干扰RNA沉默基因CCR3对小鼠变应性鼻炎嗜酸性粒细胞的作用研究

发布时间：2018-06-28 16:21

本文选题：变态反应性鼻炎 + 嗜酸性粒细胞　；参考：《南昌大学》2012年硕士论文

【摘要】：目的：采用体外合成的干扰RNA，特异性抑制嗜酸性粒细胞上CCR3表达，阻断Eotaxin/CCR3路径的作用，体内观察变应性鼻炎小鼠骨髓、外周血及鼻腔灌洗液中嗜酸性粒细胞CCR3及颗粒蛋白的改变，以期了解在此过程中对嗜酸性细胞的作用及影响程度，为变应性鼻炎寻找新的治疗方法。方法：采用鼻腔局部给药，将体外合成的特异性CCR3siRNA重组穿梭质粒（pLVX-ShRNA2-mCCR3-1+2+3+4）应用于小鼠过敏性鼻炎模型上，应用逆转录酶-聚合酶链反应（RT-PCR）的方法分别检测小鼠骨髓、外周血及鼻腔灌洗液中嗜酸性粒细胞CCR3及颗粒蛋白mRNA的表达水平。结果：①在骨髓、外周血、鼻腔灌洗液中，CCR3siRNA治疗组的CCR3mRNA表达低于PBS治疗对照及siRNA治疗对照组（P0.05）；②在骨髓、外周血、鼻腔灌洗液中，CCR3siRNA治疗组的嗜酸性粒细胞颗粒蛋白--主要碱性蛋白（MBP）、阳离子蛋白（ECP）、过氧化物酶（EPO）的mRNA表达均低于PBS治疗对照及siRNA治疗对照组（P0.05）。结论：在获得高效、特异性干涉序列基础上，针对基因CCR3，给予RNAi技术的靶向治疗能够抑制变应性鼻炎动物模型中嗜酸性粒细胞的发育、迁移及浸润过程，从而减少嗜酸性粒细胞脱颗粒作用，，从而达到减轻变应性鼻炎的炎症反应。
[Abstract]:Objective: to specifically inhibit the expression of CCR3 on eosinophils, block the eotaxin / CCR3 pathway and observe the bone marrow of allergic rhinitis mice by interference RNAs in vitro. The changes of eosinophilic granulocyte CCR3 and granulocyte in peripheral blood and nasal lavage fluid in order to understand the role and influence of eosinophil in this process and to find a new treatment for allergic rhinitis. Methods: the specific CCR3siRNA recombinant shuttle plasmid (pLVX-ShRNA2-mCCR3-1234) was used in the mouse allergic rhinitis model. The mouse bone marrow was detected by reverse transcriptase polymerase chain reaction (RT-PCR). Expression of eosinophil CCR3 and granule protein mRNA in peripheral blood and nasal lavage fluid. Results the expression of CCR3 mRNA in bone marrow, peripheral blood and nasal lavage fluid was significantly lower in CCR3siRNA group than that in PBS control group and siRNA treatment control group (P0.05). The mRNA expressions of eosinophil granulocyte protein (MBP), cationic protein (ECP) and peroxidase (EPO) in nasal lavage fluid of CCR3 siRNA treatment group were lower than those of PBS control group and siRNA treatment control group (P0.05). Conclusion: on the basis of obtaining efficient and specific interference sequences, targeted therapy with RNAi for CCR3 gene can inhibit the development, migration and infiltration of eosinophils in allergic rhinitis animal model. Thus, the eosinophil degranulation can be reduced, and the inflammatory response of allergic rhinitis can be alleviated.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R765.21

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