中性粒细胞为主型尘螨过敏哮喘小鼠模型的建立

Establishment of a mouse neutrophil-dominated house dust mite allergic asthma model

目的使用屋尘螨提取物(HDM)建立中性粒细胞为主型小鼠哮喘模型。方法按照随机数字表法,将16只无特定病原体(SPF)级BALB/c小鼠分为对照组和模型组。模型组给予HDM滴鼻致敏及激发,对照组小鼠给予等量生理盐水。末次激发后24 h,采用BUXCO小鼠无创肺功能仪测量气道呼气间歇(Penh)值确定气道阻力,收集支气管肺泡灌洗液(BALF)计数细胞数并进行炎细胞分类,HE染色观察肺组织炎细胞浸润程度。实时定量PCR检测白细胞介素6(IL-6)、IL-17F、IL-1βmRNA表达,微量样本多指标流式蛋白定量技术(CBA)检测BALF中IL-4、IL-6、IL-17F、肿瘤坏死因子α(TNF-α)水平。结果与对照组相比,模型组Penh值明显升高,气管及血管旁炎症比对照组明显加重;模型组小鼠IL-6、IL-17F、IL-1βmRNA水平增加,BALF细胞数及中性粒细胞数明显增加,BALF中促中性粒细胞型细胞因子IL-6、IL-17F、TNF-α水平明显升高。结论成功建立了中性粒细胞为主型尘螨过敏小鼠哮喘模型。

Objective To establish a mouse neutrophil-dominated asthma model using house dust mite(HDM).Methods With a random number table,a total of 16 specific pathogen-free(SPF)BALB/c mice were divided into control group and model group.The model group was sensitized and stimulated by HDM nasal drip,and the control group mice were given the same amount of saline.In 24 hours after the last stimulation,the enhanced pause(Penh)was measured by the BUXCO noninvasive lung function detector to determine the airway resistance;the bronchoalveolar lavage fluid(BALF)was collected to count cells and classify inflammatory cells.The infiltration degree of inflammatory cells in the lung tissue was observed by HE staining.The expression of IL-6,IL-17F and IL-1βmRNA was detected by real-time quantitative PCR,and the levels of IL-4,IL-6,IL-17F and tumor necrosis factorα(TNF-α)in BALF were detected by cytometric bead array(CBA).Results Compared with the control group,the value of Penh in the model group was significantly higher;the tracheal and paravascular inflammation significantly aggravated;the levels of IL-6,IL-17F,IL-1βmRNA,the cell count and neutrophil count in BALF,and the levels of neutrophil cytokines IL-6,IL-17F and TNF-αin the BALF increased significantly.Conclusion The mouse neutrophil-dominated house dust mite allergic asthma model has been successfully established.