Ge Xiaoxiao, Sun Tao, Zhang Yanmei, Li Yongqing, Gao Peng, Zhang Dantong, Zhang Bingyang, Wang Peijun, Ma Wanshan, Lu Sumei
Department of Laboratory Medicine, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, 250014, Shandong, People's Republic of China.
Blood Transfusion Department, Henan Provincial People's Hospital, People's Hospital of Zhengzhou University, Zhengzhou, 450003, Henan, People's Republic of China.
Nutr Metab (Lond). 2022 Jan 12;19(1):3. doi: 10.1186/s12986-021-00634-4.
To investigate the differential expression profile of lncRNAs in the nonalcoholic fatty liver disease (NAFLD) model induced by oleic acid (OA) and to further explore the role of LINC01260 (ENST00000255183) in NAFLD, providing theoretical support for the clinical value of lncRNAs in NAFLD.
OA (50 μg/mL) was used to induce steatosis in normal human LO2 hepatocytes for 48 h and was verified by Oil red O staining. Differential expression profiles of lncRNAs were obtained by eukaryotic circular sequencing (RNA/lncRNA/circRNA-seq) techniques. A gain-of-function (GOF) strategy for LINC01260 was adopted, Oil red O staining and semiquantitative analysis were combined to explore whether the GOF of LINC01260 affects LO2 cell steatosis. CeRNA-based bioinformatics analysis of lncRNAs was performed, and the enriched mRNAs were further verified. RXRB siRNAs were applied and verify its role in LINC01260 regulated OA-induced hepatocytes steatosis.
Lipid droplets of different sizes were observed in the cells of the OA group. Absorbance in the OA group was significantly increased after isopropanol decolorization (P < 0.05). Compared with those in the control group, there were 648 lncRNAs with differential expression greater than 1 time in the OA group, of which 351 were upregulated and 297 were downregulated. Fluorescence quantitative PCR showed that the expression of LINC01260 in the OA group was downregulated by 0.35 ± 0.07-fold (P < 0.05). The formation of lipid droplets in LO2 cells of the LINC01260 GOF group decreased significantly (P < 0.05). CeRNA analysis indicated that the mRNA levels of RXRB, RNPEPL1, CD82, MADD and KLC2 were changed to different degrees. Overexpression of LINC01260 significantly induced RXRB transcription (P < 0.05) and translation, and RXRB silence attenuated the lipids decrease induced by LINC01260 overexpression.
The OA-induced NAFLD cell model has a wide range of lncRNA differential expression profiles. LINC01260 participates in the regulation of the lipid droplet formation process of NAFLD, and its overexpression can significantly inhibit the steatosis process of LO2 cells. Mechanistically, LINC01260 may act as a ceRNA to regulate the expression of RXRB, thereby affecting the adipocytokine signaling pathway.
研究油酸（OA）诱导的非酒精性脂肪性肝病（NAFLD）模型中lncRNAs的差异表达谱，并进一步探讨LINC01260（ENST00000255183）在NAFLD中的作用，为lncRNAs在NAFLD中的临床价值提供理论支持。
用OA（50μg/mL）诱导正常人LO2肝细胞脂肪变性48 h，并用油红O染色进行验证。通过真核环状测序（RNA/lncRNA/circRNA-seq）技术获得lncRNAs的差异表达谱。采用LINC01260的功能获得（GOF）策略，结合油红O染色和半定量分析，探讨LINC01260的GOF是否影响LO2细胞脂肪变性。对lncRNAs进行基于ceRNA的生物信息学分析，并对富集的mRNA进行进一步验证。应用RXRB siRNAs并验证其在LINC01260调节OA诱导的肝细胞脂肪变性中的作用。
OA组细胞中观察到不同大小的脂滴。异丙醇脱色后，OA组吸光度显著增加（P<0.05）。与对照组相比，OA组差异表达倍数大于1倍的lncRNAs有648个，其中上调351个，下调297个。荧光定量PCR显示，OA组LINC01260的表达下调0.35±0.07倍（P<0.05）。LINC01260 GOF组LO2细胞中脂滴形成明显减少（P<0.05）。ceRNA分析表明，RXRB、RNPEPL1、CD82、MADD和KLC2的mRNA水平有不同程度的变化。LINC01260的过表达显著诱导RXRB转录（P<0.05）和翻译，RXRB沉默减弱了LINC01260过表达诱导的脂质减少。
OA诱导的NAFLD细胞模型具有广泛的lncRNA差异表达谱。LINC01260参与NAFLD脂滴形成过程的调控，其过表达可显著抑制LO2细胞的脂肪变性过程。机制上，LINC01260可能作为ceRNA调节RXRB的表达，从而影响脂肪细胞因子信号通路。
