RSV感染幼龄大鼠肺组织TSLP与Th2反应的相关性分析

Correlative analysis of TSLP and Th2 responses in lung tissue of young rats infected with RSV

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(P<0.05);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(P<0.05);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。

Objective: To investigate the changes of thymic lymphocyte procyanidin( TSLP) and interleukin-4( IL-4),interleukin-10( IL-10) and interleukin-13( IL-13) in the lung tissue of young rats infected with respiratory syncytial virus,and to investigate the changes of TSLP and Th2 Correlation. Methods: 18 Wistar rats were randomly divided into RSV infection group and normal group,9 rats in each group. Application of RSV virus droplets nasal modeling,saline diarrhea as a control. The lung tissue of rats was taken for pathological observation; the load of syncytial virus at different time points and the expression of TSLP in lung tissue were detected by Real-time quantitative polymerase chain reaction( q-PCR); at the same time,the expression of TSLP at the protein level was detected by Western blot. The levels of cytokines IL-4,IL-10 and IL-13 in serum were measured by enzyme-linked immunosorbent assay( ELISA). And the relationship between TSLP and IL-4,IL-10 and IL-13 was analyzed by Spearman correlation. Results: Compared with the normal control group,the rats in the model group showed symptoms such as poor mental state,rough hair,decreased activity and shortness of breath after intranasal infection,and the symptoms were aggravated. Pathological observation showed that the alveolar wall was thickened in the model group,and a large number of lymphocytes,plasma cells,eosinophil infiltration. q-PCR was used to detect the syncytial virus load in model group,and reached the peak at about 5 days,then decreased gradually. RSV infection increased the secretion of TSLP in rat respiratory tract. q-PCR and Western blot showed that TSLP concentration in model group was higher than that in control group( P<0. 05). The levels of IL-4,IL-10 and IL-13 in the serum of the model group were higher than those in the control group( P<0. 05). Spearman correlation analysis showed that TSLP was positively correlated with IL-4 and IL-10 expression in RSV model group,and there was no correlation between TSLP and IL-13 expression. Conclusion: RSV infection in rat lung tissue can induce increased expression of TSLP,while promoting the occurrence of Th2 type of inflammatory response and the expression of related cytokines,in order to further study of RSV infection caused by bronchiolitis and asthma and other respiratory diseases,the mechanism of action and clinical treatment laid the foundation.