长链非编码RNA Gm12664-001通过调节miR-295-5p和CAV1的表达改善非酒精性脂肪肝。
LncRNA Gm12664-001 ameliorates nonalcoholic fatty liver through modulating miR-295-5p and CAV1 expression.
作者信息
Zhang Qiao, Wang Jiemei, Li Hongyin, Zhang Yuan, Chu Xia, Yang Jianjun, Li Ying
机构信息
1Department of Nutrition and Food Hygiene, Public Health College, Harbin Medical University, Harbin, 150086 China.
2Department of Public Health College, Kunming Medical University, Kunming, 650550 China.
出版信息
Nutr Metab (Lond). 2020 Feb 4;17:13. doi: 10.1186/s12986-020-0430-z. eCollection 2020.
BACKGROUND
Our study aims to investigate the mechanisms of lncRNA Gm12664-001 improved hepatic lipid accumulation-initiated NAFLD via regulating miR-295-5p and CAV1 in AML12 cells.
METHODS
The animals were divided into normal control (NC) group and high fat diet (HFD) group (20 mice per group) for 8w. The steatotic liver was measured by hematoxylin eosin (HE) staining and kits. We performed systematical analyses on hepatic expression profiles of long noncoding RNAs (lncRNAs) and microRNAs in a high-fat diet (HFD)-induced steatotic animal model. The expression profile of targets was confirmed by bioinformatics analysis, luciferase assay, RT-PCR and western blot in AML12 cells.
RESULTS
HFD treatment markedly observed hepatic fatty degeneration with primarily fat vacuoles, and increased TG level compared with control. According to microarray data, we found that transfection of Gm12664-001 siRNA (siRNA-118,306) obviously enhanced TG accumulation and repressed CAV1 in AML12 cells. Furthermore, the TG accumulation markedly increased by siRNA-mediated knockdown of CAV1 in AML12 cells. By bioinformatics prediction, AML12 cells were transfected of siRNA-118,306 obviously upregulated miR-295-5p. Transfection of miR-295-5p mimics significantly increased TG accumulation and obviously suppressed the target CAV1.
CONCLUSIONS
The results revealed that lncRNA Gm12664-001 attenuated hepatic lipid accumulation through negatively regulating miR-295-5p and enhancing CAV1 expression in AML12 cells.
背景
我们的研究旨在探讨长链非编码RNA（lncRNA）Gm12664 - 001通过调控AML12细胞中的miR - 295 - 5p和小窝蛋白1（CAV1）改善肝脂质蓄积引发的非酒精性脂肪性肝病（NAFLD）的机制。
方法
将动物分为正常对照组（NC）和高脂饮食（HFD）组（每组20只小鼠），持续8周。通过苏木精 - 伊红（HE）染色和试剂盒检测脂肪肝。我们对高脂饮食（HFD）诱导的脂肪变性动物模型中的肝脏长链非编码RNA（lncRNAs）和微小RNA的表达谱进行了系统分析。通过生物信息学分析、荧光素酶报告基因检测、逆转录 - 聚合酶链反应（RT - PCR）和蛋白质免疫印迹法（western blot）在AML12细胞中确认靶标的表达谱。
结果
与对照组相比，高脂饮食处理明显观察到肝脏脂肪变性，主要为脂肪空泡，并增加了甘油三酯（TG）水平。根据微阵列数据，我们发现转染Gm12664 - 001小干扰RNA（siRNA - 118,306）明显增强了AML12细胞中的TG蓄积并抑制了CAV1。此外，在AML12细胞中，通过小干扰RNA介导的CAV1敲低显著增加了TG蓄积。通过生物信息学预测，转染siRNA - 118,306的AML12细胞明显上调了miR - 295 - 5p。转染miR - 295 - 5p模拟物显著增加了TG蓄积并明显抑制了靶标CAV1。
结论
结果表明，lncRNA Gm12664 - 001通过在AML12细胞中负调控miR - 295 - 5p并增强CAV1表达来减轻肝脏脂质蓄积。
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