肠道病毒71型感染人神经母细胞瘤细胞(SH-SY5Y)的miRNA表达谱

目的 研究肠道病毒71型(EV71)感染神经细胞的miRNA表达谱,探讨miRNA在病毒感染神经细胞中的可能作用.方法 建立EV71感染人神经母细胞瘤细胞(SH-SY5Y)模型,收集感染后48 h细胞.以Taqman低密度芯片检测miRNA表达谱,使用实时RT-PCR对芯片结果进行验证并在TargetScan和miRanda网站预测靶基因,采用GO和KEGG分析靶基因功能.结果 成功建立EV71感染SH-SY5Y细胞模型,通过低密度芯片筛选出215种显著升高的miRNA和25种显著下调的miRNA.经过RT-PCR验证,3种miRNA(MiR-10a*、miR-15b*和miR-195)显著下调,7种miRNA(miR-10a、miR-342-5p、miR-483-5p、Let-7b、miR-99a、miR-140-5p和miR-21)显著上调,与芯片结果相符.GO分析显示发展进程和信号调节条目最富集靶基因.KEGG路径分析显示靶基因在肿瘤路径、蛋白水解、Wnt信号传导、黑素形成、粘附连接、MAPK信号通道最富集.结论 EV71感染神经细胞48 h后miRNA表达谱发生改变,10种变化的miRNA靶基因预测在发展进程、信号传导及凋亡中起着重要的作用,可为后期机制研究提供参考.     

百草枯致小鼠肺泡上皮细胞损伤过程中microRNA表达谱分析

目的 研究百草枯(PQ)致小鼠肺泡上皮细胞损伤过程中microRNA(miRNA)表达谱的变化,并预测差异表达的miRNA靶基因和相关的生物学功能.方法 体外培养小鼠肺泡上皮细胞系MLE12细胞,加入不同浓度的PQ,通过CCK-8方法检测细胞活力以及流式细胞术检测细胞凋亡率,最终选择100μ MPQ作为合适的实验浓度.进一步设立对照组和实验组,对照组加入等量PBS的培养液,实验组加入100μ MPQ的培养液,24h后用基因芯片检测两组microRNA表达谱的改变,分析差异表达的miRNA并用生物信息学方法预测靶基因及相关信号调节通路.结果 100μmol/LPQ处理MLE-12细胞24h后,细胞活性下降约50％,细胞凋亡率约15％.基因芯片共筛选出11个差异表达的miRNA,其中1个被上调,10个被下调.预测这些上下调的miRNA靶基因与细胞的线粒体凋亡途径以及DNA的甲基化相关,参与PI3K-Akt、mTOR、RAS、TNF和MAPK等与氧化应激以及凋亡相关的信号通路的调节.结论 PQ对小鼠肺泡上皮细胞的损伤进程中microRNA表达谱发生了改变,这些差异表达的miRNA可能是细胞氧化应激及凋亡的重要调节因素.     

人乳腺癌MCF-7细胞在不同HER-2水平miRNAs表达谱检测

目的：通过对比观察人乳腺癌细胞MCF-7和MCF-7/HER-2中微小RNA( miRNAs)的差异性表达,初步筛选出与HER-2表达相关的miRNAs表达谱。方法培养人乳腺癌细胞MCF-7和MCF-7/HER-2,利用miRNA芯片技术检测两株细胞中miR-NAs的表达。结果利用miRNA微阵列,筛选获得217种与HER-2相关的miRNAs,较正常MCF-7细胞上调的miRNAs有123个,下调的有94个。在差异有显著性的miRNAs中,hsa-miR-141和hsa-miR-1299的靶基因有多种生物学功能。结论获得人乳腺癌MCF-7细胞在不同HER-2水平miRNAs的差异表达谱,为进一步分析HER-2在乳腺癌中的作用奠定基础。     

