肾移植受者外周血单个核细胞中miR-29与急性排斥反应的相关性研究

Correlation between microRNA-29 family molecules in peripheral blood mononuclear cells and allograft acute rejection after renal transplantation

目的了解肾移植受者外周血单个核细胞（PBMC）中的微小RNA（miRNA-29（miR-29）家族分子在急性排斥反应（AR）发生及恢复后的变化。方法15例肾移植后发生AR的受者，分别在AR发生时（AR组）和恢复后（恢复组）采集外周血，提取PBMC的总RNA，共采集30个样本。用实时荧光定量聚合酶链反应方法分别对miR-29a、miR-29b和miR-29c的表达水平进行相对定量。通过PicTar和TargetScan预测miR-29调控的靶基因，再通过组织特异性数据库，对血液中miRNA靶基因进行筛查过滤。对miR-29对应的靶基因进行基因本体论（G0）和京都基因与基因组百科全书（KEGG）通路分析，得到miR-29可能的生物学过程和作用通路。结果miR-29a和-29c在恢复组的表达水平明显高于AR组（P〈0．05），而两组miR-29b的表达的差异无统计学意义（P〉0．05）。GO中，有14个生物学过程在预测的靶基因中被有意义的富集（P〈0．05）；在KECG中，3条途径被有效富集（P〈0．05）。结论miR-29通过调控代谢、生物学合成等生物学过程和（或）黏着斑通路等影响AR的恢复。受者PBMC中miR-29的检测未来可能会成为判断AR后移植。肾转归的微创检测方法。

Objective To investigate whether acute rejection is associated with significant alterations in the expression of microRNA-29 （miR-29） family members in the peripheral blood mononuclear cells （PBMCs）. Methods A total of 15 independent transplantation recipients were involved in this study. Two time points, AR onset and AR reversal, were adopted for each recipient. The quantitative real-time PCR （qRT-PCR） was conducted for the relative quantification of miR-29a, -29b and -29c. The target genes regulated by miR-29 were analyzed by using bioinformatics. Results miR-29a and -29c were differentially expressed in the AN reversal phase compared to the AN onset in the qRT-PCR assays, while there were no significant difference in the expression of miR-29b between the two groups. A total of 14 GO （gene ontology） terms in biological processes were significantly enriched, and 3 KEGG pathways were overrepresented （P〈0. 05）. Conclusion miR-29a and -29c are functional in the recovery period after AN and may be useful molecular markers for monitoring the recovery period of AR. The probable biochemical processes and pathways of miR-29 associated with acute rejection reversal may be ＇regulation of metabolic process＇, ＇biosynthetic process＇ and/or ＇focal adhesion＇.