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 effect of extracts from Radix Ginseng, Radix Notoginseng and Rhizoma Chuanxiong （EXT） on delaying vascular smooth muscle cells （VSMCs） aging in aged rats. Methods： VSMCs were obtained by the modified tissue explants technique and were shown to be positive for smooth muscle α-actin （SM-α-actin） by immunohistochemistry staining. VSMCs obtained from the young rats were served as the young control group; VSMCs obtained from the old rats were treated with no drug （the old group）, with low dose extracts （20 mg/L, the EXT low-concentration group） and high dose extracts （40 mg/L, the EXT highconcentration group）, and with Probucal （106 mol/L, the Probucal group） as a positive control. All groups were cultured for 24 h in the medium with 10% serum for 24 h followed by another 24 h in the serum-free medium. At the end of the 48-h culture, the following analyses were performed including determination of senescenceassociated β-galactosidase （SA β-Gal） activity, flow cytometry analysis of cell cycle, real-time quantitative reverse transcription polymerase chain reaction （RT-PCR） analyses of p16, Cyclin D1, cyclin-dependent kinase 4 （CDK4） and retinoblastoma （Rb） mRNA expression, and Western blotting analyses of p16, cyclin D1, CDK4 and phosphoretinoblastoma （pRb） protein expressions. Results： （1） In comparison to the younger rats, VSMCs from aged rats had significantly more SA β-Gal positive cells （P〈0.01） and more cells in S phase （P〈0.05）. VSMCs from the all treated groups showed a significant decrease in both SA β-Gal positive cells （P〈0.05） and S phase （P〈0.05） compared to the old rats. （2） Compared with the young group, VSMCs in the old group had a significant decrease in p16 and Rb mRNA expression and a significant increase in Cyclin D1 and CDK4 mRNA expression. Compared with the old group, VSMCs in the treated groups had a significant increase in p16 and Rb mRNA expression and a significant decrease in
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 G0/G1 phase with decreased S and G2/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 G0/G1phase cells and an increasing in S,G2/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.
p53 is an important target for studying vascular aging.However,as people gradually learned more about the miR-34s and the relationship between miR-34s and p53,new research idea emerged.This paper tries to elaborate the feature of p53,microRNA and miR-34s in-depth,analyze the regulatory action of miR-34s on p53,and offer some new prevention and treatment prospects about vascular aging in Chinese medicine.
