Effects and mechanism of knocking down lncRNA H19 to inhibit lipid accumulation in human THP-1 cells-derived macrophages.
- Author:
Xuemei WANG
1
,
2
;
Yue CHE
3
;
Jieying WANG
1
,
2
;
Ke MEN
4
Author Information
1. Department of Public Health, Xi'an Medical College
2. Shaanxi Key Laboratory of Ischemic Cardiovascular Diseases & Institute of Basic and Translational Medicine, Xi'an Medical College, Xi'an 710021, China.
3. Department of Public Health, Xi'an Medical College, Xi'an 710021, China.
4. Department of Public Health, Xi'an Medical College, Xi'an 710021, China. *Corresponding author, E-mail: menke@foxmail.com.
- Publication Type:Journal Article
- MeSH:
Humans;
Adenosine Triphosphate;
Cholesterol;
NF-kappa B;
PPAR alpha;
RNA, Long Noncoding/genetics*;
RNA, Small Interfering/genetics*;
THP-1 Cells;
Macrophages/metabolism*;
Lipid Metabolism
- From:
Chinese Journal of Cellular and Molecular Immunology
2023;39(10):884-890
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of long noncoding RNA H19 on lipid accumulation of macrophages under high fat stress and its mechanism. Methods Human THP-1 cells-derived macrophages were incubated with ox-LDL, and the effects of H19 siRNA intervention on lipid accumulation was observed. The THP-1 cells were divided into control group (conventional culture), ox-LDL group, siRNA negative control (NC siRNA) combined with ox-LDL treatment group, and H19 siRNA combined with ox-LDL treatment group. Oil red O staining was used to determine the lipid accumulation in cells, and cholesterol concentration was analyzed by enzymatic method; ATP assay kit for detecting celluar ATP content; colorimetry was used to detect the levels of oxidative stress indicators and ELISA was used to detect the levels of monocyte chemoattractant protein-1 (MCP-1) in the cell supernatant. Western blot analysis was used to detect the protein expression of ATP binding cassette transporter A1 (ABCA1), peroxisome proliferator-activated receptor α (PPARα), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and nuclear factor κB p-p65 (NF-κB p-p65). Results Knockdown H19 significantly inhibited intracellular lipid accumulation, decreased total cholesterol (TC) and cholesterol ester (CE) content, and decreased CE/TC ratio. Knockdown H19 significantly alleviated cell damage including an increase in ATP content, a decrease in oxidative stress levels and a decrease in MCP-1 levels, which caused by high-fat stress. H19 siRNA upregulated expression of ABCA1, PPARα and PGC-1α in THP-1 derived macrophages, downregulated NF-κB signal pathway. Conclusion Knockdown H19 upregulates PGC-1α expression in THP-1 cells and downregulates NF-κB pathway, which promotes cholesterol reverse transport, reduces inflammatory reaction and inhibits lipid accumulation.
