CAJ | 학술논문

目的用过氧化氢碳酰胺(carbamide peroxide,CP)作为诱导剂,诱导猪髂动脉内皮细胞(pig iliac endothelium cells,PIEC)衰老,建立体外内皮细胞衰老模型.方法 MTT法及DAH染色法确定CP对PIEC作用的最佳诱导浓度.细胞经过诱导后,形态学观察、MTT法、EdU掺入实验、SA-β-gal染色以及细胞迁移实验分析诱导细胞增殖活性、衰老比例以及迁移能力改变.PI单染流式细胞术分析诱导前后细胞周期的分布变化.结果 40 μmol/L为CP诱导内皮细胞的最佳浓度,诱导1h后,细胞变大变圆,呈典型的衰老表现,增殖活性和迁移能力下降,SA-β-gal染色显示衰老细胞比例明显增大,流式细胞分析显示大量细胞阻滞于G0/G1期.结论 CP能够体外诱导血管内皮细胞衰老,是一种安全高效的体外细胞衰老诱导剂.
목적 용과양화경탄선알(carbamide peroxide,CP)작위유도제,유도저가동맥내피세포(pig iliac endothelium cells,PIEC)쇠로,건입체외내피세포쇠로모형.방법 MTT법급DAH염색법학정CP대PIEC작용적최가유도농도.세포경과유도후,형태학관찰、MTT법、EdU참입실험、SA-β-gal염색이급세포천이실험분석유도세포증식활성、쇠로비례이급천이능력개변.PI단염류식세포술분석유도전후세포주기적분포변화.결과 40 μmol/L위CP유도내피세포적최가농도,유도1h후,세포변대변원,정전형적쇠로표현,증식활성화천이능력하강,SA-β-gal염색현시쇠로세포비례명현증대,류식세포분석현시대량세포조체우G0/G1기.결론 CP능구체외유도혈관내피세포쇠로,시일충안전고효적체외세포쇠로유도제.
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.