Abstract 573: Human Microrna-548p Decreases apoB Secretion and Lipid Synthesis

Objective: MicroRNAs (miRs) play important regulatory roles in lipid and lipoprotein metabolism. ApoB, as the only essential scaffolding protein in the assembly of very low density lipoproteins, is a target to treat hyperlipidemia and atherosclerosis. We aimed to find out miRs that reduce apoB expression. Approach: Bioinformatics analyses predicted that hsa-miR-548p can interact with apoB mRNA.MiR-548p mimic and control were transfected in human and mouse hepatoma cell lines to test its role in regulating apoB secretion and mRNA expression levels. Site-directed mutagenesis was used to identify the interacting site of miR-548p in human apoB 3′-untranslated region. Fatty acid oxidation and lipid syntheses were examined in miR-548p overexpressing cells to investigate its function in lipid metabolism. Results: Experimentally, we observed that miR-548p significantly reduces apoB secretion from human hepatoma cells in time and dose dependent manner. Mechanistic studies showed that miR-548p interacts with the 3′-untranslated region of human apoB mRNA to enhance posttranscriptional degradation. Bioinformatics algorithms suggested two potential binding sites of miR-548p on human apoB mRNA. Site-directed mutagenesis studies revealed that miR-548p targets site II involving both seed and supplementary sequences. MiR-548p had no effect on fatty acid oxidation but significantly decreased lipid synthesis in human hepatoma cells by reducing the expression of HMGCR and ACSL4 enzymes involved in cholesterol and fatty acid synthesis. In summary, miR-548p reduces lipoprotein production and lipid synthesis by reducing expression of different genes in human hepatoma cells. Conclusion: These studies suggest that miR-548p could be useful in treating atherosclerosis, hyperlipidemia and hepatosteatosis.

Download Full-text

Human MicroRNA-548p Decreases Hepatic Apolipoprotein B Secretion and Lipid Synthesis

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.117.309247 ◽

2017 ◽

Vol 37 (5) ◽

pp. 786-793 ◽

Cited By ~ 15

Author(s):

Liye Zhou ◽

M. Mahmood Hussain

Keyword(s):

Fatty Acid ◽

Apolipoprotein B ◽

Untranslated Region ◽

Lipid Synthesis ◽

Site Directed Mutagenesis ◽

Acid Oxidation ◽

Human Hepatoma ◽

Human Hepatoma Cells ◽

Apob Mrna ◽

Apob Secretion

Objective— MicroRNAs (miRs) play important regulatory roles in lipid metabolism. Apolipoprotein B (ApoB), as the only essential scaffolding protein in the assembly of very-low-density lipoproteins, is a target to treat hyperlipidemia and atherosclerosis. We aimed to find out miRs that reduce apoB expression. Approach and Results— Bioinformatic analyses predicted that hsa-miR-548p can interact with apoB mRNA. MiR-548p or control miR was transfected in human and mouse liver cells to test its role in regulating apoB secretion and mRNA expression levels. Site-directed mutagenesis was used to identify the interacting site of miR-548p in human apoB 3′-untranslated region. Fatty acid oxidation and lipid syntheses were examined in miR-548p overexpressing cells to investigate its function in lipid metabolism. We observed that miR-548p significantly reduces apoB secretion from human hepatoma cells and primary hepatocytes. Mechanistic studies showed that miR-548p interacts with the 3′-untranslated region of human apoB mRNA to enhance post-transcriptional degradation. Bioinformatic algorithms suggested 2 potential binding sites of miR-548p on human apoB mRNA. Site-directed mutagenesis studies revealed that miR-548p targets site I involving both seed and supplementary sequences. MiR-548p had no effect on fatty acid oxidation but significantly decreased lipid synthesis in human hepatoma cells by reducing HMGCR (3-hydroxy-3-methylglutaryl-coenzyme A reductase) and ACSL4 (Acyl-CoA synthetase long-chain family member 4) enzymes involved in cholesterol and fatty acid synthesis. In summary, miR-548p reduces lipoprotein production and lipid synthesis by reducing expression of different genes in human liver cells. Conclusions— These studies suggest that miR-548p regulates apoB secretion by targeting mRNA. It is likely that it could be useful in treating atherosclerosis, hyperlipidemia, and hepatosteatosis.

