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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 investigate the intervention of Yiqi Huoxue Chinese medicine on vascular endothelial cell senescence and p53 gene expression. Methods: The human umbilical vein endothelial cells (HUVECs) were isolated and subcultured to the 4th passage. Angiotensin Ⅱ (Ang Ⅱ, 10-6mol / L, 48h) Group and telmisartan group; β-galactosidase staining, CCK-8 cell counting, cell cycle analysis, p53 m RNA and protein expression were observed at different time points in each group. Results: The number of β-galactosidase positive cells in AngⅡ group increased (P <0.05), the number of viable cells in CCK-8 decreased (P <0.05) Period and G2 / M phase disappeared (P <0.05). The number of β-galactosidase positive cells in the extract group and the telmisartan group decreased (P <0.05), the number of viable cells in CCK-8 increased (P <0.05) and the cell cycle arrest improved (P <0.05) . The early role of Chinese medicine is more obvious (P <0.05). Ang Ⅱ induced p53 mRNA and protein expression in a time-dependent manner. The expression of p53 was significantly down-regulated in both extract and telmisartan groups (P <0.05). Conclusion: Yiqi Huoxue Chinese medicine can effectively delay the aging of vascular endothelial cells, and its mechanism may be related to the inhibition of p53 gene expression.