长链非编码 RNA NKILA 敲低通过调控 miR-140-5p/CLDN2 轴促进脂多糖诱导的脓毒症模型中的细胞活力，抑制细胞凋亡、自噬和炎症。
LncRNA NKILA knockdown promotes cell viability and represses cell apoptosis, autophagy and inflammation in lipopolysaccharide-induced sepsis model by regulating miR-140-5p/CLDN2 axis.
机构信息
Department of Emergency Internal Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai City, China.
Department of Emergency Internal Medicine, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai City, China.
出版信息
Biochem Biophys Res Commun. 2021 Jun 25;559:8-14. doi: 10.1016/j.bbrc.2021.04.074. Epub 2021 Apr 28.
BACKGROUND
Long non-coding RNAs (lncRNAs) play vital roles in human diseases, including sepsis-induced acute kidney injury (AKI). Here, we aimed to investigate the functions of lncRNA NKILA in sepsis-engendered AKI.
METHODS
HK2 cells stimulated with LPS were used to mimic sepsis-induced AKI in vitro. qRT-PCR was conducted for lncRNA NKILA and miR-140-5p levels. Cell Counting Kit-8 (CCK-8) assay and flow cytometry analysis were employed to analyze cell viability and apoptosis. Western blot assay was utilized to measured protein levels. ELISA kits were used to examine the concentrations of IL-6, IL-1β and TNF-α. Dual-luciferase reporter assay was utilized to analyze the relationships among lncRNA NKILA, miR-140-5p and claudin 2 (CLDN2).
RESULTS
LPS restrained HK2 cell viability and accelerated cell apoptosis and autophagy. LncRNA NKILA was increased in LPS-treated HK2 cells. LncRNA NKILA silencing reversed the promotional influence of LPS on cell progression in HK2 cells. miR-140-5p inhibition ameliorated lncRNA NKILA knockdown-mediated cell injury in LPS-mediated HK2 cells. CLDN2 was the target of miR-140-5p. MiR-140-5p elevation promoted cell viability and suppressed cell apoptosis, autophagy and inflammation in LPS-induced HK2 cells, with CLDN2 elevation overturned the effects.
CONCLUSION
LncRNA NKILA silencing protected HK2 cells from LPS-induced impairments by reducing CLDN2 through sponging miR-140-5p.
背景
长链非编码 RNA（lncRNA）在包括脓毒症诱导的急性肾损伤（AKI）在内的人类疾病中发挥着重要作用。在这里，我们旨在研究 lncRNA NKILA 在脓毒症引起的 AKI 中的作用。
方法
用 LPS 刺激 HK2 细胞在体外模拟脓毒症诱导的 AKI。用 qRT-PCR 检测 lncRNA NKILA 和 miR-140-5p 的水平。用细胞计数试剂盒-8（CCK-8）检测和流式细胞术分析检测细胞活力和细胞凋亡。用 Western blot 检测蛋白水平。用 ELISA 试剂盒检测 IL-6、IL-1β 和 TNF-α的浓度。用双荧光素酶报告基因实验分析 lncRNA NKILA、miR-140-5p 和紧密连接蛋白 2（CLDN2）之间的关系。
结果
LPS 抑制 HK2 细胞活力，加速细胞凋亡和自噬。LPS 处理的 HK2 细胞中 lncRNA NKILA 增加。LncRNA NKILA 沉默逆转了 LPS 对 HK2 细胞增殖的促进作用。miR-140-5p 抑制改善了 LPS 介导的 HK2 细胞中 lncRNA NKILA 敲低介导的细胞损伤。CLDN2 是 miR-140-5p 的靶标。miR-140-5p 升高促进了 LPS 诱导的 HK2 细胞的细胞活力，并抑制了细胞凋亡、自噬和炎症，而 CLDN2 的升高则逆转了这些作用。
结论
LncRNA NKILA 沉默通过海绵吸附 miR-140-5p 减少 CLDN2，从而保护 HK2 细胞免受 LPS 诱导的损伤。
Zotero 插件