title: MCPIP is induced by cholesterol and participated in cholesterol-caused DNA damage in HUVEC.
abstract: Hypercholesterolemia is an important risk factor for atherosclerosis and cholesterol treatment would cause multiple damages, including DNA damage, on endothelial cells. In this work, we have used human umbilical vein endothelial cell line (HUVEC) to explore the mechanism of cholesterol induced damage. We have found that cholesterol treatment on HUVEC could induce the expression of MCPIP1. When given 12.5 mg/L cholesterol on HUVEC, the expression of MCPIP1 starts to increase since 4 hr after treatment and at 24 hr after treatment it could reach to 10 fold of base line level. We hypothesis this induction of MCPIP1 may contribute to the damaging process and we have used siRNA of MCPIP1 in further research. This MCPIP1 siRNA (siMCPIP) could down regulate MCPIP1 by 73.4% and when using this siRNA on HUVECs, we could see the cholesterol induced DNA damage have been reduced. We have detected DNA damage by γH2AX foci formation in nuclear, γH2AX protein level and COMET assay. Compare to cholesterol alone group, siMCPIP group shows much less γH2AX foci formation in nuclear after cholesterol treatment, less γH2AX protein level in cell and also less tail moment detected in COMET assay. We have also seen that using siMCPIP1 could result in less reactive oxygen species (ROS) in cell after cholesterol treatment. We have also seen that using siMCPIP could reduce the protein level of Nox4 and p47(phox), two major regulators in ROS production. These results suggest that MCPIP1 may play an important role in cholesterol induced damage.
authors.affiliation: Department of Biochemistry and Molecular Biology, Zunyi Medical College Zunyi, Guizhou, China ; Department of Medicine, Renal Division, Guizhou Provincial People's Hospital, Guizhou Provincial Institute of Nephritic & Urinary disease Guiyang, Guizhou, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Da
authors.affiliation: Department of Biochemistry and Molecular Biology, Zunyi Medical College Zunyi, Guizhou, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhuo
authors.affiliation: Department of Biochemistry and Molecular Biology, Zunyi Medical College Zunyi, Guizhou, China.
authors.forename: Minzhang
authors.initials: M
authors.lastname: Qian
mesh_terms: D002478:Cells, Cultured; D002784:Cholesterol / Q000633:toxicity*; D020552:Comet Assay; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D006657:Histones / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D006801:Humans; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000378:metabolism; D018384:Oxidative Stress; D034622:RNA Interference; D017382:Reactive Oxygen Species / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D015854:Up-Regulation
chemical_list: C468783:H2AX protein, human; D006657:Histones; D017382:Reactive Oxygen Species; D014157:Transcription Factors; D002784:Cholesterol; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498910:NOX4 protein, human; C060505:neutrophil cytosolic factor 1; D012260:Ribonucleases; C510646:ZC3H12A protein, human
keywords: DNA damage; HUVEC; MCPIP1; ROS; cholesterol; siMCPIP