摘要
目的探讨血管紧张素Ⅱ(AngⅡ)对人脐静脉内皮细胞(HUVECs)凋亡的影响。方法体外培养HUVECs,取对数生长期HUVECs细胞接种96孔培养板中,常规培养细胞24 h后,弃掉旧培养基,加入新Ham′s F12k培养基,并分别加入AngⅡ,使其终浓度分别为0μmol.L-1、0.01μmol.L-1、0.1μmol.L-1、1μmol.L-1和10μmol.L-1,采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测不同浓度AngⅡ在不同时间点的细胞活性,琼脂糖凝胶电泳和流式细胞术检测细胞凋亡情况。结果MTT结果:与对照组相比,1μmol.L-1AngⅡ组作用24 h和10μmol.L-1AngⅡ组作用12 h、18 h和24 h的HUVECs的细胞活性均显著下降(P<0.05或P<0.01),并呈现时间依赖性;其余各组差异均无统计学意义(P>0.05)。琼脂糖凝胶电泳结果:10μmol.L-1AngⅡ组作用18 h的HUVECs呈现明显的"梯状"DNA断裂条带。流式细胞术结果:对照组和10μmol.L-1AngⅡ组作用18 h的HUVECs细胞凋亡率分别为(1.11±0.54)%和(19.87±3.18)%,2组比较差异有统计学意义(P<0.05)。结论10μmol.L-1AngⅡ可通过诱导细胞凋亡引起HUVECs损伤,从而可能在动脉粥样硬化发生发展中发挥作用。
Objective To explore the effects of angiotensin Ⅱ （Ang Ⅱ） on apoptosis of human umbilical vein endothelial cells （HUVECs）. Methods The HUVECs were cultured in vitro. The cells in exponential growth phase were inoculated to 96 hole training board.24 h after conventional culturing,the nutrient medium was given up,and new Ham＇s F12k nutrient medium were added in it and AngU was added in it too. The final concentration of Ang Ⅱ was 0 μmol·L^-1,0. 01μmol·L^-1 ,0. 1μmol·L^-1, 1 μmol·L^-1and 10μmol·L^-1 respectively. The viability of ceils at different time was estimated by MTT assay. While apoptosis of HUVECs were detected by agarose gel electrophoresis and flow cytometry. Results The results from MTT assay indicated that,compared to the control group,the viabilities of the HUVECs treated with 1μmol·L^-1 Ang Ⅱ for 24 h and 10 μmol·L^-1 Ang U for 12 h, 18 h and 24 h were decreased significantly （P 〈 0.05 or P 〈 0.01 ）, there was no difference in other groups. The results from the agarose gel electrophoresis displayed that ＂DNA ladder＂ were seen in HUVECs treated with 10 μmol·L^-1 Ang Ⅱ for 18 h; The results from flow cytometry exhibited that the apoptotic rates of the control group and HUVECs treated with 10 v Ang Ⅱ for 18 h were （1.11 ± 0.54）% and （19.87 ± 3.18 ）% respectively, there was statistical significance between two groups （P 〈 0.05 ）, Conclusion 10μmol·L^-1 Ang Ⅱ can injure HUVECs by inducing apoptosis, and it may play an important role in the development of atherosclerosis.
出处
《新乡医学院学报》
CAS
2009年第3期257-259,共3页
Journal of Xinxiang Medical University
关键词
血管紧张素Ⅱ
人脐静脉内皮细胞
凋亡
angiotensin Ⅱ
human umbilical vein endothelial cell
apoptosis
