环状 RNA HIPK3 通过 miRNA-124-3p/miRNA-148b-3p 调控炎症反应和细胞衰老在白念珠菌诱导的脓毒症急性肾损伤中的作用机制
Mechanism of circHIPK3-miRNA-124-3p/miRNA-148b-3p-Mediated Inflammatory Responses and Cell Senescence in Candida albicans-Induced Septic Acute Kidney Injury.
机构信息
Department of Critical Care Medicine, The Fifth Affiliated Hospital (Zhuhai) of Zunyi Medical University, Zhuhai, China.
Department of Emergency, The Fifth Affiliated Hospital (Zhuhai) of Zunyi Medical University, Zhuhai, China.
出版信息
Gerontology. 2022;68(10):1145-1165. doi: 10.1159/000523910. Epub 2022 May 16.
INTRODUCTION
Sepsis is a life-threatening inflammatory state that can result in septic acute kidney injury (SAKI). Circular RNAs (circRNAs) are implicated in various inflammatory diseases including SAKI. This study investigated the effect of circHIPK3 on inflammatory responses and cell senescence in Candida albicans-induced SAKI.
METHODS
circHIPK3 expression and inflammatory factors in the serum of SAKI patients and healthy volunteers were detected. The murine and cell models of SAKI were established by C. albicans and lipopolysaccharide induction, respectively. The effect of circHIPK3 on SAKI inflammatory responses and cell senescence was measured using ELISA, SA-β-gal staining, CCK-8, RT-qPCR, and Western blot. The binding relationships among circHIPK3, miR-124-3p, or miR-148b-3p and KLF6 or DNMT1/3a were confirmed. The binding of KLF6 and NLRP3 was determined, and the methylation level of the Klotho promoter was detected. Functional rescue experiments were performed to verify the effect of miR-124-3p or miR-148b-3p on SAKI.
RESULTS
circHIPK3 was highly expressed in SAKI. circHIPK3 silencing alleviated kidney injury in SAKI mice and enhanced SAKI cell viability by alleviating inflammatory responses and cell senescence. Mechanically, circHIPK3 upregulated KLF6 expression by competitively binding to miR-124-3p, thereby promoting the binding of KLF6 and NLRP3, activating NLRP3/caspase-1-mediated pyroptosis, and eventually aggravating SAKI inflammatory responses. circHIPK3 upregulated DNMT1/3a expression by competitively binding to miR-148b-3p, thus elevating the methylation level of Klotho promoter and accelerating SAKI cell senescence. Downregulation of miR-124-3p or miR-148b-3p attenuated the protective effect of circHIPK3 silencing on SAKI.
CONCLUSION
circHIPK3 aggravated SAKI inflammatory responses via miR-124-3p/KLF6 and accelerated SAKI cell senescence via miR-148b-3p/DNMT1/3a.
简介
败血症是一种危及生命的炎症状态，可导致败血症性急性肾损伤（SAKI）。环状 RNA（circRNA）与包括 SAKI 在内的各种炎症性疾病有关。本研究探讨了 circHIPK3 对白色念珠菌诱导的 SAKI 中炎症反应和细胞衰老的影响。
方法
检测 SAKI 患者和健康志愿者血清中 circHIPK3 的表达和炎症因子。分别通过白色念珠菌和脂多糖诱导建立 SAKI 小鼠和细胞模型。通过 ELISA、SA-β-半乳糖染色、CCK-8、RT-qPCR 和 Western blot 检测 circHIPK3 对 SAKI 炎症反应和细胞衰老的影响。验证 circHIPK3 与 miR-124-3p 或 miR-148b-3p 与 KLF6 或 DNMT1/3a 之间的结合关系。确定 KLF6 和 NLRP3 的结合情况，并检测 Klotho 启动子的甲基化水平。进行功能挽救实验以验证 miR-124-3p 或 miR-148b-3p 对 SAKI 的影响。
结果
circHIPK3 在 SAKI 中高表达。circHIPK3 沉默通过减轻炎症反应和细胞衰老减轻 SAKI 小鼠的肾损伤并增强 SAKI 细胞活力。机制上，circHIPK3 通过竞争性结合 miR-124-3p 上调 KLF6 表达，从而促进 KLF6 和 NLRP3 的结合，激活 NLRP3/caspase-1 介导的细胞焦亡，最终加重 SAKI 炎症反应。circHIPK3 通过竞争性结合 miR-148b-3p 上调 DNMT1/3a 表达，从而提高 Klotho 启动子的甲基化水平并加速 SAKI 细胞衰老。下调 miR-124-3p 或 miR-148b-3p 可减弱 circHIPK3 沉默对 SAKI 的保护作用。
结论
circHIPK3 通过 miR-124-3p/KLF6 加重 SAKI 炎症反应，并通过 miR-148b-3p/DNMT1/3a 加速 SAKI 细胞衰老。
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