Senescent endothelial dysfunctions were mediated by S1P2 receptor in cultured human umbilical vein endothelial cells.
10.3969/j.issn.1672-7347.2012.12.010
- Author:
Shuhua CHEN
1
;
Hong XIANG
;
Guoping YANG
;
Hao DENG
;
Hong YUAN
;
Hongwei LU
Author Information
1. Department of Biochemistry, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Cells, Cultured;
Cellular Senescence;
genetics;
Human Umbilical Vein Endothelial Cells;
cytology;
physiology;
Humans;
RNA Interference;
RNA, Small Interfering;
genetics;
Receptors, Lysosphingolipid;
genetics;
metabolism;
Sphingosine-1-Phosphate Receptors;
Transfection;
Up-Regulation
- From:
Journal of Central South University(Medical Sciences)
2012;37(12):1239-1245
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the variation of senescent endothelial function by regulating the sphingosine-1-phosphate receptor type 2 (S1P2) expression in cultured human umbilical vein endothelial cells (HUVECs).
METHODS:The S1P2 receptor expression was regulated by transfecting the cDNA or shRNA of S1P2 in cultured HUVECs. The expression levels of S1P2 receptor in HUVECs were detected by RT-PCR and Western blot. EC chemotaxis was measured by the transwell migration assay. The wound healing assay was performed by a scratch wound model on EC monolayer. Matrigel morphogenesis assay was employed to assess the in vitro angiogenic responses.
RESULTS:After up-regulating the S1P2 expression in young ECs, the S1P-stimulated formation of a tubular-like network in Matrigel was dramatically diminished in transfected ECs (P<0.05). Quantification of the wound healing assay showed that transfected ECs grew much slower than young ECs (P<0.05). The chemotactic capability was significantly decreased in transfected ECs (P<0.05). Furthermore, the senescent-associated impairments were revoked by the downregulation of S1P2 receptor in senescent HUVECs.
CONCLUSION:The impaired functions (chemotactic, wound-healing and morphogenetic responses) in senescent HUVECs in vitro are mediated by S1P2 receptor.