基于microRNA差异表达探讨恶性纤维组织细胞瘤致病机制

目的 应用芯片表达分析技术探讨miRNA在恶性纤维组织细胞瘤发病机制中的作用.方法 利用miRCURYTM LNA miRNA表达谱芯片技术高通量筛选NMFH1与MSC、HSF、MRC-5和HT-1080细胞系差异表达miRNA,筛选靶基因,分析调控作用机制.结果 筛查NMFH1与4种对照组细胞系差异表达miRNA,选取与所有正常样本对比均下调miR-222、miR-186及miR-933,均上调miR-886-3p、miR-886-5p、miR-183、miR-340及miR-BART7进行qRT-PCR实验,验证芯片数据可靠性.基于生物信息学方法挖掘miRNA靶基因,构建基因调控网络,基于功能组注释信息分析NMFH1风险疾病基因及通路,揭示MFH恶性病变机制.结论 miRNA在肿瘤样本与正常细胞系间显著差异表达,是参与MFH发生发展过程的重要调控因子,发挥负调控基因表达作用,使肿瘤细胞逃避正常生长调控机制,无限增殖和转移,出现恶性表型.     

T淋巴细胞活化相关的microRNA的筛选、靶分子的预测、构建和鉴定

目的: 筛选T细胞活化相关的microRNA并预测其靶分子,通过双荧光素酶实验鉴定这些microRNA的靶分子方法: 利用Exiqon miRNA基因表达谱芯片,以活化的Jurkat细胞(用CD3抗体和CD28抗体双信号活化)为实验组,未活化Jurkat细胞为对照组,分析了二者miRNA表达谱的差异,找出表达水平显著改变的miRNA,并通过实时定量PCR的方法对芯片结果进行验证;构建miRNA的真核表达载体;利用生物信息学方法预测miRNA的靶分子,构建含靶分子3′UTR的荧光素酶报告基因质粒;分别将miRNA基因真核表达质粒与含有靶分子的荧光素酶报告基因质粒共转染HEK293细胞,进行荧光素酶实验来鉴定miRNA的靶分子结果: Exiqon miRNA基因表达谱芯片显示: Jurkat细胞活化后,miR-202*、 miR-33b、 miR-568和miR-576的表达明显下调;实时定量PCR验证结果与芯片结果一致;酶切鉴定与序列测定证实miR-568的真核表达载体构建成功;利用miRanda软件预测miR-568的靶分子,筛选出与T细胞活化密切相关的分子;双荧光素酶实验显示miR-568对NFAT5有比较明显的抑制作用结论: T细胞活化后,miR-202*、 miR-33b、 miR-568和miR-576表达明显下调;NFAT5可能是miR-568作用的一个靶分子     

人疱疹病毒6型感染HSB-2细胞的lncRNA表达谱变化分析

目的运用高通量测序技术检测人疱疹病毒6型(human herpesvirus 6, HHV-6)感染人淋巴细胞系HSB-2后长链非编码RNA(long non-coding RNA, lncRNA)表达谱的改变, 探究lncRNA在HHV-6感染复制中的作用。方法 HHV-6感染HSB-2细胞72 h后, 提取对照细胞和病毒感染细胞的RNA进行测序, 筛选差异表达的lncRNA;通过生物信息学方法对预测的lncRNA靶基因进行GO注释和KEGG信号通路分析, 构建共表达网络图。qRT-PCR检测差异表达lncRNA的变化倍数。结果共筛选到612个显著差异表达lncRNA, 其中420个为表达上调, 192个表达下调。通过对lncRNA靶基因进行GO和KEGG富集分析显示, 差异表达lncRNA的靶基因与表观染色体调控、免疫应答及细胞代谢等生物学过程密切相关。qRT-PCR确定10条上调IncRNAs表达变化趋势与高通量测序数据一致。结论本研究对HHV-6感染HSB-2细胞的lncRNA表达谱进行分析, 为深入探索lncRNA在HHV-6复制增殖及相关疾病中的作用奠定基础。     

高致病性禽流感病毒H5N1安徽株A/Anhui/1/2005感染人肺癌上皮细胞(A549)的差异表达分析

目的 筛选人细胞中与高致病性禽流感病毒致病相关的基因,探讨高致病性禽流感病毒的致病机理.方法 分别用高致病性禽流感病毒安徽株和普通人流感病毒H1N1株分别感染人肺癌上皮细胞,通过人类全基因表达谱芯片技术对不同时段感染细胞进行差异表达分析,筛选出与高致病性禽流感病毒感染相关的候选基因,以实时定量荧光PCR法验证差异表达基因.结果 获得了不同致病性流感病毒感染细胞后的差异表达谱,验证了细胞凋亡通路、mTOR通路中以及与免疫相关的16个基因的差异表达.结论 H5N1感染后与普通人流感病毒H1N1相比具有促进细胞凋亡的趋势.     

