Department of Cardiovascular Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, China.
Department of Infection, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, China.
Comput Math Methods Med. 2023 Feb 16;2023:1318817. doi: 10.1155/2023/1318817. eCollection 2023.
Acute kidney injury (AKI) is a clinical syndrome caused by various reasons that results in the rapid decline of renal function in a short period of time. Severe AKI can lead to multiple organ dysfunction syndrome. Circular RNA HIPK3 (circHIPK3) derived from the gene is involved in multiple inflammatory processes. The present research was performed to explore the function of circHIPK3 on AKI. The AKI model was established by ischemia/reperfusion (I/R) in C57BL/6 mice or hypoxia/reoxygenation (H/R) in HK-2 cells. The function and mechanism of circHIPK3 on AKI were explored via biochemical index measurement; hematoxylin and eosin (HE) staining; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT); flow cytometry; enzyme-linked immunosorbent assay (ELISA); western blot; quantitative real-time polymerase chain reaction (RT-qPCR); detection of reactive oxygen species (ROS) and adenosine triphosphate (ATP); and luciferase reporter assays. circHIPK3 was upregulated in kidney tissues of I/R-induced mice and in H/R-treated HK-2 cells, while the microRNA- (miR-) 93-5p level was decreased in H/R-stimulated HK-2 cells. Furthermore, circHIPK3 silencing or miR-93-5p overexpression could reduce the level of proinflammatory factors and oxidative stress and recover the cell viability in H/R-stimulated HK-2 cells. Meanwhile, the luciferase assay showed that Krüppel-like transcription factor 9 (KLF9) was the downstream target of miR-93-5p. Forced expression of KLF9 blocked the function of miR-93-5p on H/R-treated HK-2 cells. Knockdown of circHIPK3 improved the renal function and reduced the apoptosis level . In conclusion, circHIPK3 knockdown alleviated oxidative stress and apoptosis and inhibited inflammation in AKI via miR-93-5p-mediated downregulation of the KLF9 signal pathway.
急性肾损伤 (AKI) 是由多种原因引起的临床综合征，导致肾功能在短时间内迅速下降。严重的 AKI 可导致多器官功能障碍综合征。来源于基因的环状 RNA HIPK3 (circHIPK3) 参与了多种炎症过程。本研究旨在探讨 circHIPK3 对 AKI 的作用。通过缺血/再灌注 (I/R) 在 C57BL/6 小鼠或缺氧/复氧 (H/R) 在 HK-2 细胞中建立 AKI 模型。通过生化指标测定、苏木精和伊红 (HE) 染色、3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐 (MTT)、流式细胞术、酶联免疫吸附测定 (ELISA)、western blot、实时定量聚合酶链反应 (RT-qPCR)、活性氧 (ROS) 和三磷酸腺苷 (ATP) 检测、荧光素酶报告基因实验，探讨 circHIPK3 对 AKI 的作用及机制。在 I/R 诱导的小鼠肾脏组织和 H/R 处理的 HK-2 细胞中，circHIPK3 上调，而在 H/R 刺激的 HK-2 细胞中，miR-93-5p 水平下降。此外，circHIPK3 沉默或 miR-93-5p 过表达可降低 H/R 刺激的 HK-2 细胞中促炎因子和氧化应激水平，恢复细胞活力。同时，荧光素酶报告基因实验表明，Krüppel 样转录因子 9 (KLF9) 是 miR-93-5p 的下游靶基因。KLF9 的强制表达阻断了 miR-93-5p 对 H/R 处理的 HK-2 细胞的作用。circHIPK3 的敲低改善了肾功能，降低了。总之，circHIPK3 的敲低通过 miR-93-5p 介导的 KLF9 信号通路下调减轻 AKI 中的氧化应激和细胞凋亡，抑制炎症。
