摘要
目的:通过检测间歇低氧刺激下人脐静脉内皮细胞(HUVECs)的凋亡率、炎性因子及氧化应激因子的水平,探讨不同间歇低氧暴露时间对HUVECs损伤的影响。方法:分别给予HUVECs间歇低氧刺激(1%O25 min;21%O210 min)及常氧(对照组)处理。在不同的间歇低氧暴露时间点应用膜联蛋白(annexin)Ⅴ-异硫氰酸荧光素(FITC)/碘化丙啶(PI)检测细胞凋亡率,酶联免疫吸附试验(ELISA)法测定细胞上清液中白介素(IL)-6和IL-8水平,分光光度法检测上清液中丙二醛(MDA)水平。结果:间歇低氧处理12、16、20和24 h的细胞凋亡率逐渐增高,且均高于对照组(P<0.01),间歇低氧处理的细胞上清液中IL-6水平均显著高于对照组(P<0.01),在4、8、12、16 h各时间点逐渐升高,并于16 h达到高峰[(251.40±49.45)ng/L]。间歇低氧处理8、12、16、20和24 h的细胞上清液内的IL-8和MDA水平均明显高于对照组(P<0.01),且分别于20 h和16 h达到峰值[IL-8:(737.70±31.08)ng/L;MDA:(2.55±0.04)μmol/L]。结论:间歇低氧刺激可导致HUVECs凋亡增加,炎症反应和氧化应激水平升高,在间歇低氧刺激一定时间后达到高峰。
Objective To study the effect of different duration of exposure to intermittent hypoxia (IH) on apoptosis,inflammatory eytokines and oxidative stress of human umbilical vein endothelial cells (HUVECs). Methods HUVECswere treated with intermittent hypoxia (exposed to 1% O2 for 5 min and 21% O2 for 10 min) for 12, 16, 20, 24 h or undernormoxia (control group). At different time points, apoptosis rates of HUVECs were determined by annexin V-fluoresceinisothiocyanate (FITC)/propidium iodide (P I); levels of interleukin (IL)-6 and IL-8 in culture supernatant were detected byenzyme linked immunosorbent assay (ELISA), and malondialdehyde (MDA) were measured by speetrophotometry. ResultsThe apoptosis rates of HUVECs under IH for 12, 16, 20, 24 h were significantly higher than that of control group (P〈0.01)and were increased gradually. The IL-6 levels in culture supernatant from IH groups were higher than the control group(P〈0.01); IL-6 levels were increased gradually from 4, 8, 12, 16 h and reached the peak at 16 h [(251.40±49.45) ng/L].Higher levels of IL-8 and MDA were found in 8, 12, 16, 20, 24 h IH groups than that in control group (P〈0.01), andreached the peak at 20 h and 16 h, respectively [IL-8 (737.70±31.08) ng/L; MDA: (2.55±0.04) μmol/L]. ConclusionsIntermittent hypoxia led to increased apoptosis of HUVECs. Inflammation and oxidative stress increased under IH, andreached the peak after a certain period of exposure.
出处
《内科理论与实践》
2014年第2期121-125,共5页
Journal of Internal Medicine Concepts & Practice
基金
国家自然科学基金项目(项目编号:81070068)
上海市科委基础研究重点项目(项目编号:11JC1411302)
关键词
阻塞性睡眠呼吸暂停低通气综合征
间歇低氧
暴露时间
炎症
氧化应激
Obstructive sleep apnea-hypopnea syndrome
Intermittent hypoxia
Exposure duration
Inflammation
Oxidative stress