MicroRNA-188 aggravates contrast-induced apoptosis by targeting SRSF7 in novel isotonic contrast-induced acute kidney injury rat models and renal tubular epithelial cells
MicroRNA-188 aggravates contrast-induced apoptosis by targeting SRSF7 in novel isotonic contrast-induced acute kidney injury rat models and renal tubular epithelial cells
复制标题
在新型等渗造影剂诱导的急性肾损伤大鼠模型和肾小管上皮细胞中，MicroRNA-188 通过靶向 SRSF7 加剧造影剂诱导的细胞凋亡
DOI：
10.21037/atm.2019.07.20
复制
发表时间：
2019-08-01
影响因子：
--
通讯作者：
Chen, Jiyan
中科院分区：
文献类型：
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作者：
Liu, Bowen;Chai, Yunfei;Chen, Jiyan
背景资料：造影剂（CM）广泛用于心导管插入术;然而，它可能导致造影剂诱导的急性肾损伤（CI-AKI），严重增加死亡率。microRNA（miRNA）参与了急性肾损伤（阿基）的发生发展过程，这一发现对AKI的诊断和治疗具有巨大的潜力。然而，miRNA在CI-AKI中的作用仍不清楚。本研究旨在探讨miRNAs在CI-AKI中的调控作用。方法：采用临床常用的CM碘克沙醇建立一种新型的、具有代表性的等渗CI-AKI模型。进行下一代测序和逆转录聚合酶链反应（RT-qPCR）以确定CI-AKI中miRNA-188的表达。Western blot分析凋亡调节蛋白和TUNEL法检测细胞凋亡。结果：建立的等渗CI-AKI大鼠模型在血清肌酐、胱抑素C、HE染色及电镜观察等方面具有典型的CI-AKI特征。测序和RT-qPCR结果显示，miRNA-188在CI-AKI大鼠和HK-2细胞模型中均显著上调，而miRNA-188过表达则显著加重CI-AKI细胞模型中的凋亡。SRSF 7被鉴定为miRNA-188的直接靶基因，双荧光素酶报告基因分析确定了SRSF 7与miRNA-188之间的直接相互作用。此外，SRSF 7沉默降低了CI-AKI细胞模型的细胞存活率。结论：本研究的结果表明，miRNA-188通过调节SRSF 7加重了对比剂诱导的细胞凋亡，这可能是CI-AKI干预的潜在药物靶点。
Background: Contrast media (CM) is widely used in cardiac catheterization; however, it may cause contrast-induced acute kidney injury (CI-AKI) which severely increases mortality. MicroRNA (miRNA) has been found to participate in the process of acute kidney injury (AKI), and this discovery has great potential for diagnosis and treatment. However, the role of miRNA in CI-AKI is still unclear. This study aimed to investigate the regulatory effect miRNAs exert in CI-AKI.Methods: We established a novel, representative, isotonic CI-AKI model by using CM iodixanol, a CM which is commonly used in clinic. Next-generation sequencing and reverse transcription polymerase chain reaction (RT-qPCR) were performed to determine the expression of miRNA-188 in CI-AKI. Western blot analysis of the apoptosis regulator protein and TUNEL assay were ordered to evaluate apoptosis. Bioinformatics and double luciferin reporter gene assay were performed to predict and to confirm the interaction between microRNA-188 and SRSF7.Results: The novel isotonic CI-AKI rat model we established exhibited typical characteristics of CI-AKI in serum creatinine, cystatin C, HE staining, and under electron microscope observation. Sequencing and RT-qPCR demonstrated that miRNA-188 was significantly up-regulated both in CI-AKI rat and HK-2 cell models while overexpression of miRNA-188 significantly aggravated apoptosis in CI-AKI cell models. SRSF7 was identified as a direct target gene of miRNA-188, and dual luciferase reporter assay determined the direct interaction between SRSF7 and miRNA-188. In addition, SRSF7 silencing reduced the cell viability rate of the CI-AKI cell model.Conclusions: The present study's findings indicate that miRNA-188 aggravated contrast-induced apoptosis by regulating SRSF7, which may serve as a potential drug target for CI-AKI intervention.