乙型肝炎病毒E抗原肝细胞作用蛋白AK026018表达谱芯片的研究

目的 应用基因芯片技术,筛选能被乙型肝炎病毒E抗原（HBeAg）肝细胞作用蛋白AK026018反式调节的靶基因,初步研究该蛋白的生物学功能.方法 应用反转录聚合酶链反应（RT-PCR）技术,从HepG2细胞中扩增编码AK026018蛋白的全基因,构建真核表达载体,转染肝母细胞瘤系HepG2,提取总mRNA,逆转录为cDNA,与转染空白表达载体pcDNA3.1的HepG2细胞进行DNA芯片分析并比较.结果 经限制性内切酶分析和DNA序列测定鉴定构建的重组表达载体正确.在8464个基因表达谱的筛选中,发现有122个基因有差异表达,其中78种基因表达水平显著下调,45种基因表达水平显著上调.结论 成功地应用DNA芯片技术筛选出HBeAg结合蛋白新基因AK026018的反式调节蛋白,证明该基因对于肝细胞基因表达谱有显著影响.     

永久性房颤的转录组分析及miRNA调控网络的建立

目的探索永久性房颤(p AF)发病相关的关键miRNAs及其调控的靶基因。方法联合应用表达谱芯片和miRNA芯片分析p AF患者(n=7)和健康成人(n=4)的左房组织,筛选p AF相关差异表达的miRNAs,进行靶基因预测后,与表达谱芯片的筛选结果进行负相关分析后的基因集合进行显著性功能分析(GO-analysis);利用miRNA与靶基因之间的靶向调控关系,构建差异miRNA与交集靶基因的调控网络(miRNA-gene-network),得到网络中起核心调控作用的miRNA和被调控的关键靶基因;采用RT-q PCR方法检验另一组p AF患者(n=5)和健康成人(n=4)的左房组织标本。结果表达谱基因芯片发现610个mRNA有显著性改变(fold change2,P0.05),miRNA-靶基因调节网络发现与p AF显著相关的20个miRNAs和107个靶基因,相关度最高的是miR-144、miR-1284、miR-1827、miR-1、miR-3613-3p和miR-101;其调控的重要靶基因包括CACNB2、EFNB1、PTEN、TAOK1、RUNX1和TPM3等;RT-q PCR验证结果显示这些miRNAs和靶基因密切相关。结论通过表达谱基因芯片与miRNAs芯片联合分析p AF左房组织标本,构建miRNA调控网络,发现p AF重要的miRNA-靶基因调控及功能,结果更加准确。     

人CD4~+T淋巴细胞活化相关miRNA的筛选及其靶分子鉴定

目的:筛选与CD4~+ T淋巴细胞活化相关的miRNA,确定其靶分子.方法:用抗CD3的单免隆抗体(mAb)和抗CD28的mAb诱导Jurkat细胞活化,利用miRNA芯片检测并筛选出活化前后表达丰度有显著变化的microRNA,用定量RT-PCR证实筛选出的miRNA在Jurkat细胞活化的差异表达.设计引物构建miRNA的表达载体.利用牛物信息学软件预测miRNA的靶分子,将靶分子mRNA的3'UTR克隆入pGL3Luciferase报告基因载体中.观察转染的miRNA对报告基因表达的影响.结果:流式细胞术(FCM)证实了抗CD3的mAb和抗CD28的mAb可以诱导Jurkat细胞的活化.经miRNA芯片检测表明,Jurkat细胞活化前后有5种miRNA表达水平明显下降,定量RT-PCR证实了miRNA芯片的结果.成功构建了部分预测靶分子的荧光素酶报告基因载体.双荧光素酶报告系统检测了miR-181c对靶分子的干扰作用.结论:miR181c与人CD4~+ T淋巴细胞的活化有着密切的关系,CD69可能足miR-181c的一个靶分子.     

