人参三七川芎提取物激活过氧化物酶体增殖物激活受体γ延缓血管内皮细胞衰老的研究

Study on extracts from ginseng, sanqi and sichuan lovage rhizome on delaying vascular endothelial cells senescence via activating peroxisome proliferator activated receptor γ signaling

目的:探讨血管内皮细胞衰老过程中过氧化物酶体增殖物激活受体γ(PPARγ)的表达变化和益气活血中药人参三七川芎提取物的干预作用。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(telmisartan)组;观察不同时间点β-半乳糖苷酶染色、NO含量、PPARγm 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),NO含量明显下降(P<0.05)。与AngⅡ组比较,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),NO含量增加(P<0.05)。AngⅡ呈时间依赖性上调PPARγm RNA表达,但蛋白表达呈现早升后降趋势;中药提取物和替米沙坦两组PPARγ表达均明显上调(P<0.05),中药早期作用更为显著(P<0.05)。结论:益气活血中药能够延缓血管内皮细胞衰老,其机制可能与诱导PPARγ表达有关。

Objective： To explore the expression change of peroxisome proliferator activated receptor γ （PPARγ） in vascular endothelial cells senescence and intervention effect of extracts from ginseng, sanqi and siehuan lovage rhizome. Methods： Human umbilical vein endothelial cells （HUVECs） were separated and cultured in vitro until to the fourth generation. HUVECs were induced by angiotensin Ⅱ （AngⅡ, 10^-6mol/L） and lasted for 48h, then the cells were divided into control group, Angll group, Chinese herb extracts group and telmisartan group. The β-galactosidase staining, NO content and mRNA and protein expression of PPARγ at different time points were observed. Results： Compared with control group, the positive cell flumbers of β-gal staining were significantly increased in Angll group （P〈0.05）, and the content of NO was decreased in AngⅡ group （P〈0.05）. Chinese herb extracts and telmisartan could reduce the positive cell numbers of β-gal staining and increase the content of NO （P〈0.05）. AngⅡ could up-regulate the mRNA expression of PPARy and with time dependence, but protein expression in cells in AngⅡ group showed a trend of rising early and falling later. The PPARγ expression in cells of Chinese herb extracts group and telmisartan group was increased significantly （P〈0.05）, and the effect of Chinese herb extracts in regulation of PPARγ expression was more prominent in early stage （P〈0.05）. Conclusion： Herbs of benefiting qi for activating blood circulation could delay vascular endothelial cells senescence, and mechanism might be related with induction effect on PPARγ expression.