Clinical Laboratory Center, Beijing Friendship Hospital, Capital Medical University, China.
Beijing Friendship Hospital, Capital Medical University, Beijing, China.
Bioengineered. 2021 Dec;12(1):6738-6747. doi: 10.1080/21655979.2021.1960463.
Nonalcoholic fatty liver disease (NAFLD) is characterized by high morbidity. Although long noncoding RNAs (lncRNAs) are known to have a role in NAFLD pathogenesis, the identified lncRNA types are limited. In this study, NAFLD models were established in vitro and in vivo using free fatty acid-treated LO2 cells and high-fat diet-fed mice, respectively. Microarray data were downloaded from the Gene Expression Omnibus database, and was selected for further analysis. Cell viability and apoptosis were measured using Cell Counting Kit 8 and flow cytometry assays. RNA expression was detected using reverse transcription-quantitative polymerase chain reaction. Triglyceride (TG) content and lipid deposition were detected using enzyme-linked immunosorbent assay and Oil-Red O staining. Western blotting was used to visualize protein expression. Starbase and TargetScan were used to predict the target miRNA and gene, and the predictions were verified through RNA pull-down and luciferase reporter assays. expression levels were significantly suppressed in NAFLD models, whereas overexpression inhibited lipogenesis-related gene () expression and TG/lipid accumulation in vitro. Subsequently, was predicted and verified to target , and its expression was notably increased in the NAFLD cell model. Moreover, transfection of antagonized the effects of on lipid formation and accumulation. was confirmed to be the target gene of and was downregulated in the NAFLD cell model. Silencing reversed the effects of the inhibitor on lipid formation and accumulation. inhibited NAFLD progression via the / axis. These findings may provide a novel strategy against NAFLD.
非酒精性脂肪性肝病（NAFLD）的发病率较高。虽然长链非编码 RNA（lncRNA）已被证实与 NAFLD 的发病机制有关，但已鉴定的 lncRNA 类型有限。在本研究中，分别通过游离脂肪酸处理的 LO2 细胞和高脂饮食喂养的小鼠在体外和体内建立了 NAFLD 模型。从基因表达综合数据库（Gene Expression Omnibus database）下载微阵列数据，并选择进行进一步分析。使用细胞计数试剂盒 8 和流式细胞术检测细胞活力和细胞凋亡。使用逆转录定量聚合酶链反应检测 RNA 表达。使用酶联免疫吸附试验和油红 O 染色检测甘油三酯（TG）含量和脂质沉积。使用 Western blot 可视化蛋白表达。使用 Starbase 和 TargetScan 预测靶 miRNA 和基因，并通过 RNA 下拉和荧光素酶报告基因检测验证预测。在 NAFLD 模型中， 表达水平显著降低，而过表达 则抑制了体外脂生成相关基因（）的表达和 TG/脂质积累。随后，预测并验证 靶向 ，并且其在 NAFLD 细胞模型中的表达明显增加。此外，转染 拮抗了 对脂质形成和积累的影响。 被证实是 的靶基因，并且在 NAFLD 细胞模型中下调。沉默 逆转了 抑制剂对脂质形成和积累的影响。 通过 / 轴抑制 NAFLD 进展。这些发现可能为治疗 NAFLD 提供一种新策略。
