摘要
目的:探讨人参三七川芎提取物对内皮细胞复制性衰老(humanumbilical vein endothelial cells,HUVECs)的影响。方法:体外培养的HUVEC-2C传代至第8代,造成内皮细胞复制性衰老模型。细胞分为4组,8代衰老模型组、维生素E(Vit E)组(50 mg.L-1)、中药高、低(50,20 mg.L-1)剂量组,采用衰老相关β-半乳糖苷酶(SA-β-gal)染色法观察细胞衰老改变,流式细胞仪分析细胞周期,激光共聚焦显微镜观察细胞内活性氧(ROS)含量,硝酸还原酶法检测培养液一氧化氮(NO)和抗超氧阴离子水平,Western-blot法检测各组细胞AngⅡ1型和2型受体(AT1R和AT2R),NAD(P)H氧化酶p47phox的蛋白表达。结果:8代衰老模型组β-gal染色阳性细胞数明显增多,细胞停滞在G0/G1期,激光共聚焦显微镜下ROS荧光强度明显增强,培养液中抗超氧阴离子、NO含量减少,p47phox,AT1R,AT2R蛋白表达上调。给予高、低剂量中药处理后,可明显改善细胞的衰老状态,β-gal染色阳性细胞数减少,G0/G1期细胞减少,G2/M期细胞增加,ROS荧光强度减弱,抗超氧阴离子、NO含量较8代衰老模型组增加,p47phox,AT1R,AT2R蛋白表达下调,差异均有统计学意义(均P<0.05)。结论:人参三七川芎提取物能够延缓内皮细胞复制性衰老,通过活性氧途径下调NADPH氧化酶p47phox的表达,最终使ROS产生减少;同时,中药提取物还能降低衰老内皮细胞AT1R,AT2R的表达,改善内皮功能,这可能是益气活血中药延缓内皮细胞衰老的主要原因之一。
Objective: To explore effect of extracts from Panax ginseng, P. notoginseng and Ligusticum chuanxiong on human umbilical endothelial cells (HUVECs) replicative senescence. Method: HUVECs were induced to aging by generation cultivating to the eighth cells in order to establish a model of endothelial ceils replicative senescence. The cultured HUVECs in vitro were divided into 4 groups, the eighth generation cell-senescence untreated group, Vitamin E group, herbal treated high dose and low dose groups. Changes of HUVECs aging were observed by method of SA-β-gal stained HUVECs and cells cycle were analyzed. Contents of ROS in cells,the levels of anti-superoxide (O^2- ) and nitric oxide (NO) in cell mediums were examined. Western blot were used to analyse protein expression of NADPH oxidase p47phox, angiotensin type 1 and 2 receptor (AT1 R, AT2R). Result: Compared with Vitamin E group, the positive cell numbers ofβ-gal stained HUVECs were enhanced, cell proliferation was depressed, and the fluorescence intensity of ROS was increased, at the same time, less NO and more O^2- in cells were produced in the eighth generation cell treated group: Protein expression of p47phox, AT1 R and AT2R in cells increased compared with Vit E group. Chinese herbs of high dose and low dose could improve condutions of HUVECs aging. Chinese herbs of high dose and low dose could reduce the positive cell numbers of β-gal stained HUVEC, increase cell proliferation and decrease fluorescence intensity of ROS in cells, at the same time, cells secreting more NO and less O^2-. Protein expression of p47phox, AT1R and AT2R in cells treated with Chinese herbs of high dose and low dose were decreased compared with Vit E group. Conclusion: The study indicated that extracts from P. ginseng, P. notoginseng and L. chuanxiong could delay endothelial cell replicative senescence. Herbal extracts downregulate the expression of NAD (P) H oxidase subunit-p47phox by means of ROS, hence decrease O^2- production and ultimately delay HUVECs in vitro senescence.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2009年第12期1544-1548,共5页
China Journal of Chinese Materia Medica
基金
国家重点基础研究发展计划(973)项目(2007CB507406)
关键词
人参三七川芎提取物
NAD(P)H氧化酶
细胞复制性衰老
ROS
extracts from Panax ginseng
Panax notoginseng and Ligusticum chuanxiong
NAD (P) H oxidase
cell replicative senescence
ROS