| 杨静,雷燕,方素萍,崔巍,陈可冀.人参三七川芎提取物延缓血管紧张素Ⅱ诱导内皮细胞衰老机制的研究[J].,2009,(6):524-528 |
| 人参三七川芎提取物延缓血管紧张素Ⅱ诱导内皮细胞衰老机制的研究 |
| Study on Acting Mechanism of Extracts from Ginseng,Notoginseng and Chuanxiong for Delaying the Aging of Endothelial Cells Induced by AngiotensinⅡ |
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| DOI： |
| 中文关键词: 人参三七川芎提取物 血管紧张素Ⅱ NAD(P)H氧化酶 内皮细胞 衰老 细胞内活性氧 |
| 英文关键词:extracts from ginseng notoginseng and chuanxiong angiotensinⅡ NAD(P)H oxidase endothelial cell aging reactive oxygen species |
| 基金项目:国家重点基础研究发展计划(973计划)项目“衰老的机制与干预的基础研究”(No.2007CB507406) |
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| 中文摘要: |
| 目的探讨人参三七川芎提取物对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的内皮细胞(human umbilical vein endothelial cells,HUVECs)衰老的影响。方法体外培养的HUVECs加入终浓度为10-6mol/LAngⅡ诱导细胞衰老,造成内皮细胞衰老模型。细胞分为AngⅡ衰老模型组、空白对照组、人参三七川芎高剂量组、人参三七川芎低剂量组和缬沙坦组,采用SA-β-半乳糖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AngⅡ衰老模型组与空白对照组比较,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蛋白表达明显上调,AT1R表达较空白对照组上调。给予高、低剂量中药处理后改善了细胞的衰老状态,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AngⅡ衰老模型组增加,p47phox蛋白表达下调,AT1R表达较AngⅡ衰老模型组下调。结论人参三七川芎提取物能够延缓AngⅡ诱导的HUVECs衰老,可通过AT1R下调NAD(P)H氧化酶p47phox表达,进而使ROS产生减少,可能是益气活血中药延缓内皮细胞衰老的主要原因之一。 |
| 英文摘要: |
| Objective To explore the effects of extracts from ginseng,notoginseng and chuanxiong (Ext) on the angiotensin Ⅱ induced aging of human umbilical endothelial cells (HUVECs) in vitro. Methods Cultured HUVECs were divided into 5 groups,the blank control group (A),the model group (B),and the three intervening groups (C,D and E). Except those in Group A,all cells were induced into aging model cells by AngⅡ in a final concentration of 10-6 mol/L,and to the three intervening groups,corresponding treatments with low dose Ext (C),high dose Ext (D) and valsartan (E) were accessed. Changes of aging in HUVECs were observed by SA-β-gal staining;cell cycle was analyzed by flow cytometry;contents of reactive oxygen species (ROS) in cells was measured by laser confocal microscopy;levels of nitric oxide (NO) and anti-superoxide anion in culture medium were examined by nitrate reductase method;and the protein expression of NAD(P)H oxidase p47phox,as well as the angiotensin type 1 and 2 receptor (AT1R,AT2R) were detected by Western blot. Results Compared with Group A,in Group B,the positive β-gal stained HUVECs increased and stagnated at G0-G1 phase,with the fluorescence intensity of ROS evidently enhanced;in the culture medium content of NO and anti-superoxide anion were lowered,and protein expression of p47phox and AT1R up-regulated. While these aging figures were improved in Group C and D. After intervention with high or low dose of Ext,β-gal stained HUVECs decreased showing less cells in G0-G1 phase and more cells in G2-M phase,the fluorescence intensity of ROS reduced,contents of NO and anti-superoxide anion increased,and the protein expression of p47phox and AT1R down-regulated. Conclusion Ext could delay the aging of HUVECs induced by angiotensinⅡ,it is possibly by way of down-regulating the expression of NAD(P)H oxidase subunit-p47phox through AT1R,and further reducing the superoxide anion production. |
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