Ryanodine downregulates the expression of p-eNOS (Thr495) and improves the functions of rapamycin treated endothelial outgrowth cells
10.3760/cma.j.issn.0253-3758.2011.09.016
- VernacularTitle:理阿诺碱增强雷帕霉素诱导的内皮生长晕细胞功能并降低内皮型一氧化氮合酶( Thr495)的磷酸化水平
- Author:
Xiao-Lin SHAO
1
;
Huai-Qin ZHANG
;
Sheng YE
;
Yi-Nuo LIN
;
De-Ye YANG
;
Xue XIA
;
Xiao-Yan HUANG
;
Yan-Li ZHANG
Author Information
1. 温州医学院附属第一医院
- Keywords:
Coronary restenosis;
Sirolimus;
Ryanodine;
Endothelial outgrowth cell
- From:
Chinese Journal of Cardiology
2011;39(9):847-852
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effects of ryanodine on rapamycin treated endothelial outgrowth cells (EOCs). Methods The mononuclear cells were harvested from umbilical cord blood by Ficoll density gradient centrifugation, then induced into EOCs tnd expanded in vitro. The endothelial characteristics of EOCs were identified by immunostaining and fluorescent staining. The EOCs were pretreated with or without ryanodine ( 10 μmol/L) for 1 h, and then treated with or without rapamycin ( 10nmoL/L) for 24 h. Proliforation was evaluated by CCK8 and migration was measured by Transwell. The protein expression of EOCs was evaluated by immunobloting technique with total eNOS antibody and phosphoeNOS(Thr495) antibody. Results Compared with control group, the proliferation and migration capacities of EOCs were significantly reduced while the phosphorylation of eNOS ( Thr495 ) protein was significantly upregulated in rapamycin group( P ＜ 0. 05 ), expression of total eNOS was not affected by rapamycin ( P ＞0. 05). Compared with rapamycin group, the proliferation and migration capacities of EOCs were significantly increased and the phosphorylation of eNOS(Thr495) protein was significantly downregnlated in ryanodine + rapamycin group( P ＜0. 05). The proliferation and migration capacities, the phosphorylation of eNOS (Thr495) protein and the expression of total eNOS were not affected by ryanodine alone ( P ＞ 0. 05 ).ConclusionsRapamycin reduced proliferation and migration capacities while upregulated the phosphorylation of eNOS (Thr495) protein of EOCs and these effects could bepartly reversed by cotreatment with ryanodine.
