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Effects and mechanism of knocking down lncRNA H19 to inhibit lipid accumulation in human THP-1 cells-derived macrophages / 细胞与分子免疫学杂志
Chinese Journal of Cellular and Molecular Immunology ; (12): 884-890, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1009444
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.
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Trifosfato de Adenosina / Colesterol / NF-kappa B / RNA Interferente Pequeno / PPAR alfa / Metabolismo dos Lipídeos / RNA Longo não Codificante / Células THP-1 / Macrófagos Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Cellular and Molecular Immunology Ano de publicação: 2023 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Trifosfato de Adenosina / Colesterol / NF-kappa B / RNA Interferente Pequeno / PPAR alfa / Metabolismo dos Lipídeos / RNA Longo não Codificante / Células THP-1 / Macrófagos Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Cellular and Molecular Immunology Ano de publicação: 2023 Tipo de documento: Artigo