Differential expression profiles of microRNAs in a rat model of obliterative bronchiolitis

doi:10.3969/j.issn.2095-4344.2014.18.012

Abstract

Abstract:

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 successfully 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.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: trachea, transplantation, lung transplantation, bronchiolitis obliterans

CLC Number:

R318

Wang Jin-yi, Cao Hao, Hong Xuan, Chen Guo-han, Fan Hui-min, Li Qin-chuan, Liu Zhong-min. Differential expression profiles of microRNAs in a rat model of obliterative bronchiolitis[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(18): 2855-2860.

Figures/Tables 1

2.1 实验动物数量分析实验选用供体BN大鼠6只，供体和受体Lewis大鼠18只，分为2组，两组受体大鼠均存活，全部进入结果分析。 2.2 大鼠气管移植一般状况标本采集前各组大鼠均存活。移植后大鼠精神状态稍差，但进食无异常；活动后有轻中度喘鸣，未见合并呼吸衰竭、窒息等征象，亦无感染迹象。 2.3 移植气管组织病理学检查结果实验使用具有典型中期(移植后4周)闭塞性细支气管炎形成的移植气管与同期相应的对照组移植物进行比较。两组气管吻合口愈合良好，其周围为颈部肌肉所覆被，软骨无软化，形态完整。移植气管病理苏木精-伊红染色观察(图1)：对照组移植气管的管壁可见柱状纤毛上皮细胞，内皮轻度增厚，但组织形态上无明显异常或仅为轻度的炎性细胞浸润；闭塞性细支气管炎组移植气管壁的上皮发生异常和/或黏膜腺体消失；气管内膜部分见纤毛柱状上皮细胞生长，黏膜下外周炎性细胞浸润和基质沉积、纤维组织增生，管腔内可有肉芽组织形成，管腔不同程度发生闭塞性细支气管炎，发生率为100%。 2.4 miRNA芯片检测结果通过使用miRNA芯片比较分析移植气管闭塞性细支气管炎组与移植气管对照组的miRNA表达谱，检测miRNA总数为699个。共检测出62个差异表达miRNA，其中33个miRNA移植气管闭塞性细支气管炎组/移植气管对照组表达比值在0.5-2.0之间，差异无显著性意义，占总差异表达miRNA的53.23%。有29个miRNA差异表达显著，占总差异表达miRNA的46.77%，其中14个表达显著下调，占差异表达显著miRNA的48.28%；15个表达显著上调，占差异表达显著miRNA的51.72%(表1)。 2.5 RT-qPCR验证miRNA芯片结果根据miRNA微阵列芯片结果挑选表达显著上调者(miR-146a、miR-155)和表达显著下调者(miR-451)进行相对定量研究，证实与芯片一致。结果表明移植气管闭塞性细支气管炎组miR-146a和miR-155基因表达量分别是移植气管对照组的(3.581± 0.886)倍和(3.163±0.529)倍，提示表达量明显上调，两组差异具有显著性意义(P < 0.05)；移植气管闭塞性细支气管炎组miR-451基因表达量是移植气管对照组的(0.032± 0.012)倍，表达量明显下调，差异有显著性意义(P < 0.05)。利用预测miRNA靶基因常用的TargetScan(http:// targetscan.org/index.html/)数据库预测两组差异表达显著的miRNA靶基因。作为本研究闭塞性细支气管炎组miRNA靶基因的代表，其中miR-146a靶基因是IRAK1和TRAF6，前者与白细胞介素1引起的核因子κB上调有关，后者参与炎症细胞反应；miR-155靶基因为TSHZ3，其参与转录调控；而miR-451的靶基因则是Tollip，与炎症调节有关。"

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