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Culture of Macrophage Colony-stimulating Factor Differentiated Human Monocyte-derived Macrophages
Published on: June 30, 2016
Mipu1 overexpression protects macrophages from oxLDL-induced foam cell formation and cell apoptosis
Shun-Lin Qu1, Wen-Jing Fan, Chi Zhang
11 Key Lab for Arteriosclerology of Hunan Province, Post-Doctoral Mobile Stations for Basic Medicine, Institute of Cardiovascular Disease, University of South China , Hengyang City, Hunan Province, People's Republic of China .
Abstract:
Mipu1 (myocardial ischemic preconditioning upregulated protein 1) is a novel N-terminal Kruppel-associated box (KRAB)/C2H2 zinc finger superfamily protein, that displays a powerful effect in protecting H9c2 cells from oxidative stress-induced cell apoptosis. The present study aims to investigate the effect of Mipu1 overexpression on oxidized low-density lipoprotein (oxLDL)-induced foam cell formation, cell apoptosis, and its possible mechanisms. New Zealand healthy rabbits were used to establish atherosclerosis model, and serum levels of triglycerides, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were detected by an automatic biochemical analyzer. Sudan IV staining was used to detect atherosclerotic lesions. The RAW264.7 macrophage cell line was selected as the experimental material. Oil red O staining, high-performance liquid chromatography, and Dil-labeled lipoprotein were used to detect cholesterol accumulation qualitatively and quantitatively, respectively. Flow cytometry was used to determine cell apoptosis. Real-time quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression of the main proteins that are associated with the transport of cholesterol, such as ABCA1, ABCG1, SR-BI, and CD36. Western blot analysis was used to detect the protein expression of Mipu1. There were atherosclerotic lesions in the high-fat diet group with Sudan IV staining. High-fat diet decreased Mipu1 expression and increased CD36 expression significantly at the 10th week compared with standard-diet rabbits. Mipu1 overexpression decreased oxLDL-induced cholesterol accumulation, oxLDL uptake, cell apoptosis, and cleaved caspase-3. Mipu1 overexpression inhibited the oxLDL-induced CD36 mRNA and protein expression, but it did not significantly inhibit the mRNA expression of ABCA1, ABCG1, and SR-BI. Mipu1 overexpression inhibits oxLDL-induced foam cell formation and cell apoptosis. Mipu1 overexpression reduces the lipid intake of macrophages and might be associated with the downregulation of CD36 expression in the presence of oxLDL.
Insights
Myeloid cell leukemia 1 (Mipu1) protein overexpression protects against atherosclerosis by reducing cholesterol accumulation and apoptosis. Mipu1 inhibits oxidized low-density lipoprotein (oxLDL) uptake and foam cell formation, potentially by downregulating CD36 expression.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Cellular Biology
Background:
- Oxidized low-density lipoprotein (oxLDL) plays a critical role in the development of atherosclerosis.
- Foam cell formation and macrophage apoptosis are key pathological processes in atherosclerosis.
- Mipu1 (myocardial ischemic preconditioning upregulated protein 1) is a novel protein with protective effects against oxidative stress.
Purpose of the Study:
- To investigate the effect of Mipu1 overexpression on oxLDL-induced foam cell formation and apoptosis.
- To elucidate the underlying mechanisms of Mipu1's action in atherosclerosis.
Main Methods:
- Atherosclerosis model in New Zealand rabbits and RAW264.7 macrophages.
- Biochemical analysis, Sudan IV, Oil red O, and Dil-labeled lipoprotein staining for lipid accumulation and lesions.
- Flow cytometry for cell apoptosis and Western blot/qPCR for protein and mRNA expression (Mipu1, CD36, ABCA1, ABCG1, SR-BI).
Main Results:
- High-fat diet induced atherosclerotic lesions and altered Mipu1 and CD36 expression in rabbits.
- Mipu1 overexpression reduced oxLDL-induced cholesterol accumulation, oxLDL uptake, and apoptosis in macrophages.
- Mipu1 overexpression downregulated CD36 mRNA and protein expression, but not ABCA1, ABCG1, or SR-BI.
Conclusions:
- Mipu1 overexpression inhibits oxLDL-induced foam cell formation and macrophage apoptosis.
- Mipu1 reduces lipid uptake in macrophages, likely through CD36 downregulation.
- Mipu1 shows therapeutic potential in mitigating atherosclerosis progression.