Shrimp miRNAs regulate innate immune response against white spot syndrome virus infection

MicroRNAs are short noncoding RNAs of RNA interference pathways that regulate gene expression through partial complementary base-pairing to target mRNAs. In this study, miRNAs that are expressed in white spot syndrome virus (WSSV)-infected Penaeus monodon, were identified using next generation sequencing. Forty-six miRNA homologs were identified from WSSV-infected shrimp hemocyte. Stem-loop real-time RT-PCR analysis showed that 11 out of 16 selected miRNAs were differentially expressed upon WSSV infection. Of those, pmo-miR-315 and pmo-miR-750 were highly responsive miRNAs. miRNA target prediction revealed that the miRNAs were targeted at 5′UTR, ORF, and 3′UTR of several immune-related genes such as genes encoding antimicrobial peptides, signaling transduction proteins, heat shock proteins, oxidative stress proteins, proteinases or proteinase inhibitors, proteins in blood clotting system, apoptosis-related proteins, proteins in prophenoloxidase system, pattern recognition proteins and other immune molecules. The highly conserved miRNA homolog, pmo-bantam, was characterized for its function in shrimp. The pmo-bantam was predicted to target the 3′UTR of Kunitz-type serine protease inhibitor (KuSPI). Binding of pmo-bantam to the target sequence of KuSPI gene was analyzed by luciferase reporter assay. Correlation of pmo-bantam and KuSPI expression was observed in lymphoid organ of WSSV-infected shrimp. These results implied that miRNAs might play roles as immune gene regulators in shrimp antiviral response.     

人树突状细胞体外经HBsAg刺激后的抗病毒作用

目的观察人树突状细胞(DCs)经表面抗原(HBsAg)刺激、体外诱导自身特异性T淋巴细胞增殖后,对2.2.15细胞中HBeAg和HBsAg的特异性免疫抑制作用。方法 用粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白介素-4(IL-4)和肿瘤坏死因子(TNF-a)分化、诱导人外周血PBMC中的DCs,在DCs成熟前加入纯的HBsAg刺激,将成熟后的DCs体外与自身T淋巴细胞共培养,同时不加HBsAg刺激的DCs与T细胞共培养、T细胞加纯的HBsAg共培养以及单纯T细胞作对照,5 d后收集T细胞,分组加入2.2.15细胞培养液中,分别收集第1天、3天、5天和7天的培养上清液,检测其HBeAg和HBsAg的分泌情况。结果经抗原刺激后的DCs可以有效提呈病毒抗原,正常人与慢性乙肝患者负载抗原后的DCs刺激T淋巴细胞增殖的能力[cpm分别为(46 700±7 850)和(38 628±5 427)]明显高于未负载抗原的DCs[cpm分别为(40 450±4 645和33 924±4 498)]及对照组PBMC[cpm分别为(5 947±476)和(5 089±233)],P<0.01。负载抗原的DCs有强烈的免疫应答活性,并且其免疫刺激能力似乎与负载的抗原量成正比;经抗原刺激激活的T细胞可以有效地抑制HBeAg的表达,但对HBsAg未发现有明显的抑制作用。结论体外经HBsAg刺激后的DCs可有效地提呈病毒抗原,并可进一步激活T细胞产生,同时能显著地抑制2.2.15细胞上清     

Plasmacytoid,conventional, and monocyte‐derived dendritic cells undergo a profound and convergent genetic reprogramming during their maturation

DCs express receptors sensing microbial, danger or cytokine signals, which when triggered in combination drive DC maturation and functional polarization. Maturation was proposed to result from a discrete number of modifications in conventional DCs (cDCs), in contrast to a cell‐fate conversion in plasmacytoid DCs (pDCs). cDC maturation is generally assessed by measuring cytokine production and membrane expression of MHC class II and co‐stimulation molecules. pDC maturation complexity was demonstrated by functional genomics. Here, pDCs and cDCs were shown to undergo profound and convergent changes in their gene expression programs in vivo during viral infection. This observation was generalized to other stimulation conditions and DC subsets, by public microarray data analyses, PCR confirmation of selected gene expression profiles, and gene regulatory sequence bioinformatics analyses. Thus, maturation is a complex process similarly reshaping all DC subsets, including through the induction of a core set of NF‐κB‐ or IFN‐stimulated genes irrespective of stimuli.     

