枸杞多糖对血管紧张素Ⅱ诱导的血管内皮细胞衰老及P53和P16表达的影响

摘要

Objective To investigate the role of lydum bararum polysaccharides (LBP) on angiotensin II (Angll)-induced senescence of human umbilical vein endothelial cells (HUVECs) and expressions of P53 and P16 and explore the mechanism of LBP against aging. Methods HUVECs cultured in vitro were stimulated with lxl0-6mmol/L Angll to induce cell senescence, which was identified using β-gal staining. Flow cytometry was used for analyzing the cell cycle changes, and the cell viability was assessed using CCK-8 method. Western blotting was employed to detect the expression of P53 and P16 in the exposed cells. Results Compared with the control cells, the cells positive for β-gal staining was significantly increased in Angll group, and showed cell cycle arrest at G0/G1 phase with decreased S-phase cell percentage and cell viability. The expression levels of P53 and P16 were significantly increased in the cells with Angll exposure (P<0.05). LBP treatment of Angll-exposed cells resulted in decreased β-gal-positive cells with a reduction in G0/G1 phase cells and an increase in S phase cells. LBP treatment also increased the cell viability and significantly decreased the expression levels of P53 and P16 (P<0.05). Conclusion LBP can delay Angll-induced aging of HUVECs possibly by down-regulating the expression of P53 and P15.%目的 观察枸杞多糖对血管紧张素II(AngII)诱导人脐静脉血管内皮细胞(HUVECs)衰老及对细胞衰老相关基因P53和P16表达的影响,以探讨其抗衰老作用及机制.方法 体外培养HuVEcs加入终浓度10-6mol/L AngII诱导细胞衰老,建立内皮细胞衰老模型.实验分为空白对照组、AngII模型组和枸杞多糖组;β-半乳糖苷酶染色法鉴定内皮细胞衰老状态;流式细胞技术分析细胞周期变化;CCK-8法检测细胞存活率情况;Western blotting法检测P53、P16蛋白的表达.结果 AngII模型组与空白对照组比较,β-gal阳性染色率明显增多,细胞周期多停滞于G0/G1期,S期细胞逐渐减少,细胞存活率明显下降,P53、P16蛋白表达水平明显上调.给予枸杞多糖处理后改善了细胞衰老状态,β-gal阳性染色率减少,G0/G1期细胞减少,S期细胞逐渐增多,细胞存活率增多,P53、P16蛋白的表达较AngII组下调.结论 枸杞多糖能够抑制AngII 诱导HUVECs衰老,其作用机制可能是通过下调P53及P16的表达而实现的.