P物质在变应性气道炎症小鼠中的生物节律性表达

The experimental study on the biological rhythm expression of substance P in allergic airway inflammation mice

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。

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 14groups,which were airway inflammation groups(0:00group,04:00group,08:00group,12:00group,16:00group,20:00group,24:00group)and normal control groups(0:00group,04:00group,08:00group,12:00group,16:00group,20:00group,24:00group).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 0:00group(321.99±28.93)pg/mL,04:00group(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 04:00-08:00in the early morning,which may provide new basis for treating allergic airway inflammation.