| 摘要: |
| 目的通过体外细胞实验研究血管紧张素Ⅱ(Ang Ⅱ)对人脐静脉内皮细胞(HUVECs)分泌NO和ET1的影响,探讨其可能的机制。方法体外培养HUVECs,对其进行不同浓度Ang Ⅱ刺激(1×10-9、1×10-8、1×10-7、1×10-6mol/L刺激组),及相同浓度不同作用时间Ang Ⅱ刺激(1×10-7mol/L刺激2、6、12、18、24h),并应用不同的受体阻断剂(AT1R阻断剂替米沙坦、AT2R阻断剂PD123319)及信号通路阻断剂(Erk1/2阻断剂PD98059、P38 MAPK阻断剂SB203580、PKC阻断剂Staurosporine)观察Ang Ⅱ的作用通路,用ELISA法检测细胞分泌的内皮素1(ET1),用Griess法检测一氧化氮(NO),用RT-PCR方法检测eNOS/ET1 mRNA水平。结果 Ang Ⅱ对血管内皮细胞的刺激作用有浓度效应,Ang Ⅱ越高,eNOS mRNA表达和NO水平越低,而ET1 mRNA表达与蛋白水平越高；Ang Ⅱ对内皮细胞功能的影响有时间效应,Ang Ⅱ作用时间越长,NO/ET1水平越低；替米沙坦、PD98059和SB203580能阻断Ang Ⅱ的效应；而PD123319和Staurosporine不能阻断Ang Ⅱ的效应。结论 Ang Ⅱ可损伤内皮细胞的分泌功能,表现为NO降低而ET1升高,其通过AT1R发挥,用并可能通过Erk1/2及MAPK信号通路途径起作用。 |
| 关键词: 血管紧张素Ⅱ 内皮细胞功能 信号通路 |
| DOI：10.16118/j.1008-0392.2015.01.006 |
| 投稿时间：2014-02-17 |
| 基金项目: |
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| Effects of angiotensin Ⅱ on secretion of NO and ET1 in cultured human umbilical vein endothelial cells |
| QI Yuan,YAO Yi-an |
| (Dept. of Cardiology, East Hospital, Tongji University, Shanghai 200120, China) |
| Abstract: |
| Objective To investigate the effects of angiotensin Ⅱ(Ang Ⅱ) on secretion of nitric oxide(NO) and endothelin 1(ET1) in cultured human umbilical vein endothelial cells(HUVECs) and the underlying mechanisms. MethodsHUVECs were treated with Ang Ⅱ in different concentrations(1×10-9,1×10-8,1×10-7,1×10-6mol/L) or same concentration for different lengths of time(1×10-7mol/L for 2,6,12,8 and 24h), with or without Ang Ⅱ type 1 receptor(AT1R) blocker Telmisartan, Ang Ⅱ type 2 receptor(AT2R) blocker PD123319, or different kinase inhibitors PD98059(inhibitor of ERK1/2 MAP kinase), SB203580(inhibitor of ERK1/2 MAP kinase) and Staurosporine(phosphatidylinositol 3-kinase). Endothelin1(ET1) and NO in the medium were measured by ELISA and Griess reagent, respectively; and eNOS/ET1 mRNA expression was measured by RT-PCR. ResultsAng Ⅱ up-regulated ET-1 protein level and down-regulated NO concentration, and decrease the NO/ET1 ratio in HUVECs in a dose-and time-dependent manner within 24h. Telmisartan, PD98059 and SB203580 blocked this effect, but PD123319 and Staurosporine had no blockage effect. ConclusionAng Ⅱ can decrease the level of NO and increase the level of ET1, this effect were partly through AT1R and MAPK and Erk1/2 signal cell pathway. |
| Key words: angiotensin Ⅱ endothelial cell function cell signal pathway |
