control group, while the expression of CXCR4 mRNA in asthmatic group was lower than that in control group The amount of BALF cells was positively correlated. Conclusion CXCR4 may be involved in airway inflammation and airway hyperresponsiveness in asthmatic mice. Chloroquine can improve the airway hyperresponsiveness and airway inflammation in asthmatic mice and may be related to the antagonism of CXCR4, providing theoretical and experimental basis for chloroquine treatment of asthma. New treatment of asthma drugs to provide new ideas.