Identification and characterization of Cynoglossus semilaevis microRNA response to Vibrio anguillarum infection through high-throughput sequencing

MicroRNAs (miRNA) play key regulatory roles in diverse biological processes. Cynoglossus semilaevis is an important commercial mariculture fish species in China. To identify miRNAs and investigate immune-related miRNAs of C. semilaevis, we performed high-throughput sequencing on three small RNA libraries prepared from C. semilaevis immune tissues (liver, head kidney, spleen, and intestine). One library was prepared under normal conditions (control, CG); two were prepared during Vibrio anguillarum infection, where vibriosis symptoms were obvious and non-obvious (HOSG and NOSG, respectively). We obtained 11,216,875, 12,313,404, and 11,398,695 clean reads per library, respectively. Bioinformatic analysis identified 452 miRNAs, including 24 putative novel miRNAs. We analyzed differentially expressed miRNAs between two libraries using pairwise comparison. For NOSG–CG, there was significant differential expression of 175 (38.72%) miRNAs. There was significant differential expression of 215 (47.57%) miRNAs between HOSG and CG. Compared with CG, The HOSG–NOSG comparison revealed significantly different expression of 122 (26.99%) miRNAs respectively. Real-time quantitative PCR (RT-qPCR) experiments were performed for 10 miRNAs of the three samples, and agreement was found between the sequencing and RT-qPCR data. For miRNAs that were significantly differentially expressed, functional annotation of target genes by Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that a set of miRNAs that were expressed highly abundantly and significantly differentially were might involved in immune system development and immune response. To our understanding, this is the first report of comprehensive identification of C. semilaevis miRNAs being differentially regulated in immune tissues (liver, head kidney, spleen, and intestine) in normal conditions relating to V. anguillarum infection. Many miRNAs were differentially regulated upon pathogen exposure. This work provides an opportunity for further understanding of the molecular mechanisms of miRNA regulation in C. semilaevis host–pathogen interactions.     

Differential Expression Profiling and Functional Analysis of microRNAs through Stage I-III Papillary Thyroid Carcinoma

Objective: To elucidate the mechanisms undergoing the pathogenesis of PTC, this study try to find stage specific microRNAs (miRNAs) using microarray chip in stage I, II and III papillary thyroid carcinoma (PTC) tissues as well predict miRNAs binding target genes and their molecular functions.Methods: PTC specimens of stage I, II, and III and their paired adjacent non-tumor tissue (one patient for each stage) were collected. The expressions of miRNAs were examined using miRNA microarray chip. The most significant changed miRNAs from microarray were verified by using quantitative RT-PCR. The Potential miRNAs regulating target genes and their preliminary biological functions were forecasted with variety function prediction software.Results: Ten miRNAs exhibited sequential up regulation expression profiles and five miRNAs performed sequential down regulation throughout stage I to III (p<0.05). After normalization, Fifteen miRNAs showed significant different compared to adjacent non-tumor tissues (p<0.05). Among of them, the most significant up regulation and down regulation miRNAs were miR-146b-5p and miR-335, respectively. Both of them were verified with qRT-PCR. 34 target genes for miR-146-5p and 36 target genes for miR-335 was predicted.Conclusion: MicroRNA profile assay successfully detected a branch of differential expression miRNAs between PTC and normal tissue. Some of them also showed stage specific. Biological function analysis showed that target genes were involved in five aspects including cell proliferation, differentiation, apoptosis, cycle, and signaling transduction pathway, suggesting the regulatory role of abnormal expression of critical miRNAs in the pathogenesis of PTC.     

抗原活化慢性乙型肝炎患者外周血树突状细胞的对比研究

目的 探讨不同抗原在体外活化慢性乙型肝炎患者外周血树突状细胞(DC)及诱导特异性T细胞应答的能力.方法 用无血清培养基从慢性乙型肝炎患者外周血中分离培养DC,在DC成熟前,分别加入HBsAg多肽、HBcAg多肽刺激,用流式细胞仪检测DC表型,用液闪计数仪观察DC对T细胞的增殖作用,用ELISA法检测混合淋巴细胞反应(MLR)中IL-12的分泌水平.结果 经HBcAg多肽刺激DC的CD86表达率为(92.20±5.18)%,明显高于HBsAg多肽刺激组(76.19±3.90)%和未加抗原组(62.37±4.24)%,P＜0.01;经HBcAg多肽刺激组DC诱导同种异体静止T细胞增殖的能力每分钟液闪计数值cpm为34 326±3088,明显高于HBsAg多肽刺激组20 306±2897和单个核细胞组3454±409,P＜0.01;经HBcAg多肽刺激组DC MLR中IL-12(348±42.8)ng/L,分别高于HBsAg多肽刺激组(226±30.6)ng/L和未加抗原组(116±15.6)ng/L,P＜0.01.结论 使用HBcAg多肽刺激DC可比HBsAg多肽更有效地提呈病毒抗原,提高诱导特异性T细胞应答的能力.     

