School of Environmental and Chemical Engineering, Shanghai University, Shanghai, 200444, China.
School of Environmental and Chemical Engineering, Shanghai University, Shanghai, 200444, China; Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, 361005, China.
Aquat Toxicol. 2023 Sep;262:106656. doi: 10.1016/j.aquatox.2023.106656. Epub 2023 Aug 12.
Increasing evidence suggests that long non-coding RNAs (lncRNAs) play pivotal roles in various biological processes. However, current studies on lncRNAs mostly focus on mammalian species, with little research on the functional roles of lncRNAs in teleost fish. Here, we identified a novel intergenic lncRNA (linc-93.2) in the head kidney primary macrophages of common carp (Cyprinus carpio) after exposure to a typical environmental endocrine disrupting chemical, bisphenol A (BPA). As a result, linc-93.2 was more than 3,619 bp in length and predominantly localized to the nucleus of primary macrophages other than cytoplasm, with the highest expression level in spleen followed by head kidney among different organs. Bioinformatic analysis predicted a cis-target gene, dennd1b, and 20 trans-target genes including hsp70, gna13 and rasgap, were potentially regulated by linc-93.2; NFκB and estrogen receptor (ERα) binding sites were located in the promoter region upstream of its transcription start site, which together suggested the involvement of linc-93.2 in immune and neurological functions in fish. Based on that, the expression level of linc-93.2 was determined in macrophages following acute lipopolysaccharide (LPS) and BPA treatments, both of which significantly induced linc-93.2 and IL-1β expression in cells. Moreover, a NF-κB inhibitor PDTC significantly reduced linc-93.2 expression in macrophages, but co-exposure of macrophages to PDTC with BPA or LPS could significantly rescue linc-93.2 expression, consistent with the observation on that LPS or BPA alone significantly induced both linc-93.2 and its target gene expression. Interestingly, linc-93.2 and its target gene expression was significantly suppressed by an ER antagonist ICI 182,780, however, the co-exposure of macrophages to ICI 182,780 with BPA failed to attenuate their declined expression. Overall, the current study demonstrated that linc-93.2, a novel immune-related lncRNA, may participate in the immune processes of common carp macrophages via the NF-κB and ER pathway. The results presented in this study enhance our understanding of the immunotoxin mechanisms of BPA in teleost fish.
越来越多的证据表明，长非编码 RNA（lncRNA）在各种生物过程中发挥着关键作用。然而，目前关于 lncRNA 的研究主要集中在哺乳动物物种上，对鱼类中 lncRNA 的功能作用研究甚少。在这里，我们在鲤鱼（Cyprinus carpio）头肾原代巨噬细胞中鉴定了一种新型的基因间 lncRNA（linc-93.2），该基因在暴露于典型的环境内分泌干扰化学物质双酚 A（BPA）后表达上调。结果表明，linc-93.2 的长度超过 3619bp，主要定位于原代巨噬细胞的核内而不是细胞质内，在不同器官中脾脏的表达水平最高，其次是头肾。生物信息学分析预测 cis 靶基因 dennd1b，以及 20 个 trans 靶基因包括 hsp70、gna13 和 rasgap，可能受 linc-93.2 调控；NFκB 和雌激素受体（ERα）结合位点位于其转录起始位点上游的启动子区域，这表明 linc-93.2 参与了鱼类的免疫和神经功能。基于这一点，我们在急性脂多糖（LPS）和 BPA 处理后确定了 linc-93.2 在巨噬细胞中的表达水平，这两种处理都显著诱导了细胞中 linc-93.2 和 IL-1β的表达。此外，NF-κB 抑制剂 PDTC 显著降低了巨噬细胞中 linc-93.2 的表达，但 PDTC 与 BPA 或 LPS 共同暴露于巨噬细胞中可以显著挽救 linc-93.2 的表达，这与 LPS 或 BPA 单独显著诱导 linc-93.2 及其靶基因表达的观察结果一致。有趣的是，linc-93.2 及其靶基因的表达被雌激素拮抗剂 ICI 182,780 显著抑制，但巨噬细胞中 ICI 182,780 与 BPA 的共同暴露并不能减轻其表达的下降。总的来说，本研究表明，新型免疫相关 lncRNA linc-93.2 可能通过 NF-κB 和 ER 途径参与鲤鱼巨噬细胞的免疫过程。本研究的结果增强了我们对 BPA 在鱼类中的免疫毒素机制的理解。
