敲除CCR3基因对小鼠嗜酸粒细胞的作用实验观察

发布时间：2018-01-19 09:40

本文关键词： 鼻炎变应性基因敲除 CCR 小鼠嗜酸粒细胞　出处：《临床耳鼻咽喉头颈外科杂志》2017年24期 　论文类型：期刊论文

【摘要】：目的:观察CCR3基因敲除对小鼠嗜酸粒细胞(EOS)增殖、成熟及凋亡的影响。方法:利用CCR3基因敲除小鼠(实验组)及野生型小鼠(对照组),取小鼠骨髓细胞进行体外培养,诱导刺激其分化成熟为EOS,通过细胞计数观察EOS增殖情况,qRT-PCR检测脱颗粒蛋白mRNA表达量,Annexin V-FITC/PI双染法检测EOS细胞凋亡率。结果:(1)体外培养EOS第0~14天细胞计数表明,第10、12、14天实验组获得细胞数均较对照组少,增殖较慢,差异有统计学意义(P0.01);(2)细胞凋亡率检测结果表明,在含有10ng/ml IL-5或无IL-5条件下,实验组EOS细胞凋亡率均高于对照组,差异有统计学意义(P0.01);(3)实验组ECP、EPO、MBP mRNA表达量与对照组相比均有不同程度降低,差异有统计学意义(P0.05)。结论:敲除CCR3基因可抑制EOS增殖及成熟,促进其凋亡,为CCR3作为变应性鼻炎治疗靶基因提供理论依据。
[Abstract]:Objective: to observe the effect of CCR3 knockout on proliferation, maturation and apoptosis of eosinophils in mice. Methods: CCR3 gene knockout mice (experimental group) and wild-type mice (control group) were used. Mouse bone marrow cells were cultured in vitro to induce the differentiation and maturation of mouse bone marrow cells. The proliferation of EOS was observed by cell count and mRNA expression of degranulated protein was detected by qRT-PCR. Annexin V-FITC / Pi double staining method was used to detect the apoptosis rate of EOS cells. On the 14th day, the number of cells obtained in the experimental group was less than that in the control group, and the proliferation was slower. The difference was statistically significant (P 0.01). The results showed that the apoptotic rate of EOS cells in the experimental group was higher than that in the control group under the condition of 10 ng / ml IL-5 or no IL-5. The difference was statistically significant (P 0.01). (3) the expression of MBP mRNA in ECPT in the experimental group was significantly lower than that in the control group. Conclusion: knockout of CCR3 gene can inhibit the proliferation and maturation of EOS, promote its apoptosis, and provide theoretical basis for CCR3 as a target gene for the treatment of allergic rhinitis.
【作者单位】：南昌大学第二附属医院耳鼻咽喉头颈外科;
【基金】：国家自然科学基金项目(No:81360160) 江西省优势科技创新团队专项计划(No:S2016RCTDB0016)
【分类号】：R765.21
【正文快照】： 变应性鼻炎是由IgE介导的抵抗吸入性过敏原及局部黏膜炎性改变的非感染性慢性炎性疾病[1],嗜酸粒细胞(EOS)是AR炎症反应的关键性效应细胞,由骨髓多能造血干细胞发育而来,可由骨髓迁移到外周血及鼻腔局部黏膜组织中而起作用,在变应性鼻炎的发生发展过程中起着重要作用[2-4]。CC

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