LPS诱导HUVEC细胞凋亡最适浓度与时间选择

目的：探讨脂多糖诱导HUVEC细胞凋亡的最适浓度和时间。方法：体外培养人脐静脉血管内皮细胞株（HU—VEC），将脂多糖（LPS）分为100、10、1、10^-1、10^-2、10^-3、10^-4、10^-5μg·ml^-18个浓度，分别刺激0．5、1、2、6、12、24h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：同阴性对照组比较，药物浓度≥10^-2μg·ml^-1刺激24h均可引起细胞活力明显下降（P〈0．05）。100μg·ml^-1刺激1h，10μg·ml^-1与1μg·ml^-1刺激6h可引起HUVEC细胞活力下降，但12h与24hHUVEC细胞活力下降更明显（P〈O．05），10^-1μg·ml^-1刺激12h可引起HUVEC细胞活力下降（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：LPS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，10^-1μg·ml^-1刺激12h或24h与10^-2μg·ml^-1刺激24h可致理想的HUVEC细胞凋亡模型。

Study on the optimal concentration and time of LPS inducing the apoptosis of HUVEC cells

Objective.. To explore the optimal concentration and time of LPS inducing the apoptosis of HUVEC cells. Methods： Human umbilical vein endothelial cells （HUVEC） were cultivated in vitro, Lipopolysaccharides were divided intol00.10.1.10^-1, 10^-2 .10-3 .10-4 .10-5μg . ml^-1 respectively, time of stimulate were 0.5, 1, 2, 6, 12, 24 h, cell viability were observed by CCK-8~ cell morphology were measured by fluorescence microscopy after dyeing with Hochest33342/PI. Results： Compared with control group, cell viability was sigrdficantly decreased in over 10-2 g. ml^-1 LPS and 24 h（P〈0.05）. 100 /lg . ml^-1 LPS for 1 h, 10 μg . ml^-1 and 1 g . ml^-16 h can induce HLrVEC cell viability decreased, but 12 h and 24 h cell viability decreased more significantly （P〈0.05）, 10-1 g . ml-1LPS 12 h can induce HUVEC cell viability decreased （P〈0. 05）; nuclear enrichment and apoptotic body, and even death can be obviously by Hochest33342/PI. Conclusion： HUVEC celt viability decrease after administrate with LPS in a time and dose way, comprehensive consider time and dose, 1 g . ml^-1 LPS stimulate 12 h or 24 h and 10-1 g . ml^-1 LPS stimulate 24 h can cause the ideal model of HUVEC apoptosis.