Knockdown of long non-coding RNA SBF2-AS1 inhibits calcium oxalate-induced HK-2 cell injury by regulating the miR-302e/NLRP3 pathway.

IF 2 2区医学 Q2 UROLOGY & NEPHROLOGY

Urolithiasis Pub Date : 2024-08-06 DOI:10.1007/s00240-024-01606-y

Hanliang Zhu, Yiwen Chen, Yuantang Zhong, Xiangwei Xie, Xiangjian Zeng, Wangdong Deng

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Abstract

Long non-coding ribose nucleic acids (lncRNAs) have been implicated in the development of nephrolithiasis. The study aims to investigate the interplay of lncRNA SBF2-AS1 (SETbinding factor 2 antisense RNA 1) and NLR family pyrin domain containing 3 (NLRP3) in regulating the calcium oxalate monohydrate (COM)-induced human kidney HK-2 cell injury. HK-2 cells were treated with COM (100 µg/mL) to create a cellular model of kidney injury. Gene and protein expression was assessed by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blot. Proliferation and apoptosis rates, as well as levels of malondialdehyde (MDA), lactate dehydrogenase (LDH), superoxide dismutase (SOD), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 were measured. Additionally, potential miRNAs interacting with SBF2-AS1 and NLRP3 were predicted utilizing the starBase and TargetScan databases. The interference of SBF2-AS1 resulted in increased cell proliferation and SOD levels in HK-2 cells after COM induction. SBF2-AS1 silencing also reduced COM-induced cell death and inflammatory cytokine production by down-regulating NLRP3 protein expression. Conversely, forced upregulation of NLRP3 abrogated the effect of SBF2-AS1 interference. Notably, SBF2-AS1 interference on COM-induced oxidative stress and COM-induced cellular damage was rescued by antioxidant, indicating the involvement of oxidative burden in COM-induced damage. miR-302e acted as a mediator miRNA linking the functional association of SBF2-AS1 and NLRP3. Silencing SBF2-AS1 promoted miR-302e level and miR-302e reduced NLRP3 expression in HK-2 cells to protect against COM-induced damage. In summary, these findings suggest that downregulation of lncRNA SBF2-AS1 can potentially protect HK-2 cells from COM-induced injury by modulating the miR-302e/NLRP3 pathway.

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敲除长非编码 RNA SBF2-AS1 通过调节 miR-302e/NLRP3 通路抑制草酸钙诱导的 HK-2 细胞损伤

长非编码核糖核酸（lncRNA）被认为与肾炎的发生有关。本研究旨在探讨lncRNA SBF2-AS1 （SET结合因子2反义RNA 1）和NLR家族含吡咯啉结构域3（NLRP3）在调控一水草酸钙（COM）诱导的人肾HK-2细胞损伤中的相互作用。用COM（100 µg/mL）处理HK-2细胞，建立肾损伤细胞模型。基因和蛋白质表达通过定量反转录聚合酶链反应（qRT-PCR）和Western印迹进行评估。还测定了细胞的增殖率和凋亡率，以及丙二醛（MDA）、乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和IL-6的水平。此外，还利用 starBase 和 TargetScan 数据库预测了与 SBF2-AS1 和 NLRP3 相互作用的潜在 miRNA。干扰SBF2-AS1会导致HK-2细胞在COM诱导后细胞增殖和SOD水平升高。通过下调 NLRP3 蛋白的表达，SBF2-AS1 的沉默也减少了 COM 诱导的细胞死亡和炎性细胞因子的产生。相反，强制上调 NLRP3 会削弱 SBF2-AS1 干扰的效果。值得注意的是，SBF2-AS1干扰对COM诱导的氧化应激和COM诱导的细胞损伤有抗氧化作用，表明氧化负担参与了COM诱导的损伤。沉默 SBF2-AS1可促进 miR-302e 的水平，而 miR-302e 可降低 HK-2 细胞中 NLRP3 的表达，从而保护细胞免受 COM 诱导的损伤。综上所述，这些研究结果表明，下调lncRNA SBF2-AS1可通过调节miR-302e/NLRP3通路保护HK-2细胞免受COM诱导的损伤。

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来源期刊

Urolithiasis UROLOGY & NEPHROLOGY-

CiteScore

4.50

自引率

6.50%

发文量

期刊介绍： Official Journal of the International Urolithiasis Society The journal aims to publish original articles in the fields of clinical and experimental investigation only within the sphere of urolithiasis and its related areas of research. The journal covers all aspects of urolithiasis research including the diagnosis, epidemiology, pathogenesis, genetics, clinical biochemistry, open and non-invasive surgical intervention, nephrological investigation, chemistry and prophylaxis of the disorder. The Editor welcomes contributions on topics of interest to urologists, nephrologists, radiologists, clinical biochemists, epidemiologists, nutritionists, basic scientists and nurses working in that field. Contributions may be submitted as full-length articles or as rapid communications in the form of Letters to the Editor. Articles should be original and should contain important new findings from carefully conducted studies designed to produce statistically significant data. Please note that we no longer publish articles classified as Case Reports. Editorials and review articles may be published by invitation from the Editorial Board. All submissions are peer-reviewed. Through an electronic system for the submission and review of manuscripts, the Editor and Associate Editors aim to make publication accessible as quickly as possible to a large number of readers throughout the world.