The establishment and validation of an endothelial cell senescence model induced by carbamide peroxide
10.3760/cma.j.issn.0253-3758.2012.11.012
- VernacularTitle:过氧化氢碳酰胺诱导血管内皮细胞衰老模型的建立和验证
- Author:
Quan-Bo ZHANG
1
;
Bi-Hua WU
;
Yu-Feng QING
Author Information
1. 川北医学院附属医院老年科
- Keywords:
Endothelium,vascular;
Cell aging;
Carbamide peroxide
- From:
Chinese Journal of Cardiology
2012;40(11):952-957
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an in vitro pig iliac artery endothelial cells (PIECs) senescence model using carbamide peroxide (CP).Methods MTT assay and DAPI staining were used to define the optimal concentration of CP for inducing to the PIECs senescence model.Cellular morphology,MTT assay,EdU labeling,SA-β-gal staining and cell scratch test were performed to analyze the cell growth kinetic,proliferative activity,aging ratio and migratory activity difference post CP induction.PI signal staining flow cytometry was used to analyze the cell cycle distribution difference of cells before and after CP induction.Results The optimal CP concentration was 40 μmoL/L to induce PIECs senescence.After 1 h treatment with 40 μmol/L CP,the PIECs presented typical aging form with lager and more rounded shapes.Compared with control group,the proliferative activity and the migratory distance of CP group were significantly decreased; the SA-β-gal staining positive ratio was significantly increased; the data of mitotic cycle distribution with flow cytometry analysis showed that most cells were arrested at G1/G0 phase.Conclusion CP could efficiently induce pig iliac artery endothelial cell senescence in vitro.
