College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China.
College of Pharmacy, Dalian Medical University, Western 9 Lvshunnan Road, Dalian 116044, China.
Int Immunopharmacol. 2024 Jan 25;127:111444. doi: 10.1016/j.intimp.2023.111444. Epub 2023 Dec 28.
Long non-coding RNAs (lncRNAs) have been demonstrated to play vital roles in the pathogenesis of nonalcoholic fatty liver disease (NAFLD). However, their biological roles and function mechanisms in NAFLD remain largely unknown. In this study, we found that Gm28382 may be a potential pathogenic lncRNA of NAFLD and highly expressed in NAFLD through RNA-seq. Overexpression of Gm28382 significantly enhanced the lipid accumulation in AML12 cells, whereas Gm28382 silencing reduced lipogenesis both in palmitic acid (PA)-induced AML12 cells and high fat diet (HFD)-induced mice. Then, bioinformatics were employed to speculate the potential interacting genes of Gm28382, and found that Gm28382 may regulate ChREBP expression through binding with miR-326-3p. Fluorescence in situ hybridization (FISH), dual luciferase reporter assay, immunofluorescence RNA pull-down and RNA immunoprecipitation (RIP) assays were used to validate the binding and targeting relationship of these genes, and we confirmed that Gm28382 competitively binds to miR-326-3p to increase ChREBP expression as a ceRNA. Mechanistically, overexpression of Gm28382 upregulated the ChREBP-mediated lipid synthesis signaling pathway, but the function was sabotaged by miR-326-3p deletion or ChREBP overexpression. Furthermore, in PA-challenged AML12 cells or HFD-induced mice, silencing of Gm28382 reversed the aberrant ChREBP signaling pathway and lipid accumulation, whereas ChREBP overexpression or liver-specific silencing of miR-326-3p blocked this function of Gm28382. Collectively, these findings reveal a critical role of Gm28382 in the promotion of lipogenesis in NAFLD by regulating the ChREBP signaling pathway through interaction with miR-326-3p, which could serve as a potential therapeutic target for NAFLD treatment.
长链非编码 RNA（lncRNA）已被证明在非酒精性脂肪性肝病（NAFLD）的发病机制中发挥重要作用。然而，它们在 NAFLD 中的生物学作用和功能机制在很大程度上仍然未知。在这项研究中，我们通过 RNA-seq 发现 Gm28382 可能是 NAFLD 的一种潜在致病 lncRNA，并且在 NAFLD 中高度表达。Gm28382 的过表达显著增强了 AML12 细胞中的脂质积累，而 Gm28382 的沉默减少了棕榈酸（PA）诱导的 AML12 细胞和高脂肪饮食（HFD）诱导的小鼠中的脂肪生成。然后，我们运用生物信息学方法推测 Gm28382 的潜在相互作用基因，并发现 Gm28382 可能通过与 miR-326-3p 结合来调节 ChREBP 的表达。荧光原位杂交（FISH）、双荧光素酶报告基因检测、免疫荧光 RNA 下拉和 RNA 免疫沉淀（RIP）实验用于验证这些基因的结合和靶向关系，我们证实 Gm28382 竞争性地与 miR-326-3p 结合，以作为 ceRNA 增加 ChREBP 的表达。在机制上，Gm28382 的过表达上调了 ChREBP 介导的脂质合成信号通路，但 miR-326-3p 的缺失或 ChREBP 的过表达破坏了这一功能。此外，在 PA 挑战的 AML12 细胞或 HFD 诱导的小鼠中，Gm28382 的沉默逆转了异常的 ChREBP 信号通路和脂质积累，而 ChREBP 的过表达或 miR-326-3p 的肝特异性沉默阻断了 Gm28382 的这一功能。总之，这些发现揭示了 Gm28382 通过与 miR-326-3p 相互作用调节 ChREBP 信号通路在促进 NAFLD 中脂质生成中的关键作用，这可能成为 NAFLD 治疗的潜在治疗靶点。
