目的:探讨益气活血中药对血管内皮细胞衰老和p53基因表达的干预作用。方法:分离人脐静脉内皮细胞(HUVEC),传代培养至第4代,血管紧张素Ⅱ(AngⅡ,10-6mol/L,48h)诱导其衰老;分为空白对照组、AngⅡ组、中药提取物组和替米沙坦组;观察各组不同时间点β-半乳糖苷酶染色、CCK-8细胞计数、细胞周期分析、p53 m RNA及蛋白表达。结果:AngⅡ组较空白对照组β-半乳糖苷酶阳性染色细胞数增加(P<0.05);CCK-8检测活细胞数减少(P<0.05);细胞大量停滞于G0/G1期,而S期和G2/M期逐渐消失(P<0.05)。中药提取物组和替米沙坦组β-半乳糖苷酶阳性染色细胞数下降(P<0.05),CCK-8活细胞数增加(P<0.05),细胞周期阻滞改善(P<0.05)。中药早期作用更加明显(P<0.05)。AngⅡ呈时间依赖性上调p53 m RNA和蛋白表达;中药提取物和替米沙坦两组p53表达均明显下调(P<0.05);中药早期作用更加明显(P<0.05)。结论:益气活血中药能够有效地延缓血管内皮细胞衰老,其机制可能与抑制p53基因表达有关。
Objective:To observe the time-effect relation of extracts from ginseng,notoginseng and chuanxiong on angiotensin II(AngⅡ)-induced senescence of vascular endothelial cells and explore the feature of Chinese medicine against vascular diseases.Methods:Human umbilical vein endothelial cells(HUVECs)cultured in vitro were stimulated with 10^(-6) mol/L Ang Ⅱ to induce cell senescence,which were divided into 4groups,the blank control group,the Ang Ⅱ model group,the extracts group and the telmisartan group.The(J-gal was used to identify senescence of cells,the cell counting kit-8 method was applied to assess the cell viability,the cell function was examined with the level of endothelial nitric oxide synthase(eNOS) and the flow cytometry was used for analyzing the cell cycle changes.Results:Compared with the control cells,the cells positive for β-gal staining was significantly increased in the Ang Ⅱ model group,and showed cell cycle arrest at G_0/G_1 phase with decreased S and G_2/M phase ceil percentage,eNOS expression and cell viability(P<0.05).The extracts and telmisartan treatment of Ang II-induced cells resulted in decreased β-gal positive cells with a reduction in G_0/G_1phase cells and an increasing in S,G_2/M phase ceils and eNOS expression(P<0.05).At 24 h,the extracts were more effective than telmisartan(P<0.05);while telmisartan was more effective at 48 h(P<0.05).Conclusion:Extracts from ginseng,notoginseng and chuanxiong can delay Ang II-induced aging of HUVECs and may play an important role in early senescence.
