长链非编码 RNA SNHG14 通过调控 miR-124-3p/MMP2 轴缓解缺血/再灌注诱导的急性肾损伤。

Silence of Long Noncoding RNA SNHG14 Alleviates Ischemia/Reperfusion-Induced Acute Kidney Injury by Regulating miR-124-3p/MMP2 Axis.

机构信息

Department of Emergency, The First Affiliated Hospital of Hebei North University, No. 12, Changqing Road, Zhangjiakou City, Hebei Province 075000, China.

出版信息

Biomed Res Int. 2021 Jan 4;2021:8884438. doi: 10.1155/2021/8884438. eCollection 2021.

PURPOSE

Ample evidence has proved that lncRNAs are pivotal regulators in acute kidney injury (AKI). Here, we focus on the role and mechanism of lncRNA SNHG14 in ischemia/reperfusion- (I/R-) caused AKI.

METHODS

I/R and hypoxia/reoxygenation (H/R) were applied to induce rats and HK-2 cells to establish AKI models in vivo and in vitro. Relative expression of SNHG14, miR-124-3p, and MMP2 was determined by qRT-PCR. HE staining was used to evaluate pathological changes in renal tissues, and acute tubular necrosis (ATN) score was calculated. Renal function was evaluated by measuring serum creatinine content and blood urea nitrogen content. Levels of IL-1, IL-6, and TNF- were measured by ELISA. Cell viability was examined by MTT assay. Oxidative stress was assessed by measuring SOD, MDA, and ROS levels. The target of SNHG14 or miR-124-3p was verified by DLR assay. Protein expression of MMP2 was examined by western blot.

RESULTS

SNHG14 was boosted in renal tissues of I/R-stimulated rats and H/R-induced HK-2 cells, while miR-124-3p was diminished in H/R-stimulated HK-2 cells. Si-SNHG14 or miR-124-3p mimics repressed inflammation and oxidative stress and enhanced cell viability in H/R-stimulated HK-2 cells. Sh-SNHG14 mitigated I/R-induced AKI in rats. MiR-124-3p was targeted by SNHG14, and MMP2 was targeted by miR-124-3p. Inhibition of miR-124-3p or upregulation of MMP2 reversed inhibitory effects of SNHG14 silence on inflammation and oxidative stress as well as the promoting effect of SNHG14 silence on cell viability in H/R-induced HK-2 cells.

CONCLUSION

Knockdown of SNHG14 alleviated I/R-induced AKI by miR-124-3p-mediated downregulation of MMP2.

目的

大量证据表明，lncRNAs 是急性肾损伤（AKI）的关键调节因子。在这里，我们专注于 lncRNA SNHG14 在缺血/再灌注（I/R）引起的 AKI 中的作用和机制。

方法

应用 I/R 和缺氧/复氧（H/R）在体内和体外诱导大鼠和 HK-2 细胞建立 AKI 模型。通过 qRT-PCR 测定 SNHG14、miR-124-3p 和 MMP2 的相对表达。HE 染色用于评估肾组织的病理变化，并计算急性肾小管坏死（ATN）评分。通过测量血清肌酐含量和血尿素氮含量评估肾功能。通过 ELISA 测定 IL-1、IL-6 和 TNF-的水平。通过 MTT 测定法检查细胞活力。通过测量 SOD、MDA 和 ROS 水平评估氧化应激。通过 DLR 测定验证 SNHG14 或 miR-124-3p 的靶标。通过 Western blot 检查 MMP2 蛋白表达。

结果

SNHG14 在 I/R 刺激的大鼠肾组织和 H/R 诱导的 HK-2 细胞中上调，而 miR-124-3p 在 H/R 刺激的 HK-2 细胞中下调。Si-SNHG14 或 miR-124-3p 模拟物抑制 H/R 刺激的 HK-2 细胞中的炎症和氧化应激并增强细胞活力。Sh-SNHG14 减轻了大鼠的 I/R 诱导的 AKI。miR-124-3p 是 SNHG14 的靶标，而 MMP2 是 miR-124-3p 的靶标。抑制 miR-124-3p 或上调 MMP2 逆转了 SNHG14 沉默对 H/R 诱导的 HK-2 细胞中炎症和氧化应激的抑制作用以及 SNHG14 沉默对细胞活力的促进作用。