Regulation of immune responses and tolerance: the microRNA perspective

Much has been learned about the molecular and cellular components critical for the control of immune responses and tolerance. It remains a challenge, however, to control the immune response and tolerance at the system level without causing significant toxicity to normal tissues. Recent studies suggest that microRNA (miRNA) genes, an abundant class of non-coding RNA genes that produce characteristic approximately 22 nucleotides small RNAs, play important roles in immune cells. In this article, we discuss emerging knowledge regarding the functions of miRNA genes in the immune system. We delve into the roles of miRNAs in regulating signaling strength and threshold, homeostasis, and the dynamics of the immune response and tolerance during normal and pathogenic immunological conditions. We also present observations based on analyzes of miR-181 family genes that indicate the potential functions of primary and/or precursor miRNAs in target recognition and explore the impact of these findings on target identification. Finally, we illustrate that despite the subtle effects of miRNAs on gene expression, miRNAs have the potential to influence the outcomes of normal and pathogenic immune responses by controlling the quantitative and dynamic aspects of immune responses. Tuning miRNA functions in immune cells, through gain- and loss-of-function approaches in mice, may reveal novel approach to restore immune equilibrium from pathogenic conditions, such as autoimmune disease and leukemia, without significant toxicity.     

蜂胶黄酮PB3A作用下结肠癌细胞株中miRNA-198和miRNA-296-5p的表达及功能预测

目的:探讨蜂胶黄酮短叶松素-3-乙酸酯(pinobanksin-3-acetate,PB3A)对结肠癌SW480细胞微小RNA(microRNA,miRNA)表达谱的影响,为结肠癌的治疗及靶向药物研发提供理论依据。方法:使用miRNA芯片技术分析检测蜂胶黄酮PB3A处理人类结肠癌SW480细胞后miRNA表达谱的变化。通过实时荧光定量PCR方法检测miRNA-198和miRNA-296-5p的表达水平,以此来验证miRNA芯片结果的准确性和可靠性。利用miRWalk、Micro T、miRanda等12个网上数据库预测这2条miRNAs的靶基因并进行靶基因功能富集分析。结果:miRNA芯片分析结果显示,蜂胶黄酮PB3A干预24 h后结肠癌SW480细胞中差异表达倍数在2倍及以上的miRNA有267条,其中差异表达倍数达10倍及以上的miRNA有30条,28条为上调表达,2条下调表达;RT-qPCR实验结果显示miRNA-198和miRNA-296-5p的表达量趋势跟miRNA芯片结果一致,表达差异有统计学意义(P0.05)。miRNA靶基因预测发现miRNA-198有859个靶基因,miRNA-296-5p有906个靶基因;对这些可能被调控的靶基因进行Gene Class分析,结果显示miRNA-198和miRNA-296-5p的靶基因功能主要为转录因子、拷贝数变异、细胞分化、癌基因、蛋白激酶、组蛋白、转移癌基因、肿瘤抑制基因等(P0.05)。信号通路富集分析结果显示,miRNA-198靶基因显著富集于肿瘤通路、Wnt信号通路、胞吞通路、Erb B信号通路、黏着斑通路、黑素生成通路等信号通路,而miRNA-296-5p靶基因在MAPK信号通路、胞吞通路、轴突导向通路、Wnt信号通路、胰岛素信号通路、钙离子信号通路等信号通路中出现聚集(P0.05)。结论:蜂胶黄酮PB3A影响结肠癌SW480细胞的miRNA表达谱。PB3A作用下miRNA-198和miRNA-296-5p的异常表达可能参与PB3A抗结肠癌的过程。     

闭塞性细支气管炎模型大鼠微小RNA差异表达谱分析

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。 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：