Department of Gastroenterology, Chongqing Wanzhou Shanghai Hospital, Chongqing, 404100, People's Republic of China.
Department of Gastroenterology, Chongqing Jiulongpo District People's Hospital, 7 Metallurgical Third Village, Shipingqiao, Jiulongpo, Chongqing, 400000, People's Republic of China.
Biochem Genet. 2022 Dec;60(6):2286-2298. doi: 10.1007/s10528-022-10219-2. Epub 2022 Mar 24.
Mounting evidence suggests that long non-coding RNAs (lncRNAs) and microRNAs exert a critical regulatory role in acute pancreatitis. The present study aimed to explore the role of lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) in acute pancreatitis (AP) that was induced by caerulein in rat pancreatic acinar cells (AR42J). The potential target sites of lncRNA NEAT1 and miR-365a-3p were predicted using starBase and were confirmed using dual-luciferase reporter assay. Reverse transcription-quantitative polymerase chain reaction was performed to assess lncRNA NEAT1 and miR-365a-3p expression levels in AP induced by caerulein. Cell Counting Kit-8 and flow cytometry assays were performed to assess AR42J cell viability. Western blotting was performed to evaluate the expression of apoptosis-related proteins. Interleukin (IL)-1β, IL-6, and tumor necrosis factor-α levels were detected by ELISA. The results of the dual-luciferase reporter assay confirmed that miR-365a-3p could bind to NEAT1. LncRNA NEAT1 was upregulated in AR42J cells treated with 10 nmol/l caerulein, and miR-365a-3p was expressed at low levels in an AP model. Overexpression of miR-365a-3p suppressed the apoptosis and inflammatory response of AR42J cells induced by caerulein. Importantly, inhibition of lncRNA NEAT1 decreased apoptosis and inflammation in caerulein-treated AR42J cells, while these effects were reverted upon co-transfection with a miR-365a-3p inhibitor. In conclusion, lncRNA NEAT1 was involved in AP progression by sponging miR-365a-3p and may thus be a novel target for treating patients with AP.
越来越多的证据表明，长链非编码 RNA（lncRNA）和 microRNA 在急性胰腺炎中发挥着关键的调节作用。本研究旨在探讨 lncRNA 核斑组装转录物 1（NEAT1）在 Caerulein 诱导的大鼠胰腺腺泡细胞（AR42J）急性胰腺炎（AP）中的作用。使用 starBase 预测 lncRNA NEAT1 和 miR-365a-3p 的潜在靶位点，并使用双荧光素酶报告基因检测进行验证。逆转录定量聚合酶链反应（RT-qPCR）检测 Caerulein 诱导的 AP 中 lncRNA NEAT1 和 miR-365a-3p 的表达水平。细胞计数试剂盒-8 和流式细胞术检测 AR42J 细胞活力。Western blot 检测凋亡相关蛋白表达。酶联免疫吸附试验（ELISA）检测白细胞介素（IL）-1β、IL-6 和肿瘤坏死因子-α水平。双荧光素酶报告基因检测的结果证实，miR-365a-3p 可与 NEAT1 结合。用 10 nmol/L Caerulein 处理的 AR42J 细胞中 lncRNA NEAT1 上调，AP 模型中 miR-365a-3p 表达水平较低。过表达 miR-365a-3p 可抑制 Caerulein 诱导的 AR42J 细胞凋亡和炎症反应。重要的是，抑制 lncRNA NEAT1 可降低 Caerulein 处理的 AR42J 细胞中的凋亡和炎症，而共转染 miR-365a-3p 抑制剂可逆转这些作用。综上所述，lncRNA NEAT1 通过海绵吸附 miR-365a-3p 参与 AP 进展，因此可能成为治疗 AP 患者的新靶点。
