Department of Emergency, Affiliated Hospital of North Sichuan Medical College, Nanchong City, China.
Department of Critical care, Affiliated Hospital of North Sichuan Medical College, Nanchong City, China.
Shock. 2024 Jan 1;61(1):41-48. doi: 10.1097/SHK.0000000000002229. Epub 2023 Oct 16.
Background: Sepsis is a life-threatening systemic inflammatory disease that can cause many diseases, including acute kidney injury (AKI). Increasing evidence showed that a variety of circular RNAs were considered to be involved in the development of the disease. In this study, we aimed to elucidate the role and potential mechanism of circUSP42 in sepsis-induced AKI. Methods: HK2 cells were treated with lipopolysaccharide (LPS) to establish septic AKI cell model. The expression levels of circUSP42, microRNA-182-5p (miR-182-5p), and DUSP1 in LPS-treated HK2 cells were measured by quantitative real-time polymerase chain reaction or Western blot. Functional experiments were performed by using Cell Counting Kit-8, 5-ethynyl-2'-deoxyuridine staining, flow cytometry, oxidative stress assay, and enzyme-linked immunosorbent assay. The potential target binding site between miR-182-5p and CircUSP42 or DUSP1 was verified by dual-luciferase reporter and RNA immunoprecipitation assays. Results: CircUSP42 and DUSP1 were downregulated in serum samples from patients with AKI and LPS-treated HK2 cells, while miR-182-5p was upregulated. Functionally, overexpression of CircUSP42 promoted cell proliferation and inhibited apoptosis, inflammation, and oxidative stress in LPS-triggered HK2 cells. Further mechanism analysis showed that miR-182-5p had potential binding sites with circUSP42 and DUSP1, and circUSP42 regulated LPS-induced cell damage by targeting miR-182-5p. At the same time, miR-182-5p knockdown inhibited LPS-treated HK2 cell damage by regulating DUSP1. In addition, circUSP42 induced DUSP1 expression via sponging miR-182-5p to ameliorate LPS-induced HK2 cell damage. Conclusion : Our results showed that circUSP42 overexpression might attenuate LPS-induced HK2 cell injury by regulating miR-182-5p/DUSP1 axis. This might provide therapeutic strategy for the treatment of sepsis.
脓毒症是一种危及生命的全身性炎症性疾病，可导致多种疾病，包括急性肾损伤（AKI）。越来越多的证据表明，多种环状 RNA 被认为参与了疾病的发展。在本研究中，我们旨在阐明 circUSP42 在脓毒症诱导的 AKI 中的作用和潜在机制。
用脂多糖（LPS）处理 HK2 细胞建立脓毒症 AKI 细胞模型。用实时定量聚合酶链反应或 Western blot 检测 LPS 处理的 HK2 细胞中 circUSP42、microRNA-182-5p（miR-182-5p）和 DUSP1 的表达水平。通过细胞计数试剂盒-8、5-乙炔基-2'-脱氧尿苷染色、流式细胞术、氧化应激测定和酶联免疫吸附测定进行功能实验。通过双荧光素酶报告和 RNA 免疫沉淀测定验证 miR-182-5p 与 circUSP42 或 DUSP1 之间的潜在靶标结合位点。
AKI 患者血清样本和 LPS 处理的 HK2 细胞中 circUSP42 和 DUSP1 下调，而 miR-182-5p 上调。功能上，circUSP42 的过表达促进了 LPS 触发的 HK2 细胞的增殖，并抑制了细胞凋亡、炎症和氧化应激。进一步的机制分析表明，miR-182-5p 与 circUSP42 和 DUSP1 具有潜在的结合位点，circUSP42 通过靶向 miR-182-5p 调节 LPS 诱导的细胞损伤。同时，miR-182-5p 敲低通过调节 DUSP1 抑制 LPS 处理的 HK2 细胞损伤。此外，circUSP42 通过海绵吸附 miR-182-5p 诱导 DUSP1 表达，从而减轻 LPS 诱导的 HK2 细胞损伤。
我们的结果表明，circUSP42 的过表达可能通过调节 miR-182-5p/DUSP1 轴来减轻 LPS 诱导的 HK2 细胞损伤。这可能为脓毒症的治疗提供一种策略。
