Aβ_(1-42)诱导HUVEC细胞凋亡最适浓度与时间选择  

Screen of the optimal concentration and incubution time of Aβ_(1-42) in inducing apoptosis of HUVEC cell

作　　者：蔡鹏飞[1] 刘明华[2] 冯林林[3] 刘俊红[3] 尤朝菲 章卓[2] 

机构地区：[1]泸州医学院科技处,四川泸州646000 [2]泸州医学院药学院药理教研室,四川泸州646000 [3]泸州医学院

出　　处：《泸州医学院学报》2014年第2期169-171,共3页Journal of Luzhou Medical College

基　　金：四川省教育厅课题(102c099);泸州市科技局课题(2012s36)

摘　　要：目的:探讨β样淀粉肽1-42(Aβ1-42)诱导人脐静脉血管内皮细胞株(HUVEC)细胞凋亡的最适浓度和时间。方法:体外培养HUVEC,将Aβ1-42分为5×10-4、5×10-3、5×10-2、5×10-1、5、5×10、5×102μmol/L 7个浓度,分别刺激0.5、2、6、12、24、48 h后,采用CCK-8法观察细胞活力,荧光显微镜观察Hochest33342/PI双染后细胞形态。结果:同阴性组比较,Aβ1-42浓度≥5μmol/L刺激48h均可引起细胞活力明显下降(P<0.05)。5×10,5×102μmol/L刺激24 h可引起HUVEC细胞活力下降,但48hHUVEC细胞活力下降更明显(P<0.05)。Hochest33342/PI双染可见明显核浓缩和凋亡小体的产生,甚至出现死亡现象。结论:Aβ1-42刺激后引起HUVEC细胞活力下降呈时间与剂量依赖性,综合时间与剂量考虑,5×10μmol/L刺激24h可致理想的HUVEC细胞凋亡模型。Objective： To screen the optimal concentration and incubation time of β-amyloid 1-42 in inducing apoptosis of Human umbilical vein endothelial cells（HUVEC）. Methods： HUVEC were cultivated in vitro. β-amyloid 1-42 was diluted into 5 × 10-4,5 × 10-3,5 × 10-2,5 × 10-1,5,5 × 10,5 × 102μmol/L;the incubation time was set at 0.5,2,6,12,24 and 48 h. The cell viability was observed by CCK-8 and the cell morphology was observed by fluorescence microscopy after dyeing with Hochest33342/PI. Results： Compared with negative control group, cell viability of all the groups with Aβ1-42≥ 5μmol/L after stimulating 48 hours was decreased significantly（P〈 0.05）. The viability of HUVEC incubated with 5 × 10 and 5 × 102μmol/L β-amyloid1-42 for 24 hours or 48 hours was decreased significantly（P 〈 0.05）. The nuclear enrichment and apoptotic body could be o bserved by Hochest33342/PI. Conclusion： HUVEC cell viability decreased in a time and dose related way after incubated with β-amyloid1-42. Considering of the time and dose comprehensively,incubation with 5 × 10 μmol/L β-amyloid 1-42 for 24 hours can establish the ideal model of HUVEC apoptosis.

关 键 词：β样淀粉肽1-42 HUVEC细胞 细胞凋亡 

分 类 号：R967[医药卫生—药理学]