文章摘要
王 硕,王真奎,张婉莹,李艳丽,薛 丽.小鼠哮喘发作过程中相关炎症因子的动态变化[J].,2019,19(18):3428-3431
小鼠哮喘发作过程中相关炎症因子的动态变化
The Time Course Change of Cytokines in Asthmatic Mice Model
投稿时间:2019-02-28  修订日期:2019-03-23
DOI:10.13241/j.cnki.pmb.2019.18.006
中文关键词: 支气管哮喘  白介素 13  嗜酸性粒细胞趋化因子  单核细胞趋化因子  肿瘤坏死因子α
英文关键词: Bronchial asthma  Interleukin-13  Eotaxin  MCP-1  TNF-α
基金项目:黑龙江省卫生厅科研项目(2012-718)
作者单位E-mail
王 硕 哈尔滨医科大学附属第四医院 检验科 黑龙江 哈尔滨 150001 wangshuo0808@sina.com 
王真奎 哈尔滨医科大学附属第一医院 神经内科 黑龙江 哈尔滨 150001  
张婉莹 哈尔滨医科大学附属第四医院 检验科 黑龙江 哈尔滨 150001  
李艳丽 黑龙江省农垦总局总医院 检验科 黑龙江 哈尔滨 150001  
薛 丽 哈尔滨医科大学附属第四医院 检验科 黑龙江 哈尔滨 150001  
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中文摘要:
      摘要 目的:研究实验性哮喘小鼠模型在诱导哮喘发作不同时间点外周血中细胞因子IL-13、Eotaxin、MCP-1和TNF-α以及肺部浸润的炎症细胞数量变化。方法:将25只健康6-8周龄的雌性BALB/c小鼠随机分成模型组和对照组。利用卵清蛋白(OVA)诱导建立小鼠哮喘模型,模型组(Asthma)小鼠于第0、7天经腹腔注射卵清蛋白(OVA)致敏小鼠。第14-20天连续7天用1%OVA雾化激发小鼠哮喘发作,每次20 min,观察临床症状。正常对照组(Control) 小鼠以0.9%NaCL代替0VA进行腹腔注射和雾化吸入。比较两组小鼠肺组织病理切片HE染色结果、肺泡灌洗液(BALF)中炎性细胞分类计数及细胞因子IL-13、Eotaxin、MCP-1和TNF-α的浓度变化。结果:模型组小鼠BALF中IL-13的水平在试验早期(致敏2天)即开始上升达68.9±4.34,此时Eotaxin和MCP-1未见明显升高;致敏7天时IL-13、Eotaxin 和MCP-1均明显高于对照组,分别为88.3±3.39、67.4±4.24和38.9±3.1;激发1天组小鼠BALF中IL-13、Eotaxin 和MCP-1浓度持续增高至最后一次激发后1天;而TNF-α在激发1天时出现明显升高达136.9±11.9,持续到最后一次激发后1天;从激发1天肺泡灌洗液染色显微镜下观察明以淋巴细胞和嗜酸性粒细胞浸润为主。肺组织HE染色显示哮喘组小鼠气道上皮有不同程度脱落,支气管平滑肌显著增厚,血管周围水肿,炎性细胞浸润。结论:在哮喘发生过程中,IL-13水平在致敏初期即开始升高,随着继续给予OVA,Eotaxin 和MCP-1水平呈现显著增高;并伴随越来越多炎症细胞在肺部浸润,TNF-α水平出现缓慢增高,进而加重哮喘发作。
英文摘要:
      ABSTRACT Objective: To investigate the variation of IL-13, Eotaxin、MCP-1 and TNF-α in BALF from sensitized to challenge of asthmatic mice, and further explored the inflammatory cell infiltration in lung tissue. Methods: The model of asthma in mice was established by ovalbumin (OVA) sensitizing 25 BALB/c mice were randomly divided into five groups included control group, sensitized on 2 days and 7 days, challenged on 1 day, last challenged after 1 day. Each group contains five mice. All sensitized and challenged groups were administered intraperitoneally 0.1 mL of 0.01% OVA and 0.1 mL of 2% Al(OH)3. The bronchoalveolar lavage fluid (BALF) levels of IL-13, Eotaxin, MCP-1 and TNF-α were assessed by enzyme-linked immunosorbent assay (ELISA); The differential cell count was prepared using a cytospin and stained with Wright-Giemsa during OVA sensitize and challenge. The pathocytological analysis was performed in the lung tissue with HE staining. Results: The level of IL-13 was increased at sensitized on days 2 (P<0.05) for 68.9±4.34 in BALF. From sensitized on days 7 the levels of IL-13, Eotaxin and MCP-1 were significantly higher than the control group,were 88.3±3.39, 67.4±4.24 and 38.9±3.1 respectively. From challenged on 1 day the levels of IL-13, Eotaxin, and MCP-1 persist to increase until last challenged after 1 days (P<0.001). TNF-α was increased from challenged on 1 day, was 136.9±11.9. It persisted to after last challenged 1 days (P<0.05). At challenged on 1 day the numbers of lymphocytes and eosinophils were increased (P<0.05) and the number of inflammatory cells was significantly increased. Conclusion: Our studies suggest that at initial stage of OVA sensitize IL-13 up-regulated to promote secretion of Eotaxin and MCP-1 and inflammatory cells infiltration, These cytokines play important roles in asthma which improve study the pathogenesis of asthma.
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