Effects of miR-590-5p on ox-LDL-induced endothelial cells apoptosis and LOX-1 expression.
10.3969/j.issn.1672-7347.2012.07.005
- Author:
Bing QIN
1
;
Bo XIAO
;
Ting JIANG
;
Huan YANG
Author Information
1. Department of Neurology, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
genetics;
Cells, Cultured;
Human Umbilical Vein Endothelial Cells;
cytology;
Humans;
Lipoproteins, LDL;
pharmacology;
MicroRNAs;
genetics;
metabolism;
RNA, Messenger;
genetics;
metabolism;
Scavenger Receptors, Class E;
genetics;
metabolism;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2012;37(7):675-681
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of miR-590-5p on the expression of lectin-like oxidized low density lipoprotein receptor 1 (LOX-1) in apoptotic human umbilical vein endothelial cells (HUVECs) induced by ox-LDL, and to explore the role of miR-590-5p in modulating HUVECs apoptosis.
METHODS:HUVECs were exposed to ox-LDL (50 μg/mL) for 0 to 48 h. Apoptosis was detected by Annexin V-FITC stain and was distinguished from necrosis by propidium iodide (PI) staining. The relative expression level of miR-590-5p in HUVECs was analyzed using real-time quantitative PCR (RT-qPCR). HUVECs were transfected with miR-590-5p mimics or miRNA mimics control followed by 50 μg/mL ox-LDL stimulation for 48 h. LOX-1 mRNA and protein were measured by RT-qPCR and Western blot, and apoptosis in HUVECs was analyzed by flow ctyometry after Annexin V-FITC/PI double stain.
RESULTS:Incubation of HUVECs with 50 μg/mL ox-LDL for 0 to 48 h resulted in a time-dependent induction of apoptotic cell death and down-regulation of miR-590-5p. Transfection of miR-590-5p mimics suppressed LOX-1 expression at both mRNA and protein levels, leading to a reduction of ox-LDL-induced apoptosis in HUVECs.
CONCLUSION:MiR-590-5p protects endothelial cells from ox-LDL induced apoptosis by inhibiting the expression of LOX-1.