RSV感染影响裸鼠气道炎症和气道高反应性的作用与机制研究
发布时间：2018-09-14 20:31
【摘要】：目的：建立呼吸道合胞病毒(RSV)感染裸鼠模型,探讨T细胞缺陷对RSV感染所致气道炎症和气道高反应性的影响。 方法：SPF级6-8周雌性裸鼠及BALB/c鼠随机分为对照组和RSV感染组,在10%水合氯醛3μl/g麻醉下,予以1.5×108PFU/ml RSV或DMEM培养基100μl滴鼻后第3、5、7及14天评估气道高反应性,并检测支气管肺泡灌洗液(BALF)中细胞总数和白细胞分类计数,HE染色检测肺部炎症病理,ELSIA法检测肺泡灌洗液中细胞因子水平。并在第5天Q-PCR检测裸鼠胆碱能受体M2(M2R)及Toll样受体3(TLR3)的mRNA表达。 结果：RSV感染后在裸鼠肺部复制持续时间更长,病毒滴度更高。裸鼠RSV感染组和BALB/c RSV感染组支气管肺泡灌洗液中细胞总数、肺部炎症病理评分均明显高于对照组(P0.05)。支气管肺泡灌洗液中淋巴细胞比例增加,巨噬细胞比例减低；HE染色显示肺部炎症以淋巴细胞浸润,血管壁、支气管壁及肺泡间隔增厚为主,其中BALB/cRSV感染后炎症较裸鼠RSV感染持续更久。裸鼠及BALB/c鼠肺功能检测显示感染后气道反应性均较其对照组增加,以BALB/c鼠RSV感染后第5天气道反应性最高,与同期裸鼠RSV感染组比较有统计学差异(P0.05)。细胞因子IFN-γ在BALB/c RSV感染后明显增加,而在裸鼠RSV感染后与其对照组比较没有显著改变。裸鼠RSV感染后TLR3mRNA均明显高于对照组(P0.05),而IL-4、IL-10等细胞因子以及胆碱能受体M2与对照组比较无显著差异(P0.05)。 结论：裸鼠感染RSV后病毒复制持续时间更长,但肺部炎症持续时间较BALB/c鼠短,气道反应性显著低于BALB/c RSV感染组,BALF中IFN-γ水平明显低于BALB/c鼠,提示RSV感染后气道炎症与气道高反应性不完全平行,T细胞及其分泌的细胞因子(如[FN-γ等)可能在RSV感染后气道病理变化中起重要作用。
[Abstract]:Aim: to establish a nude mouse model of respiratory syncytial virus (RSV) (RSV) infection and to investigate the effect of T cell deficiency on airway inflammation and airway hyperresponsiveness induced by RSV infection. Methods female nude mice and BALB/c mice of 6-8 weeks were randomly divided into control group and RSV infected group. Under 10% chloral hydrate 3 渭 l / g anesthesia, the airway hyperresponsiveness was evaluated by 1.5 脳 108PFU/ml RSV or 100 渭 l DMEM medium on the 3rd day and 14th day after nasal drip. The total number of cells in bronchoalveolar lavage fluid (BALF) and white blood cell count (WBC) in bronchoalveolar lavage fluid (BALF) were detected by HE staining. The level of cytokines in alveolar lavage fluid was detected by ELSIA method. The mRNA expression of cholinergic receptor M2 (M 2R) and Toll like receptor 3 (TLR3) in nude mice was detected by Q-PCR on day 5. Results the replication duration was longer and the virus titer was higher in nude mice after infection. The number of cells in bronchoalveolar lavage fluid (BALF) and the pathological score of pulmonary inflammation in nude mice infected with RSV and BALB/c RSV were significantly higher than those in control group (P0.05). The proportion of lymphocytes in bronchoalveolar lavage fluid increased, and the proportion of macrophages decreased. He staining showed that the inflammation of lung was mainly lymphocytic infiltration, thickening of blood vessel wall, bronchial wall and alveolar septum. Inflammation after BALB/cRSV infection lasted longer than RSV infection in nude mice. Lung function test of nude mice and BALB/c mice showed that the airway reactivity was higher in BALB/c mice than in the control group, and the response of the 5th weather tract was the highest in BALB/c mice after RSV infection, which was significantly different from that in RSV infected nude mice at the same time (P0.05). The cytokine IFN- 纬 increased significantly after BALB/c RSV infection, but had no significant change after RSV infection in nude mice compared with the control group. The TLR3mRNA of nude mice after RSV infection was significantly higher than that of the control group (P0.05), but IL-4,IL-10 and cholinergic receptor M2 had no significant difference compared with the control group (P0.05). Conclusion: the duration of viral replication in nude mice infected with RSV is longer, but the duration of pulmonary inflammation is shorter than that of BALB/c mice, and the airway reactivity is significantly lower than that of BALB/c mice infected with BALB/c RSV. These results suggest that airway inflammation and airway hyperresponsiveness are not completely parallel to T cells and their cytokines (such as [FN- 纬]) after RSV infection may play an important role in the pathological changes of airway after RSV infection.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.1
