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目的探讨p53在过敏性鼻炎小鼠CD4~+T细胞中的表达。方法 Balb/c小鼠随机分为三组,经霍乱毒素及卵清蛋白抗原进行基础致敏,并经卵清蛋白滴鼻激发,建立小鼠过敏性鼻炎模型并进行小鼠过敏状态评价;分离小鼠外周血单个核细胞,流式细胞仪分析CD4~+IL-13~+T细胞比率,Western blot分析外周血CD4~+T细胞中p53蛋白表达量;提取各组小鼠鼻腔粘膜RNA,实时定量PCR检测IL-13、p53、STAT6表达量。结果成功建立了过敏鼻炎小鼠模型。对照组OVA-IgE的OD值为0.006 5±0.006 2,模型组OVA-IgE为0.365 0±0.054 6;对照组组胺水平为(1 327±55.56)pg/mL,模型组组胺(1 871±136.8)pg/mL,模型组小鼠卵清蛋白特异性IgE、组胺明显增高,且存在大量Th2细胞增殖;模型组与对照组相比OVA特异性IgE及组胺表达水平增高(P<0.05);模型组PBMC中细胞增殖比率明显增加(61.2%),且CD4~+IL-4~+Th2细胞增殖比率明显增加(50.1%)。与对照组相比,模型组小鼠外周血CD4~+IL-13~+T细胞比率增加,CD4+T细胞中p53蛋白表达量下降,差异均有统计学意义(P<0.05)。对照组鼻腔粘膜中IL-13表达为0.540±0.137,模型组为1.502±0.263;对照组STAT6的RNA表达量为0.764±0.102,模型组为1.482±0.173;对照组p53蛋白RNA表达量为0.805±0.223,模型组为0.433±0.174,差异均有统计学意义(P<0.05)。鼻腔粘膜中IL-13和STAT6的RNA表达增加,而p53蛋白RNA表达下降。结论 IL-13在过敏性鼻炎小鼠外周血CD4~+T细胞及过敏性鼻腔粘膜组织中的表达增加,而p53表达量降低。
Objective To investigate the expression of p53 in CD4 ~ + T cells of allergic rhinitis mice. Methods Balb / c mice were randomly divided into three groups. The basal cells were sensitized with cholera toxin and ovalbumin antigen and challenged with ovalbumin to establish allergic rhinitis mice model. The percentage of CD4 ~ + IL-13 ~ + T cells in peripheral blood mononuclear cells was analyzed by flow cytometry and the expression of p53 protein in peripheral CD4 ~ + T cells by Western blot. Real-time quantitative PCR detection of IL-13, p53, STAT6 expression. Results The allergic rhinitis mouse model was successfully established. The OD value of OVA-IgE in the control group was 0.006 5 ± 0.006 2, while the OVA-IgE in the model group was 0.365 0 ± 0.054 6. The histamine level in the control group was (1 327 ± 55.56) pg / mL, ± 136.8) pg / mL, the specific IgE and histamine in model group were significantly increased, and there were a large number of Th2 cell proliferation. Compared with the control group, OVA-specific IgE and histamine expression increased (P < 0.05). The proliferation rate of PBMC in model group increased significantly (61.2%), and the proliferation rate of CD4 ~ + IL-4 ~ + Th2 cells increased significantly (50.1%). Compared with the control group, the percentage of CD4 ~ + IL-13 ~ + T cells in peripheral blood of model group increased and the expression of p53 protein in CD4 + T cells decreased in model group (P <0.05). The expression of IL-13 in the nasal mucosa of the control group was 0.540 ± 0.137, and the model group was 1.502 ± 0.263. The RNA expression level of STAT6 in the control group was 0.764 ± 0.102 and the model group was 1.482 ± 0.173. The expression of p53 protein in the control group was 0.805 ± 0.223, and the model group was 0.433 ± 0.174, with significant difference (P <0.05). Nasal mucosa IL-13 and STAT6 RNA expression increased, while the p53 protein RNA expression decreased. Conclusion The expression of IL-13 in CD4 ~ + T cells and allergic nasal mucosa of mice with allergic rhinitis increased, while the expression of p53 decreased.