RSV感染幼龄大鼠肺组织TSLP与Th2反应的相关性分析

发布时间：2018-05-26 15:02

本文选题：呼吸道合胞病毒(RSV) + 胸腺基质淋巴细胞生成素(TSLP)　；参考：《中国免疫学杂志》2017年09期

【摘要】：目的:通过观察当呼吸道合胞病毒(RSV)感染幼龄大鼠肺组织时胸腺基质淋巴细胞生成素(TSLP)与IL-4、IL-10和IL-13的变化,探讨TSLP与Th2反应的相关性。方法:将18只正常Wistar大鼠随机分为模型组和正常对照组,每组各9只。应用RSV病毒液滴鼻造模,生理盐水滴鼻做对照。取大鼠肺部组织进行病理切片观察炎症反应情况;运用实时荧光定量聚合酶链式反应(q-PCR)分别检测两组大鼠不同时间点合胞病毒载量情况和肺组织TSLP在基因水平的表达情况,同时采用免疫印记法(Western blot)检测肺组织TSLP在蛋白水平的表达情况;同时应用酶联免疫吸附试验(ELISA)检测大鼠血清中Th2型相关细胞因子IL-4、IL-10和IL-13的表达水平;并对TSLP与L-4、IL-10、IL-13的关系进行Spearman相关分析。结果:与正常对照组相比,模型组大鼠在滴鼻感染后出现精神状态差、毛粗糙、活动减少、呼吸短促等症状,且上述症状进行性加重;病理观察显示模型组大鼠可见肺泡壁明显增厚,肺间质可见大量的浆细胞、淋巴细胞以及嗜酸性粒细胞浸润;q-PCR法检测模型组大鼠合胞病毒载量逐渐增加,在第5天左右达到峰值,后逐渐降低;RSV感染促进大鼠体内TSLP表达水平增加,q-PCR法和Western blot法检测显示:模型组大鼠的TSLP浓度均高于对照组(P0.05);同时ELISA检测表明模型组大鼠血清中IL-4、IL-10和IL-13的浓度均高于对照组(P0.05);Spearman相关分析显示RSV模型组大鼠的TSLP与IL-4和IL-10表达水平均呈正相关,TSLP与IL-13表达水平无相关性。结论:RSV感染Wistar大鼠肺组织时,可诱导TSLP表达水平增加,同时促进Th2型炎症反应的发生及相关细胞因子的表达,为进一步研究RSV感染致毛细支气管炎和哮喘等喘息类疾病的作用机制及临床治疗奠定了基础。
[Abstract]:Objective: To observe the changes of thymic stromal lymphopoietin (TSLP) and IL-4, IL-10 and IL-13 in the lung tissue of young rats infected with respiratory syncytial virus (RSV), and to explore the correlation between TSLP and Th2 reaction. Methods: 18 normal Wistar rats were randomly divided into model group and normal control group, 9 rats in each group. Model, physiological saline drip nose was used as control. Pathological section of rat lung was taken to observe the inflammatory response. Real-time fluorescence quantitative polymerase chain reaction (q-PCR) was used to detect the load of syncytial virus at different time points and the expression of TSLP in the lung tissue in two groups, and the immune imprint method (Western blot) was used. The expression of TSLP in the protein level of lung tissue was detected; at the same time, enzyme linked immunosorbent assay (ELISA) was used to detect the expression level of Th2 related cytokines IL-4, IL-10 and IL-13 in rat serum, and the relationship between TSLP and L-4, IL-10 and IL-13 was carried out Spearman correlation analysis. The symptoms such as poor mental state, rough hair, reduced activity, short breathing and other symptoms were observed, and the above symptoms were aggravated. Pathological observation showed that the alveolar wall in the model group was obviously thickened, the pulmonary interstitium could be seen in a large number of plasma cells, lymphocyte and eosinophil infiltration, and the load of the syncytial virus increased gradually in the q-PCR test model group. The peak value reached fifth days and then decreased gradually. RSV infection promoted the TSLP expression in rats. The q-PCR and Western blot assay showed that the TSLP concentration in the model group was higher than that of the control group (P0.05), and the ELISA detection showed that the concentration of IL-4, IL-10 and IL-13 in the serum of the model group was higher than that of the control group (P0.05). The correlation analysis showed that the TSLP in the RSV model group was positively correlated with the expression level of IL-4 and IL-10, and there was no correlation between the expression level of TSLP and IL-13. Conclusion: RSV can induce the increase of TSLP expression level in the lung tissue of Wistar rats with RSV infection, and promote the occurrence of the Th2 type inflammatory reaction and the expression of related cytokines, which can further study the capillary capillary of RSV infection. The mechanism and clinical treatment of asthmatic diseases such as bronchitis and asthma have laid the foundation.
【作者单位】：大连市儿童医院;
【基金】：国家自然科学基金(No.81473726) 辽宁省自然科学基金项目(2014020173)资助
【分类号】：R725.6

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