The experimental study on the biological rhythm expression of substance P in allergic airway inflammation mice
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摘要: 目的 建立变应性气道炎症小鼠模型，探讨P物质在变应性气道炎症小鼠中的生物节律性表达。方法 将84只BABL/c小鼠随机分为14个组，分别为实验组7组(0：00组，04：00组，08：00组，12：00组，16：00组，20：00组，24：00组)和对照组7组(0：00组，04：00组，08：00组，12：00组，16：00组，20：00组，24：00组)，每组6只。采用卵清蛋白腹腔注射+鼻腔局部激发+雾化吸入的方法制备小鼠变应性气道炎症模型。对实验动物血液进行ELISA法测量上清液P物质含量，采用RT-PCR技术检测不同时间段内P物质在鼻腔组织、肺组织及支气管组织中基因表达的分布及含量。取小鼠鼻腔组织、左肺组织及左主支气管组织行免疫组织化学染色，观察组织切片中P物质阳性细胞的表达及分布。结果 实验组各时间段内血浆中P物质含量明显高于对照组(P < 0.05)，其中，实验组中0：00组(321.99±28.93) pg/mL、04：00组(310.76±25.31) pg/mL及24：00组(335.62±28.29) pg/mL的P物质含量明显高于实验组其他时间段(P < 0.05)。实验组小鼠鼻腔组织、左肺组织及左主支气管组织中P物质mRNA表达明显高于对应的对照组(P < 0.01)。实验组小鼠鼻腔组织、左肺及左主支气管免疫组织化学切片中P物质表达显著高于对照组。结论 P物质在变应性气道炎症小鼠模型中呈现高表达，且在凌晨及上午04：00~08：00表达出现峰值，这可能为治疗变应性气道炎症的给药时间提供新的依据。Abstract: Objective To establish the murine models of allergic airway inflammation, and to investigate the biological rhythm expression of substance P in it.Method Eighty-four BALB/c mice were divided into 14 groups, which were airway inflammation groups(0: 00 group, 04: 00 group, 08: 00 group, 12: 00 group, 16: 00 group, 20: 00 group, 24: 00 group) and normal control groups(0: 00 group, 04: 00 group, 08: 00 group, 12: 00 group, 16: 00 group, 20: 00 group, 24: 00 group). The allergic airway inflammation models of mice were established by intraperitoneal injection of ovalbumin(OVA), nasal challenge and aerosol inhalation with OVA. The plasma substance P levels were measured by ELISA. The expression of substance P in nasal mucosa, left lung tissue and left bronchial tissue during different time periods was obversed with RT-PCR and immunohistochemical technique.Result In allergic airway inflammation groups, the plasma substance P levels were higher than in normal control groups(P < 0.05), and 0: 00 group(321.99±28.93) pg/mL, 04: 00 group(310.76±25.31) pg/mL and 24: 00 group(335.62±28.29) pg/mL were higher than other experimental groups(P < 0.05). The expression of substance P mRNA in nasal mucosa, left lung tissue and left bronchial tissue was higher than control groups(P < 0.01). By observing immunohistochemical sections, the expression of substance P in allergic airway inflammation groups was higer than control groups.Conclusion The expression of substance P is high in allergic airway inflammation mice and the peaks are during 04: 00-08: 00 in the early morning, which may provide new basis for treating allergic airway inflammation.
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Key words:
- allergic airway inflammation /
- substance P /
- biological rhythm
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表 1 各时间点小鼠血浆P物质含量
pg/mL，x±s 时间点 只数 实验组 对照组 t P 0：00 6 321.99±28.931)2) 166.25±11.91 12.19 < 0.01 04：00 6 310.76±25.311)2) 140.33±22.74 12.27 < 0.01 08：00 6 261.17±25.281) 111.14±8.42 13.79 < 0.01 12：00 6 238.76±23.911) 114.86±12.57 11.24 < 0.01 16：00 6 251.19±26.831) 109.33±9.42 12.22 < 0.01 20：00 6 240.91±25.911) 101.30±7.37 12.69 < 0.01 24：00 6 335.62±28.291)2) 141.04±9.40 15.99 < 0.01 与对照组比较，1)P < 0.01；与08：00、12：00、16：00、20：00比较，2)P < 0.05。 
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表 2 各时间点小鼠鼻腔组织中P物质mRNA表达
时间点 只数 实验组 对照组 t P 0：00 6 0.553±0.073 0.160±0.021 12.718 < 0.01 04：00 6 0.519±0.082 0.164±0.029 9.964 < 0.01 08：00 6 0.498±0.094 0.158±0.029 8.495 < 0.01 12：00 6 0.410±0.033 0.128±0.019 18.347 < 0.01 16：00 6 0.414±0.113 0.129±0.026 6.023 < 0.01 20：00 6 0.413±0.063 0.134±0.029 9.782 < 0.01 24：00 6 0.544±0.038 0.165±0.015 22.877 < 0.01
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表 3 各时间点小鼠左肺组织中P物质mRNA表达
时间点 只数 实验组 对照组 t P 0：00 6 0.618±0.067 0.170±0.038 14.254 < 0.01 04：00 6 0.479±0.085 0.140±0.018 9.603 < 0.01 08：00 6 0.363±0.312 0.157±0.026 12.193 < 0.01 12：00 6 0.395±0.064 0.146±0.018 9.149 < 0.01 16：00 6 0.382±0.061 0.138±0.021 9.241 < 0.01 20：00 6 0.388±0.088 0.146±0.030 6.344 < 0.01 24：00 6 0.611±0.056 0.171±0.021 18.122 < 0.01
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表 4 各时间点小鼠左主支气管组织中P物质mRNA表达
时间点 只数 实验组 对照组 t P 0：00 6 0.553±0.085 0.176±0.020 10.552 < 0.01 04：00 6 0.509±0.050 0.178±0.011 15.736 < 0.01 08：00 6 0.488±0.091 0.152±0.021 8.834 < 0.01 12：00 6 0.374±0.036 0.125±0.006 16.953 < 0.01 16：00 6 0.395±0.059 0.124±0.017 10.903 < 0.01 20：00 6 0.366±0.049 0.134±0.008 11.541 < 0.01 24：00 6 0.552±0.102 0.178±0.015 8.883 < 0.01
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