Hsa Circ 001839 通过 miR-432-3p 促进肾缺血再灌注损伤中的 NLRP3 炎症反应。

Hsa Circ 001839 Promoted Inflammation in Renal Ischemia-Reperfusion Injury Through NLRP3 by miR-432-3p.

机构信息

Department of Urology, Luzhou People's Hospital, Luzhou, China.

Department of Epidemiology and Statistics, School of Public Health, Southwest Medical University, Luzhou, China.

出版信息

Nephron. 2021;145(5):540-552. doi: 10.1159/000515279. Epub 2021 May 11.

DOI:10.1159/000515279

PMID:33975327

Abstract

BACKGROUND

In recent years, increasing discovery of the extremely important regulatory effects of circular RNAs on biological development, angiogenesis, tumor genesis, and development, as well as stem cell proliferation and differentiation has provided new opportunities for investigating regulation mechanism in angiogenesis.

OBJECTIVES

This study explored the expression of circ 001839 in renal ischemia-reperfusion injury (RI-RI) rats and whether its upstream microRNA-432-3p (miR-432-3p) affects inflammation in both RI-RI rats and NRK52E cells.

METHODS

Rat model of RI-RI was made, and circ 001839 was identified by the gene-chip analysis in RI-RI rats. Expression of circ 001839 and miR-432-3p was measured by reverse transcription-quantitative polymerase chain reaction, protein expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, interferon (IFN)-γ, IL-6, and IL-18 in rat serum and cell supernatant was determined by ELISA, and the expression of NOD-like receptor 3 (NLRP3) and other gap-associated proteins in NRK52E cells was evaluated by Western blot analysis. Next, to verify the regulatory relationship between circ 001839 and miR-432-3p, 2 luciferase reporters were constructed.

RESULTS

Circ 001839 expression of RI-RI rats and NRK52E cells was significantly upregulated, compared with the control group. Circ 001839 overexpression significantly increased inflammation through promoting TNF-α, IFN-γ, and IL-6 expression levels in NRK52E cells. Overexpression of miR-432-3p significantly promoted inflammation in NRK52E cells via induction of NLRP3. Moreover, miR-432-3p decreased the effects of circ 001839-induced inflammation in NRK52E cells.

CONCLUSIONS

These findings suggested that circ 001839 promoted inflammation in RI-RI through NLRP3 by miR-432-3p.

摘要

背景

近年来，环状 RNA 对生物发育、血管生成、肿瘤发生和发展以及干细胞增殖和分化的极其重要的调控作用的不断发现，为研究血管生成的调控机制提供了新的机会。

目的

本研究探讨了 circ 001839 在肾缺血再灌注损伤（RI-RI）大鼠中的表达及其上游 microRNA-432-3p（miR-432-3p）是否影响 RI-RI 大鼠和 NRK52E 细胞的炎症。

方法

建立 RI-RI 大鼠模型，通过基因芯片分析鉴定 RI-RI 大鼠中的 circ 001839。采用逆转录定量聚合酶链反应测定 circ 001839 和 miR-432-3p 的表达，酶联免疫吸附法测定大鼠血清和细胞上清液中肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、干扰素（IFN）-γ、IL-6 和 IL-18 的蛋白表达，Western blot 分析 NRK52E 细胞中 NOD 样受体 3（NLRP3）和其他间隙连接蛋白的表达。接下来，为了验证 circ 001839 与 miR-432-3p 的调控关系，构建了 2 个荧光素酶报告基因。

结果

与对照组相比，RI-RI 大鼠和 NRK52E 细胞的 circ 001839 表达明显上调。circ 001839 过表达通过促进 NRK52E 细胞中 TNF-α、IFN-γ 和 IL-6 的表达水平显著增加炎症。miR-432-3p 的过表达通过诱导 NLRP3 显著促进 NRK52E 细胞的炎症。此外，miR-432-3p 降低了 circ 001839 在 NRK52E 细胞中诱导的炎症作用。