Department of Nephrology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China,
Department of Urology, The Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai, China.
Kidney Blood Press Res. 2020;45(2):209-221. doi: 10.1159/000505154. Epub 2020 Feb 18.
BACKGROUND/AIMS: The purpose of this study is to analyze the expression and biological function of lncRNA ANRIL, microRNA-199a, TLR4, and nuclear factor-kappa B (NF-κB) in acute renal injury (AKI) induced by lipopolysaccharide (LPS).
The levels of ANRIL and microRNA-199a in mouse cells and kidneys were detected by quantitative-polymerase chain reaction. Western blot analysis was used for the NF-κB pathway protein. MTT assay was used for cell viability. Enzyme-linked immunosorbent assay was used for the secretion of inflammatory factors in mouse kidney tissue. Apoptosis was measured by flow cytometry and Western blotting. The potential binding region between ANRIL and miR-199a was verified by luciferase reporter assay.
The upregulation of ANRIL can reduce the expression of microRNA-199a and increases the number of apoptotic cells. The expression levels of ANRIL in LPS-induced AKI mice and LPS-treated HK2 cells were upregulated compared with the control group. Overexpression of ANRIL increased apoptosis and promoted TLR4 (Toll-like receptor 4), NF-κB phosphorylation, and downstream transcription factor production.
ANRIL/NF-κB pathway in LPS-induced apoptosis provided theoretical guidance for ANRIL in the treatment of AKI.
背景/目的：本研究旨在分析长链非编码 RNA（lncRNA）ANRIL、微小 RNA-199a、Toll 样受体 4（TLR4）和核因子-κB（NF-κB）在脂多糖（LPS）诱导的急性肾损伤（AKI）中的表达和生物学功能。
通过定量聚合酶链反应检测小鼠细胞和肾脏中 ANRIL 和 microRNA-199a 的水平。采用 Western blot 分析 NF-κB 通路蛋白。MTT 法检测细胞活力。酶联免疫吸附试验检测小鼠肾组织中炎症因子的分泌。通过流式细胞术和 Western blot 检测细胞凋亡。通过荧光素酶报告试验验证 ANRIL 与 miR-199a 之间的潜在结合区域。
上调 ANRIL 可降低 microRNA-199a 的表达并增加凋亡细胞数量。与对照组相比，LPS 诱导的 AKI 小鼠和 LPS 处理的 HK2 细胞中 ANRIL 的表达上调。过表达 ANRIL 增加了细胞凋亡，并促进了 TLR4、NF-κB 磷酸化和下游转录因子的产生。
LPS 诱导的凋亡中 ANRIL/NF-κB 通路为 ANRIL 在 AKI 治疗中的应用提供了理论指导。