Download Full-text

Ethanol and C2 ceramide activate fatty acid oxidation in human hepatoma cells

Scientific Reports ◽

10.1038/s41598-018-31025-0 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 2

Author(s):

Jason M. Correnti ◽

Lauren Gottshall ◽

Annie Lin ◽

Bianca Williams ◽

Amanke Oranu ◽

...

Keyword(s):

Fatty Acid ◽

Fatty Acid Oxidation ◽

Hepatoma Cells ◽

Acid Oxidation ◽

Human Hepatoma ◽

Human Hepatoma Cells

Download Full-text

Microtubular integrity differentially modifies the saturated and unsaturated fatty acid metabolism in cultured hep G2 human hepatoma cells

Lipids ◽

10.1007/s11745-005-1462-5 ◽

2005 ◽

Vol 40 (10) ◽

pp. 999-1006

Author(s):

Carlos A. Marra ◽

María J. T. de Alaniz

Keyword(s):

Fatty Acid ◽

Fatty Acid Metabolism ◽

Unsaturated Fatty Acid ◽

Acid Metabolism ◽

Hepatoma Cells ◽

Human Hepatoma ◽

Hep G2 ◽

Human Hepatoma Cells

Download Full-text

Berberine ameliorates fatty acid-induced oxidative stress in human hepatoma cells

Scientific Reports ◽

10.1038/s41598-017-11860-3 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 18

Author(s):

Yixuan Sun ◽

Xinlu Yuan ◽

Feifei Zhang ◽

Yamei Han ◽

Xinxia Chang ◽

...

Keyword(s):

Oxidative Stress ◽

Fatty Acid ◽

Hepatoma Cells ◽

Human Hepatoma ◽

Human Hepatoma Cells

Download Full-text

Cheese can reduce indexes that estimate fatty acid desaturation. Results from the Oslo Health Study and from experiments with human hepatoma cells

Applied Physiology Nutrition and Metabolism ◽

10.1139/h11-123 ◽

2012 ◽

Vol 37 (1) ◽

pp. 31-39 ◽

Cited By ~ 9

Author(s):

Arne T. Høstmark ◽

Marianne S.H. Lunde

Keyword(s):

Fatty Acid ◽

Hepg2 Cells ◽

Causal Effect ◽

Fatty Acid Distribution ◽

Hepatoma Cells ◽

Fatty Acid Desaturation ◽

Health Study ◽

Human Hepatoma ◽

Human Hepatoma Cells ◽

Δ9 Desaturase

Previously, cheese intake was shown to be inversely related to serum triglycerides, raising the possibility that cheese might inhibit triglyceride synthesis, which is governed by fatty acid desaturases. Therefore, analyses were done to study whether cheese intake was associated with indexes that reflect fatty acid desaturation in 121 healthy ethnic Norwegians aged 40–45 years, a subsample from the Oslo Health Study (N = 18 777). Experiments with human hepatoma cells (HepG2) were done to clarify whether cheese might have a causal effect on desaturases. Fatty acid distribution in lipids of human sera and HepG2 cells was determined by gas chromatography. Δ9-Desaturase was estimated by the (16:1,n-7)/(16:0) and (18:1,n-9)/(18:0) ratios, abbreviated ds9_1 and ds9_2, and Δ5-desaturase (ds5) by the (20:4,n-6)/(18:2,n-6) ratio. Correlation, ANOVA, and multiple linear regression models were used to study associations. Oslo Health Study: Subjects with cheese intake >4–6 times per week had 33% lower ds9_1 and 16% lower ds5 than subjects with intake ≤ 4–6 times per week. The cheese intake vs. ds5 association prevailed when adjusting for sex, time since last meal, fatty fish, vegetables, fruit–berries, fruit juice, cod liver oil, coffee, alcohol, body mass index, physical activity, length of education, and smoking. HepG2 cells: An ethanol extract of Jarlsberg cheese lowered the desaturase indexes. Inhibition of ds9_1 increased with increasing amount cheese extract added. Thus, cheese may contain inhibitors of desaturases, thereby providing an explanation for the previously reported negative association between cheese intake and triglycerides.

Download Full-text