NEAT1 通过吸附 miR-22-3p 加重脓毒症诱导的急性肾损伤。
NEAT1 aggravates sepsis-induced acute kidney injury by sponging miR-22-3p.
作者信息
Feng Yawei, Liu Jun, Wu Ranliang, Yang Peng, Ye Zhiqiang, Song Furong
机构信息
Department of Anesthesiology, The Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
Department of Anesthesiology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong, No. 58, Zhongshan Erlu, Guangzhou 510080, China.
出版信息
Open Med (Wars). 2020 Apr 20;15(1):333-342. doi: 10.1515/med-2020-0401. eCollection 2020.
BACKGROUND AND AIM
Acute kidney injury (AKI) is a common complication of sepsis. Long noncoding RNA nuclear-enriched abundant transcript 1 (NEAT1) plays a vital role in various diseases, including AKI. This study aimed to investigate the function and mechanism of NEAT1 in sepsis-induced AKI.
MATERIALS AND METHODS
A septic AKI model was established by treating HK-2 cells with lipopolysaccharide (LPS). The levels of NEAT1 and miR-22-3p were measured by quantitative real-time PCR. Cell apoptosis was assessed by flow cytometry. The levels of apoptosis-related protein and autophagy-related factors were examined by the western blot assay. An enzyme-linked immunosorbent assay was used to calculate the contents of inflammatory factors. The interaction between NEAT1 and miR-22-3p was validated by dual-luciferase reporter assay, RNA immunoprecipitation assay, and RNA pull-down assay. The levels of nuclear factor (NF)-κB pathway-related proteins were evaluated by the western blot assay.
RESULTS
NEAT1 was upregulated, while miR-22-3p was downregulated in patients with sepsis and in LPS-stimulated HK-2 cells. LPS treatment triggered cell apoptosis, autophagy, and inflammatory response in HK-2 cells. NEAT1 knockdown attenuated LPS-induced cell injury. NEAT1 modulated LPS-triggered cell injury by targeting miR-22-3p. Furthermore, NEAT1 regulated the NF-κB pathway by modulating miR-22-3p.
CONCLUSION
Depletion of NEAT1 alleviated sepsis-induced AKI via regulating the miR-22-3p/NF-κB pathway.
背景与目的
急性肾损伤（AKI）是脓毒症的常见并发症。长链非编码RNA核富集丰富转录本1（NEAT1）在包括AKI在内的多种疾病中发挥着至关重要的作用。本研究旨在探讨NEAT1在脓毒症诱导的AKI中的作用及机制。
材料与方法
通过用脂多糖（LPS）处理HK-2细胞建立脓毒症AKI模型。采用定量实时PCR检测NEAT1和miR-22-3p的水平。通过流式细胞术评估细胞凋亡。采用蛋白质印迹法检测凋亡相关蛋白和自噬相关因子的水平。采用酶联免疫吸附测定法计算炎症因子的含量。通过双荧光素酶报告基因测定、RNA免疫沉淀测定和RNA下拉测定验证NEAT1与miR-22-3p之间的相互作用。采用蛋白质印迹法评估核因子（NF）-κB通路相关蛋白的水平。
结果
脓毒症患者和LPS刺激的HK-2细胞中NEAT1上调，而miR-22-3p下调。LPS处理引发HK-2细胞凋亡、自噬和炎症反应。敲低NEAT1可减轻LPS诱导的细胞损伤。NEAT1通过靶向miR-22-3p调节LPS触发的细胞损伤。此外，NEAT1通过调节miR-22-3p调控NF-κB通路。
结论
NEAT1的缺失通过调节miR-22-3p/NF-κB通路减轻脓毒症诱导的AKI。
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