Cholesterol Metabolism Regulation by Native and Oxidized Low-Density Lipoprotein in a Liver Cell Line
Abstract
AIMS: Metabolic dysfunction-associated steatohepatitis (MASLD) is a highly prevalent chronic liver disease. Oxidized low-density lipoprotein (oxLDL) is implicated in MASLD progression; however, its effects on the hepatic cholesterol metabolism remain unclear. Therefore, we explored the effects of native LDL (nLDL) and oxLDL on the cholesterol metabolism and lipid composition in the human liver-derived C3A cell line. METHODS: Orbitrap liquid chromatography-tandem mass spectrometry was used to measure free cholesterol (FC), cholesteryl ester (CE), and CE hydroperoxide (CEOOH) levels in LDL particles and LDL-treated C3A cells. The expression of cholesterol metabolic genes was examined using real-time PCR, and proprotein convertase subtilisin kexin type 9 (PCSK9) protein expression was quantified using Western blotting and an enzyme-linked immunosorbent assay. RESULTS: Both LDL types, particularly nLDL, increased intracellular CE levels. nLDL preferentially increased CE species containing unsaturated fatty acids, whereas oxLDL promoted the accumulation of saturated fatty acid (SFAs)-containing CEs and CEOOH, while FC levels remained unchanged. Both LDL types downregulated SREBP2 and downstream cholesterol synthesis genes and reduced LDLR expression. The PCSK9 protein levels also decreased. SOAT1 expression was reduced, indicating that CE accumulation was primarily driven by exogenous LDL-derived lipids rather than enhanced esterification. In addition, the genes involved in cholesterol efflux (ABCA1), xenobiotic metabolism, and scavenger receptor-related pathways were downregulated in C3A cells. CONCLUSION: Both nLDL and oxLDL particles drive CE accumulation through impaired cholesterol release rather than increased cholesterol synthesis. Furthermore, LDL oxidation increases CEOOH and saturated CEs, thus potentially enhancing hepatocellular lipotoxicity.
Related
- Upregulation of miR-200b-3p serves as a diagnostic biomarker for coronary atherosclerosis and regulates the oxidized low-density lipoprotein cholesterol-induced dysfunction of vascular smooth muscle cells
- Association between the fasting blood glucose to high-density lipoprotein cholesterol ratio and non-alcoholic fatty liver disease in Chinese adults: a secondary analysis
- Small dense low-density lipoprotein cholesterol and risk of cardiovascular events in general adults: a systematic review and meta-analysis of prospective studies.
- Association of the triglyceride-to-high-density lipoprotein cholesterol ratio with kidney stone status: cross-sectional and machine-learning analyses
- Joint association of the cholesterol–high-density lipoprotein–glucose index and waist-to-height ratio with incident cardiovascular disease in a high-risk Chinese cohort
BibTeX
@article{kim2026cholesterol,
title = {{Cholesterol Metabolism Regulation by Native and Oxidized Low-Density Lipoprotein in a Liver Cell Line}},
author = {Nakyoung Kim and Toshihiro Sakurai and Himeno Sakuma and Akiko Sakurai and Nanao Murakami and Iku Sazaki and Shu-Ping Hui},
journal = {Journal of Atherosclerosis and Thrombosis},
year = {2026},
month = sep,
doi = {10.5551/jat.66254},
url = {https://www.jstage.jst.go.jp/article/jat/advpub/0/advpub_66254/_article},
note = {CC BY-NC-SA},
}
RIS
TY - JOUR
TI - Cholesterol Metabolism Regulation by Native and Oxidized Low-Density Lipoprotein in a Liver Cell Line
AU - Nakyoung Kim
AU - Toshihiro Sakurai
AU - Himeno Sakuma
AU - Akiko Sakurai
AU - Nanao Murakami
AU - Iku Sazaki
AU - Shu-Ping Hui
PY - 2026
DA - 2026/09/08
JO - Journal of Atherosclerosis and Thrombosis
DO - 10.5551/jat.66254
UR - https://www.jstage.jst.go.jp/article/jat/advpub/0/advpub_66254/_article
AB - AIMS: Metabolic dysfunction-associated steatohepatitis (MASLD) is a highly prevalent chronic liver disease. Oxidized low-density lipoprotein (oxLDL) is implicated in MASLD progression; however, its effects on the hepatic cholesterol metabolism remain unclear. Therefore, we explored the effects of native LDL (nLDL) and oxLDL on the cholesterol metabolism and lipid composition in the human liver-derived C3A cell line. METHODS: Orbitrap liquid chromatography-tandem mass spectrometry was used to measure free cholesterol (FC), cholesteryl ester (CE), and CE hydroperoxide (CEOOH) levels in LDL particles and LDL-treated C3A cells. The expression of cholesterol metabolic genes was examined using real-time PCR, and proprotein convertase subtilisin kexin type 9 (PCSK9) protein expression was quantified using Western blotting and an enzyme-linked immunosorbent assay. RESULTS: Both LDL types, particularly nLDL, increased intracellular CE levels. nLDL preferentially increased CE species containing unsaturated fatty acids, whereas oxLDL promoted the accumulation of saturated fatty acid (SFAs)-containing CEs and CEOOH, while FC levels remained unchanged. Both LDL types downregulated SREBP2 and downstream cholesterol synthesis genes and reduced LDLR expression. The PCSK9 protein levels also decreased. SOAT1 expression was reduced, indicating that CE accumulation was primarily driven by exogenous LDL-derived lipids rather than enhanced esterification. In addition, the genes involved in cholesterol efflux (ABCA1), xenobiotic metabolism, and scavenger receptor-related pathways were downregulated in C3A cells. CONCLUSION: Both nLDL and oxLDL particles drive CE accumulation through impaired cholesterol release rather than increased cholesterol synthesis. Furthermore, LDL oxidation increases CEOOH and saturated CEs, thus potentially enhancing hepatocellular lipotoxicity.
SN - 1880-3873
SN - 1340-3478
ER -