The Prognostic Value of LncRNA NNT-AS1 in CRRT-Treated Patients and Its Regulatory Mechanism in Acute Kidney Injury.

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Nephrology (Carlton, Vic.) Pub Date : 2025-10-01 DOI:10.1111/nep.70120

Shiming Zhang, Xuechun Wang, Ping Ni, Peng Huang, Lili Liang

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Abstract

Aim: We investigated the prognostic value of lncRNA NNT-AS1 in patients undergoing CRRT and its regulatory mechanism in acute kidney injury.

Methods: First, we detected serum levels of NNT-AS1 by RT-qPCR in patients before and after treatment with CRRT. Next, the survival probability between NNT-AS1 levels and prognosis of CRRT-treated patients was explored with Kaplan-Meier (K-M) curve analysis. The potential risk factors were evaluated via Cox regression analysis. Moreover, LPS-induced HK-2 cells were employed to establish an in vitro cellular experimental model. Cell viability and apoptosis rate were measured by Cell Counting Kit-8 (CCK-8) and flow cytometry. The relative mRNA levels of MDA and ROS were detected via RT-qPCR, and the concentration of tumour necrosis factor-α (TNF-α), interleukins-1β (IL-1β), neutrophil gelatinase-associated lipocalin (NGAL) and kidney injury molecule 1 (KIM-1) was detected by ELISA assay. Finally, a dual-luciferase reporter assay was utilised to verify the targeted binding of NNT-AS1 to miR-582-5p.

Results: Serum NNT-AS1 levels were significantly reduced following CRRT, and patients with high levels of NNT-AS1 had lower survival probability. NNT-AS1 is a potential risk factor for poor outcomes in patients treated with CRRT. In LPS-induced HK-2 cells, NNT-AS1 knockdown increased cell viability and reduced the apoptosis rate. Also, NNT-AS1 knockdown suppressed the expression of oxidative stress markers, inflammatory factors and markers of renal tubular injury. NNT-AS1 targeted miR-582-5p by dual-luciferase reporter assay.

Conclusion: LncRNA NNT-AS1 can serve as a marker of poor prognosis for CRRT treatment as well as participate in AKI by targeting miR-582-5p.

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LncRNA NNT-AS1在crrt治疗患者中的预后价值及其在急性肾损伤中的调节机制

目的：探讨lncRNA NNT-AS1在CRRT患者中的预后价值及其在急性肾损伤中的调控机制。方法：首先，采用RT-qPCR方法检测CRRT治疗前后患者血清NNT-AS1水平。接下来，采用Kaplan-Meier （K-M）曲线分析探讨NNT-AS1水平与crrt治疗患者预后之间的生存概率。通过Cox回归分析评估潜在危险因素。采用lps诱导的HK-2细胞建立体外细胞实验模型。采用细胞计数试剂盒-8 （CCK-8）和流式细胞术检测细胞活力和凋亡率。RT-qPCR检测MDA和ROS的相对mRNA水平，ELISA法检测肿瘤坏死因子-α (TNF-α)、白细胞介素-1β (IL-1β)、中性粒细胞明胶酶相关脂钙素（NGAL）和肾损伤分子1 （KIM-1）的浓度。最后，采用双荧光素酶报告试验验证NNT-AS1与miR-582-5p的靶向结合。结果：CRRT后血清NNT-AS1水平显著降低，且NNT-AS1水平高的患者生存几率较低。NNT-AS1是CRRT治疗患者预后不良的潜在危险因素。在lps诱导的HK-2细胞中，NNT-AS1敲低可提高细胞活力，降低细胞凋亡率。此外，NNT-AS1敲低可抑制氧化应激标志物、炎症因子和肾小管损伤标志物的表达。通过双荧光素酶报告基因检测NNT-AS1靶向miR-582-5p。结论：LncRNA NNT-AS1可以作为CRRT治疗不良预后的标志物，并通过靶向miR-582-5p参与AKI。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Nephrology (Carlton, Vic.)

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