Ge Qin-Min, Huang Chun-Mei, Zhu Xiang-Yang, Bian Fan, Pan Shu-Ming
Department of Emergency, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
Division of Nephrology and Hypertension, Department of Internal Medicine, Mayo Clinic College of Medicine, Rochester, MN, United States of America.
PLoS One. 2017 Mar 15;12(3):e0173292. doi: 10.1371/journal.pone.0173292. eCollection 2017.
To identify specific miRNAs involved in sepsis-induced AKI and to explore their targeting pathways.
The expression profiles of miRNAs in serum from patients with sepsis-induced AKI (n = 6), sepsis-non AKI (n = 6), and healthy volunteers (n = 3) were investigated by microarray assay and validated by quantitative PCR (qPCR). The targets of the differentially expressed miRNAs were predicted by Target Scan, mirbase and Miranda. Then the significant functions and involvement in signaling pathways of gene ontology (GO) and KEGG pathways were analyzed. Furthermore, eight miRNAs were randomly selected out of the differentially expressed miRNAs for further testing by qPCR.
qPCR analysis confirmed that the expressions levels of hsa-miR-23a-3p, hsa-miR-4456, hsa-miR-142-5p, hsa-miR-22-3p and hsa-miR-191-5p were significantly lower in patients with sepsis compared with the healthy volunteers, while hsa-miR-4270, hsa-miR-4321, hsa-miR-3165 were higher in the sepsis patients. Statistically, miR-4321; miR-4270 were significantly upregulated in the sepsis-induced AKI compared with sepsis-non AKI, while only miR-4321 significantly overexpressed in the sepsis groups compared with control groups. GO analysis showed that biological processes regulated by the predicted target genes included diverse terms. They were related to kidney development, regulation of nitrogen compound metabolic process, regulation of cellular metabolic process, cellular response to oxidative stress, mitochondrial outer membrane permeabilization, etc. Pathway analysis showed that several significant pathways of the predicted target genes related to oxidative stress. miR-4321 was involved in regulating AKT1, mTOR and NOX5 expression while miR-4270 was involved in regulating PPARGC1A, AKT3, NOX5, PIK3C3, WNT1 expression. Function and pathway analysis highlighted the possible involvement of miRNA-deregulated mRNAs in oxidative stress and mitochondrial dysfunction.
This study might help to improve understanding of the relationship between serum miRNAs and sepsis-induced AKI, and laid an important foundation for further identification of the potential mechanisms of sepsis-induced AKI and oxidative stress and mitochondrial dysfunction.
鉴定参与脓毒症诱导的急性肾损伤（AKI）的特定微小RNA（miRNA），并探索其靶向途径。
通过微阵列分析研究脓毒症诱导的AKI患者（n = 6）、非脓毒症AKI患者（n = 6）和健康志愿者（n = 3）血清中miRNA的表达谱，并通过定量聚合酶链反应（qPCR）进行验证。通过Target Scan、mirbase和Miranda预测差异表达miRNA的靶标。然后分析基因本体论（GO）和京都基因与基因组百科全书（KEGG）途径的显著功能及在信号通路中的参与情况。此外，从差异表达的miRNA中随机选择8个miRNA通过qPCR进行进一步检测。
qPCR分析证实，与健康志愿者相比，脓毒症患者中hsa-miR-23a-3p、hsa-miR-4456、hsa-miR-142-5p、hsa-miR-22-3p和hsa-miR-191-5p的表达水平显著降低，而脓毒症患者中hsa-miR-4270、hsa-miR-4321、hsa-miR-3165表达较高。统计学上，与非脓毒症AKI相比，脓毒症诱导的AKI中miR-4321和miR-4270显著上调，而与对照组相比，脓毒症组中只有miR-4321显著过表达。GO分析表明，预测靶基因调控的生物学过程包括多种术语。它们与肾脏发育、氮化合物代谢过程调节、细胞代谢过程调节、细胞对氧化应激的反应、线粒体外膜通透性等有关。通路分析表明，预测靶基因与氧化应激相关的几个重要通路有关。miR-4321参与调节AKT1、mTOR和NOX5的表达，而miR-4270参与调节PPARGC1A、AKT3、NOX5、PIK3C3、WNT1的表达。功能和通路分析突出了miRNA失调的mRNA可能参与氧化应激和线粒体功能障碍。
本研究可能有助于增进对血清miRNA与脓毒症诱导的AKI之间关系的理解，并为进一步鉴定脓毒症诱导的AKI以及氧化应激和线粒体功能障碍的潜在机制奠定重要基础。
