闭塞性细支气管炎模型大鼠微小RNA差异表达谱分析

摘要

BACKGROUND:There is no effective therapy for obliterative bronchiolitis after tracheal transplantation. A therapeutic strategy at microRNA (miRNA) molecular level plays a crucial role in the prevention and treatment of complications after organ transplantation. OBJECTIVE:To analyze the miRNA differential expression profile in response to obliterative bronchiolitis after orthotopic tracheal transplantation in rats. METHODS:The obliterative bronchiolitis model after lung transplantation was established through orthotopic tracheal transplantation in inbred strains of rats, and then was identified using histoIogical examination. Total miRNAs were detected by miRNA array and significantly differential expressed miRNAs were filtrated in the transplanted trachea tissues. The miRNA-146a, miRNA-155 and miRNA-451 with significantly differential expressions were used for relative quantitative study. Quantitative real-time reverse transcription-polymerase chain reaction was applied to verify the reliability of miRNA array results. RESULTS AND CONCLUSION:The pathological examination showed that, obliterative bronchiolitis model in rats was successful y established at 4 weeks after orthotopic tracheal transplantation. A total of obliterative bronchiolitis-related 29 miRNAs were found in miRNA expression profiles, including 14 miRNAs with significantly down-regulated expression and 15 miRNAs with significantly up-regulated expression. Among them, the significantly up-regulated miRNAs (miRNA-146a and miRNA-155) and the significantly down-regulated miRNA-451 were involved in immuno-inflammatory reaction. The miRNAs play an important role in regulating pathophysiological changes of obliterative bronchiolitis after lung transplantation.%背景：目前对于气管移植后并发闭塞性细支气管炎尚无有效的治疗方法。miRNA分子水平机制的治疗策略在防治器官移植后并发症方面具有重要意义。　　目的：分析大鼠原位气管移植模拟肺移植后发生闭塞性细支气管炎的微小RNA 表达谱变化。　　方法：通过近交系大鼠原位气管移植，模拟建立肺移植闭塞性细支气管炎的动物模型并经病理学证实；通过微小 RNA芯片筛选出供体移植气管闭塞性细支气管炎组织中显著差异表达的微小 RNA，并选取差异表达显著的miR-146a、miR-155和miR-451进行相对定量研究，应用实时定量RT-PCR(RT-qPCR)法验证芯片结果的可靠性。　　结果与结论：原位气管移植后4周经病理检查证实大鼠闭塞性细支气管炎模型成功建立。microarray检测获得29个与闭塞性细支气管炎相关的微小RNA，包括15个微小RNA表达显著上调，14个微小RNA表达显著下调，其中显著上调miR-146a、miR-155和显著下调miR-451的功能涉及免疫炎症反应等。提示微小RNA在肺移植后闭塞性细支气管炎病理进程中发挥着重要的调控作用。