Urology, Longgang District Central Hospital of Shenzhen, 6082 Longgang Avenue, Longgang District, Shenzhen City, 518116, Guangdong Province, China.
Urolithiasis. 2024 Aug 6;52(1):113. doi: 10.1007/s00240-024-01606-y.
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.
长链非编码核糖核酸（lncRNA）已被认为与肾结石的发生有关。本研究旨在探讨 lncRNA SBF2-AS1（SET 结合因子 2 反义 RNA 1）和富含 N 端亮氨酸重复序列的 NLR 家族蛋白 3（NLRP3）在调节草酸钙一水合物（COM）诱导的人肾 HK-2 细胞损伤中的相互作用。用 COM（100μg/ml）处理 HK-2 细胞，建立肾损伤细胞模型。采用定量逆转录聚合酶链反应（qRT-PCR）和 Western blot 检测基因和蛋白表达。测定细胞增殖和凋亡率，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，利用 starBase 和 TargetScan 数据库预测与 SBF2-AS1 和 NLRP3 相互作用的潜在 miRNA。干扰 SBF2-AS1 可增加 COM 诱导后 HK-2 细胞的增殖和 SOD 水平。沉默 SBF2-AS1 还通过下调 NLRP3 蛋白表达，减少 COM 诱导的细胞死亡和炎症细胞因子的产生。相反，强制上调 NLRP3 可消除 SBF2-AS1 干扰的作用。值得注意的是，抗氧化剂可挽救 SBF2-AS1 干扰对 COM 诱导的氧化应激和 COM 诱导的细胞损伤的作用，表明氧化应激在 COM 诱导的损伤中起作用。miR-302e 作为一种介导 miRNA，连接 SBF2-AS1 和 NLRP3 的功能关联。沉默 SBF2-AS1 可促进 HK-2 细胞中 miR-302e 的水平，降低 miR-302e 对 NLRP3 的表达，从而保护 HK-2 细胞免受 COM 诱导的损伤。综上所述，这些发现表明下调 lncRNA SBF2-AS1 可能通过调节 miR-302e/NLRP3 通路来保护 HK-2 细胞免受 COM 诱导的损伤。
