Zhu X H, Wang J L, Huang Q L, Jiang Y L, Peng H S, Wu S H, Liu Y H
Department of Otorhinolaryngology Head and Neck Surgery, Second Affiliated Hospital of Nanchang University, Nanchang, 330006, China.
Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2017 Dec 20;31(24):1913-1918. doi: 10.13201/j.issn.1001-1781.2017.24.011.
To investigate the effect of CCR3 gene knockout on the proliferation, maturation and apoptosis of eosinophils (EOS) in mice. 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 qRT-PCR. EOS apoptosis was detected by Annexin V-FITC/PI double staining method. ①The 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 (<0.01). ②The 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 (<0.01). ③The 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 (<0.05). 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.
探讨CCR3基因敲除对小鼠嗜酸性粒细胞（EOS）增殖、成熟及凋亡的影响。将CCR3基因敲除小鼠（实验组）和野生型小鼠（对照组）的骨髓细胞进行体外培养，诱导分化为成熟EOS。通过细胞计数观察EOS增殖情况。采用qRT-PCR检测脱颗粒蛋白mRNA表达。用Annexin V-FITC/PI双染法检测EOS凋亡情况。①体外培养EOS 0至14天的细胞数量显示，实验组在第10天、第12天、第14天的细胞数量少于对照组，增殖较慢，差异有统计学意义（<0.01）。②EOS细胞凋亡检测结果显示，在含10 ng/ml IL-5或无IL-5条件下，实验组EOS细胞凋亡率高于正常对照组，两组差异有统计学意义（<0.01）。③EOS中ECP、EPO、MBP mRNA表达结果显示，实验组与正常对照组相比，ECP、EPO、MBP mRNA表达水平均有不同程度降低，两组差异有统计学意义（<0.05）。敲除CCR3基因可抑制EOS增殖和成熟并促进其凋亡，为CCR3作为治疗变应性鼻炎的靶基因提供了理论依据。