本文编号：2243794
[Abstract]:Aim: to establish a nude mouse model of respiratory syncytial virus (RSV) (RSV) infection and to investigate the effect of T cell deficiency on airway inflammation and airway hyperresponsiveness induced by RSV infection. Methods female nude mice and BALB/c mice of 6-8 weeks were randomly divided into control group and RSV infected group. Under 10% chloral hydrate 3 渭 l / g anesthesia, the airway hyperresponsiveness was evaluated by 1.5 脳 108PFU/ml RSV or 100 渭 l DMEM medium on the 3rd day and 14th day after nasal drip. The total number of cells in bronchoalveolar lavage fluid (BALF) and white blood cell count (WBC) in bronchoalveolar lavage fluid (BALF) were detected by HE staining. The level of cytokines in alveolar lavage fluid was detected by ELSIA method. The mRNA expression of cholinergic receptor M2 (M 2R) and Toll like receptor 3 (TLR3) in nude mice was detected by Q-PCR on day 5. Results the replication duration was longer and the virus titer was higher in nude mice after infection. The number of cells in bronchoalveolar lavage fluid (BALF) and the pathological score of pulmonary inflammation in nude mice infected with RSV and BALB/c RSV were significantly higher than those in control group (P0.05). The proportion of lymphocytes in bronchoalveolar lavage fluid increased, and the proportion of macrophages decreased. He staining showed that the inflammation of lung was mainly lymphocytic infiltration, thickening of blood vessel wall, bronchial wall and alveolar septum. Inflammation after BALB/cRSV infection lasted longer than RSV infection in nude mice. Lung function test of nude mice and BALB/c mice showed that the airway reactivity was higher in BALB/c mice than in the control group, and the response of the 5th weather tract was the highest in BALB/c mice after RSV infection, which was significantly different from that in RSV infected nude mice at the same time (P0.05). The cytokine IFN- 纬 increased significantly after BALB/c RSV infection, but had no significant change after RSV infection in nude mice compared with the control group. The TLR3mRNA of nude mice after RSV infection was significantly higher than that of the control group (P0.05), but IL-4,IL-10 and cholinergic receptor M2 had no significant difference compared with the control group (P0.05). Conclusion: the duration of viral replication in nude mice infected with RSV is longer, but the duration of pulmonary inflammation is shorter than that of BALB/c mice, and the airway reactivity is significantly lower than that of BALB/c mice infected with BALB/c RSV. These results suggest that airway inflammation and airway hyperresponsiveness are not completely parallel to T cells and their cytokines (such as [FN- 纬]) after RSV infection may play an important role in the pathological changes of airway after RSV infection.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.1
【参考文献】
相关期刊论文 前1条
1 周娟;王莉佳;崔玉霞;刘雪梅;杨锡强;;裸鼠呼吸道合胞病毒感染的动物模型[J];Virologica Sinica;2006年03期
，本文编号：2243794
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