阻塞性睡眠呼吸暂停模式间歇低氧对人脐静脉内皮细胞中内皮素-1、一氧化氮及一氧化氮合酶的影响  

Effects of obstructive sleep apnea style intermittent hypoxia on endothelin-1, nitric oxide, and nitric oxide synthase in endothelium:experiment with human umbilical vein endothelial cells

作　　者：赵海燕[1] 陈宝元[1] 曹洁[1] 冯靖[1] 郭美南[1] 

机构地区：[1]天津医科大学总医院呼吸科,300052

出　　处：《中华医学杂志》2007年第31期2189-2192,共4页National Medical Journal of China

摘　　要：目的通过观察不同间歇低氧(IH)模式下人脐静脉内皮细胞中内皮素-1(ET-1)浓度、一氧化氮(NO)浓度、一氧化氮合酶(NOS)活性及内皮型一氧化氮合酶(eNOS)mRNA 表达,探讨内皮功能障碍在阻塞性睡眠呼吸暂停综合征合并高血压发病机制中的作用。方法采用人脐静脉内皮细胞可传代细胞株 ECV304细胞系,暴露于不同的低氧/再氧合条件。暴露完成后测定 ET-1浓度(酶免疫测定,EIA)、一氧化氮浓度(硝酸还原酶法)、NOS 活性(化学比色法)及 eNOS mRNA 表达(RT-PCR)。结果间歇低氧组与间歇正常氧组及空白对照组比较,ET-1浓度升高(F=28.453,P=0.000),NO 浓度降低(F=65.252,P=0.000),NOS 活性(F=5.969,P=0.008)及 eNOS mRNA 表达下降(F=16.630,P=0.000)。相同累加低氧时间及程度的间歇低氧组与持续低氧组比较 ET-1浓度升高(t=2.742,P=0.024),NO 浓度降低(t=3.347,P=0.004),NOS 活性(t=3.989,P=0.004)及eNOS mRNA 表达下降(t=5.045,P=0.000);且不同频率间歇低氧组随再氧合时间的延长,ET-1浓度呈增加趋势(F=213.254,P=0.000),而 NO 浓度(F=15.747,P=0.000)、NOS 活性(F=2.933,P=0.048)及 eNOS mRNA 表达下降(F=5.954,P=0.002),但当再氧合时间过长时 ET-1浓度下降,NO 系统各指标上调。结论间歇低氧的低氧及再氧合过程均可导致内皮功能障碍、NO 和 ET-1释放失衡,从而促进了阻塞性睡眠呼吸暂停相关心血管疾病的发生和发展。Objective To investigate the effects of intermittent hypoxia （IH） on the expression of endothelin-1 （ ET-1 ） , nitric oxide（NO） ,and nitric oxide synthase（NOS） in endothelium ,and to evaluate the role of functional disorder of endothelium in the mechanism of obstructive sleep apnea syndrome （OSAS） induced hypertension. Methods Human umbilical vein endothelial cells （HUVECs） of the line ECV304 were cultured and divided into 4 groups： IH group （exposed to 1.5% 02 for 15 s and 21% 02 for 1 min 15 s, 3 rain 45 s, 5 min 15 s, or 8 min 15 s respectively alternatively with 60 episodes）, intermittent normal oxygen group （exposed to 21% 02 for 15 s and 225 s alternatively with 60 episodes）, continuous hypoxia group （exposed to 1.5% for 15 min）, and blank control group. Then the culture fluid was collected. The levels of activity of ET-1 （EIA） NO, and NOS were detected by enzyme immune assay, nitric acid recluctase method, and chemical colorimetric analysis respectively. RT-PCR was used to detect the mRNA expression of endothelial NOS （ eNOS）. Results Compared with those of the intermittent normal oxygen group and blank control groups the ET-1 levels of the 4 IH sub groups were significantly higher （ F = 28. 453, P = 0. 000） , the NO levels ere significantly lower F = 65. 252, P = 0. 000）, the NOS activity levels were significantly lower （ F = 5. 969, P = 0. 008 ）, and eNOS mRNA was significantly down-regulated （ F = 16.630,P = 0. 000）. Compared with continuous hypoxia group with the exposure time equivalent to the accumulated hypoxia time （ 15 s with 60 episodes） of the IH group, the ET-1 level was significantly higher （t =2. 742 ,P =0. 024）, the NO level was significantly lower（ t = 3. 347 ,P =0. 004）, the NOS activity level was signitlcantly lower （ t = 3.989, P = 0. 004 ）, and the eNOS mRNA expression was down-regnlated （ t = 5. 045,P = 0. 000）. In the different IH group along with the prolongation of the re-oxygenation time from 105 s to 4

关 键 词：睡眠呼吸暂停 阻塞性 内皮缩血管肽类 一氧化氮 间歇低氧 

分 类 号：R766[医药卫生—耳鼻咽喉科]