摘要
目的:研究反复呼吸道合胞病毒(RSV)感染大鼠脊髓背根节感觉神经细胞转录因子的活化状态变化。方法:1～2周龄SD大鼠幼鼠共80只,区组随机分为对照组和RSV感染组,每组40只。RSV感染组采用每周1次RSV(浓度5×105U/mL)滴鼻法制作RSV慢性感染模型,对照组采用无病毒培养上清液滴鼻。8周后每组取5只大鼠进行气道反应性测定。原位杂交检测肺组织RSVRNA以证明呼吸道合胞病毒感染模型的可靠性。用TranSignal蛋白质/DNA活性芯片筛选C7～T5脊髓背根节组织中活性改变的转录因子。Western印迹对筛选结果中改变最明显的因子进行验证。结果:RSV感染组气道阻力增幅显著大于对照组(P<0.05)。原位杂交显示RSV感染组肺组织有大量呼吸道合胞病毒存在,HE染色示肺部存在明显炎症细胞浸润,说明呼吸道合胞病毒感染模型建立成功。TranSignal蛋白/DNA组合芯片检测筛选:RSV反复感染大鼠背根节55个转录因子活性上调(RSV感染组/对照组>2),43个活性下调(RSV感染组/对照组<0.5)。Western印迹验证结果与芯片结果一致,并进一步明确蛋白/DNA组合芯片筛选的IRF转录因子是IRF-1,而不是IRF-2。结论:反复RSV感染可引起气道反应性增高和脊髓背根节感觉神经细胞内多种转录因子活性发生改变,可能与气道反应性增高的神经调控异常有关。
Objective To elucidate intracellular transcription factor activation of C7T5 dorsal root ganglia in rats recurrently infected with respiratory syncytial virus（RSV）.Methods Eighty 12 weeks old Sprague-Dawley rats were randomly divided into 2 groups：a control group and a RSV-infection group.The rats in the RSV-infection group were infected with 5 ×10^5 U/mL RSV once a week and the rats in the control group were treated with culture medium without RSV.Airway response was measured after 8 weeks.Lung tissue was submitted for HE staining and in situ hybridization.The C7T5 dorsal root ganglia were obtained for the preliminary screening of the intracellular transcription factors by TranSignalTM protein/DNA combo array.Nuclear protein of C7T5 dorsal root ganglia were extracted and submitted to Western blot.Results Airway response in the RSV-infection group was higher than that in the control group（P〈0.05）.HE staining showed inflammatory cell infiltration,and in situ hybridization demonstrated positive RSV RNA in the RSV-infection rat lung which was not present in the control group,thus validating the efficacy of our model.TranSignalTM protein/DNA combo array screening showed that 55 transcription factors increased by at least 2 folds in the C7T5 DRG cells of the RSV-infection group.The transcription factors Smad and interferon regulatory factor（1 or 2）were the 2 most upregulated transcription factors identified by combo array screening（59 and 43 fold increase compared with the control,respectively）.Western blot confirmed Smad（1/2/3）and IRF-1 upregulate while IRF-2 remained unchanged.Conclusion Respiratory syncytial virus infection results in airway hyperresponsiveness and transcription factor activation in C7T5 spinal adorsal root ganglia in rats,which may contribute to airway nerve network dysfunction and airway hyperresponsiveness.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2009年第12期1189-1195,共7页
Journal of Central South University ：Medical Science
基金
国家自然科学基金(30670914)~~
