[The establishment and validation of an endothelial cell senescence model induced by carbamide peroxide].
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 G(1)/G(0) phase.


CONCLUSIONS
CP could efficiently induce pig iliac artery endothelial cell senescence in vitro.
Department of Geriatric, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Quan-bo
QB
Zhang
Bi-hua
BH
Wu
Yu-feng
YF
Qing
D000818:Animals; D000077463:Carbamide Peroxide; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D008955:Models, Cardiovascular; D010545:Peroxides / Q000494:pharmacology*; D013552:Swine; D014508:Urea / Q000031:analogs & derivatives* / Q000494:pharmacology
D010545:Peroxides; D000077463:Carbamide Peroxide; D014508:Urea