Microarray analysis was performed to compare the expression profiles of lncRNAs in the liver of NASH, NAFLD and normal mice (5 mice for each group). Methionine-choline-deficient Medium (MCD) with Lipopolysaccharide (LPS) or palmitic acid (PA)were used to built NASH cell models. The role and mechanism of LncRNA-gm9795 in NASH were explored by knocking down or over-expressing its expression.
A total of 381 lncRNAs were found to be not only highly expressed in NAFLD, but also is going to go even higher in NASH. A novel LncRNA-gm9795 was significantly highly expressed in liver tissues of NASH animal models and NASH cell models. By staining with Nile red, we found that gm9795 did not affect the fat accumulation of NASH. However, gm9795 in NASH cell models significantly promoted the expression of TNF [Formula: see text], IL-6, IL-1[Formula: see text], the important inflammatory mediators in NASH. At the same time, we found that gm9795 upregulated the key molecules in endoplasmic reticulum stress (ERS), while NF-[Formula: see text]B/JNK pathways were also activated. When ERS activator Thapsigargin (TG) was introduced in cells with Ggm9757 si-RNA, NF-[Formula: see text]B and JNK pathways were activated. Conversely, ERS inhibitor Tauroursodeoxycholic acid (TUDCA) inhibited NF-kB and JNK pathways in cells with gm9795 overexpression plasmid.
LncRNA-gm9795 promotes inflammatory response in NASH through NF-kB and JNK pathways by ERS, which might provide theoretical basis for revealing the pathogenesis of NASH and discovering new therapeutic targets.
进行微阵列分析以比较NASH肝脏中lncRNAs的表达谱,NAFLD和正常小鼠(每组5只小鼠)。使用含有脂多糖(LPS)或棕榈酸(PA)的甲硫氨酸-胆碱缺乏培养基(MCD)来建立NASH细胞模型。通过敲低或过表达LncRNA-gm9795在NASH中的作用和机制。
发现总共381个lncRNAs不仅在NAFLD中高度表达,但在纳什也会更高。新型LncRNA-gm9795在NASH动物模型和NASH细胞模型的肝组织中显著高表达。用尼罗河红染色,我们发现gm9795不影响NASH的脂肪积累。然而,GM9795在NASH细胞模型中显著促进TNF的表达[公式:见正文],IL-6,IL-1[公式:见正文],NASH中重要的炎症介质。同时,我们发现GM9795上调了内质网应激(ERS)的关键分子,而NF-[配方:参见正文]B/JNK途径也被激活。当ERS激活剂Thapsigargin(TG)与Ggm9757si-RNA一起引入细胞时,NF-[配方:参见正文]B和JNK途径被激活。相反,ERS抑制剂牛磺熊去氧胆酸(TUDCA)抑制具有gm9795过表达质粒的细胞中的NF-kB和JNK途径。
LncRNA-gm9795通过ERS通过NF-kB和JNK途径促进NASH中的炎症反应,为揭示NASH的发病机制和发现新的治疗靶点提供理论依据。