Transcriptomic analysis and laboratory experiments reveal potential critical genes and regulatory mechanisms in sepsis-associated acute kidney injury.
Transcriptomic analysis and laboratory experiments reveal potential critical genes and regulatory mechanisms in sepsis-associated acute kidney injury.
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转录组分析和实验室实验揭示脓毒症相关急性肾损伤的潜在关键基因和调节机制
DOI：
10.21037/atm-22-845
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发表时间：
2022-07
影响因子：
--
通讯作者：
Chi, Xinjin
中科院分区：
文献类型：
--
作者：
Liu, Boyang;Ao, Shengxiang;Tan, Fang;Ma, Wei;Liu, Haoru;Liang, Huaping;Yang, Xia;Chi, Xinjin
关键词：
脓毒症相关急性肾损伤（SA-AKI）是脓毒症最常见和最严重的并发症之一。然而，肾脏病理生理机制的转录调控网络尚未被揭示。这项研究使用生物信息学分析和实验室实验确定了SA-AKI的新机制。我们对盲肠结扎穿孔（CLP）后的小鼠肾脏进行转录组学分析，以模拟临床脓毒症。分离来自CLP和对照组的肾脏样品的RNA，并使用批量信使RNA（mRNA）-seq进行分析。鉴定两组之间的差异表达基因（DEG），并进行GO、KEGG和GSEA途径富集分析。分析了DEG和hub基因的蛋白质-蛋白质相互作用（PPI）网络。使用定量实时聚合酶链反应（qPCR）或蛋白质印迹法验证枢纽基因。通过qPCR、Western blotting、丙二醛（MDA）测定和流式细胞仪分析，预测了hub基因的相互作用网络、靶向microRNA（miRNAs）和长链非编码RNA（lncRNAs），并验证了miRNA-hub基因调控轴。进行N6-腺苷甲基化（m6 A）RNA甲基化调节子和hub基因的相关性分析和m6 A修饰分析。使用高通量测序在两组之间总共鉴定了4，754个DEG。DEG富集的途径包括铁凋亡（最高富集评分）、细胞凋亡和PI 3 K-Akt、NF-κ B和IL-17信号传导途径。15个hub基因中有7个（Hmox 1、Spp 1、Socs 3、Mapk 14、Lcn 2、Cxcl 1和Cxcl 12）参与KEGG通路。mmu-miR-7212- 5 p-Hmox 1是铁凋亡的关键RNA调控轴。m6 A RNA甲基化修饰参与SA-AKI。相关性分析表明m6 A RNA甲基化调控因子与重要的中枢基因之间存在密切的相互作用。这项研究的发现为SA-AKI的发生和发展机制提供了新的见解。mmu-miR-7212- 5 p-Hmox 1轴在铁凋亡和m6 A RNA甲基化调节中的作用可能对未来SA-AKI的治疗具有潜在的临床意义。本研究生成的数据集可在GEO数据库（序列号：GSE 186822）的存储库中找到。
Sepsis-associated acute kidney injury (SA-AKI) is one of the most frequent and serious complications of sepsis. However, the transcriptional regulatory network of the pathophysiological mechanism of the kidney has not been revealed. This study identified new mechanisms in SA-AKI using bioinformatics analyses and laboratory-based experiments. We performed transcriptomic profiling of mouse kidneys after cecal ligation and puncture (CLP) to mimic clinical sepsis. RNA from kidney samples from the CLP and control groups was isolated and analyzed using bulk messenger RNA (mRNA)-seq. Differentially expressed genes (DEGs) between the two groups were identified, and GO, KEGG and GSEA pathway enrichment analyses were performed. The protein-protein interaction (PPI) network of DEGs and hub genes was analyzed. The hub genes were verified using quantitative real-time polymerase chain reaction (qPCR) or Western blotting. The interaction network, targeted microRNAs (miRNAs) and long noncoding RNAs (lncRNAs) of hub genes were predicted, and the critical miRNA-hub gene regulatory axis was verified using qPCR, Western blotting, malondialdehyde (MDA) determination and flow cytometry. Correlation analyses of N6-adenosine methylation (m6A) RNA methylation regulators and hub genes and m6A modification analysis were performed. A total of 4,754 DEGs were identified between the two groups using high-throughput sequencing. The pathways in which DEGs were enriched included ferroptosis (the highest enrichment score), apoptosis, and the PI3K-Akt, NF-kappa B and IL-17 signaling pathways. Seven (Hmox1, Spp1, Socs3, Mapk14, Lcn2, Cxcl1 and Cxcl12) of the 15 hub genes were involved in the KEGG pathway. mmu-miR-7212-5p-Hmox1 was a key RNA regulatory axis in ferroptosis. m6A RNA methylation modifications were involved in SA-AKI. The correlation analyses showed the close interactions among the m6A RNA methylation regulators and important hub genes. The findings of this study provide new insights into the mechanism regulating the occurrence and progression of SA-AKI. The mmu-miR-7212-5p-Hmox1 axis in ferroptosis and m6A RNA methylation regulators may have potential clinical significance for the future treatment of SA-AKI. The datasets generated for this study can be found in the repository of the GEO database (Series number: GSE186822).
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影响因子：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
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作者：
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影响因子：
5.3
作者：
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通讯作者：
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影响因子：
81.5
作者：
Kellum, John A.;Romagnani, Paola;Anders, Hans-Joachim
通讯作者：
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