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(P<0.01);(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(P<0.01);(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(P<0.05)。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。
Objective：To investigate the effect of CCR3 gene knockout on the proliferation,maturation and apoptosis of eosinophils（EOS）in mice.Method：Bone marrow cells from CCR3 gene knockout mice（experimental group）and wild-type mice（control group）were cultured in vitro and induced differentiation into mature EOS.EOS proliferation was observed by cell counting.Expression of degranulation protein mRNA was detected by qRTPCR.EOS apoptosis was detected by Annexin V-FITC/PI double staining method.Result：（1）The number of cells on the 0-14 day of EOS cultured in vitro showed that the number of cells in the experimental group was less than that in the control group at tenth,twelfth,fourteenth days,and the proliferation was slower,the difference was statistically significant（P0.01）.（2）The test results of EOS cells apoptosis showed that,under the condition of containing 10 ng/ml IL-5 or no IL-5,the apoptosis rate of EOS cells in experimental group was higher than that in normal control group,the difference between both groups was statistically significant（P0.01）.（3）The expression results of ECP,EPO,MBP mRNA in EOS showed that the expression levels of ECP,EPO,MBP mRNA in the experimental group compared with the normal control group were reduced in varying degrees,the difference between both groups was statistically significant（P0.05）.Conclusion：Knockout CCR3 gene can inhibit the proliferation and maturation of EOS and promote its apoptosis,which provides a theoretical basis for CCR3 as a target gene to treat allergic rhinitis.
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
北大核心
2017年第24期1913-1918,共6页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
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)
