核因子-κB在肺气肿合并间歇低氧大鼠内皮细胞炎症损伤的机制  

The role of nuclear factor-κB in endothelial cell inflammatory injury by intermittent hypoxia in rat with emphysema

作　　者：梁磊[1] 陈怀永[2] 吴琦[3] 孙昕[3] 冯靖[4] 郑兴杰[3] 李雪[2] 徐龙[2] 

机构地区：[1]天津医科大学,300070 [2]天津市海河医院基础实验室 [3]天津市海河医院呼吸科 [4]天津医科大学总医院呼吸与危重症医学科

出　　处：《中华结核和呼吸杂志》2015年第3期196-201,共6页Chinese Journal of Tuberculosis and Respiratory Diseases

基　　金：国家自然科学基金（81270144）;天津市应用基础与前沿技术（13JCYBJC22400）

摘　　要：目的 探讨肺气肿合并间歇低氧对大鼠核因子-κB及其下游基因产物表达影响及核因子-κB在主动脉内皮炎症损伤及功能损害的作用机制.方法 60只雄性Wistar大鼠按随机数字表法分为4组,每组15只：对照组常规饲养;肺气肿组每天常氧下熏烟2次,每次30 min;间歇低氧组每天间歇低氧暴露8h;肺气肿合并间歇低氧组每天常氧熏烟2次,每次30 min,并间歇低氧暴露8h.各组均暴露8周,第9周初每组随机取5只大鼠测量血气;剩余大鼠处死,取肺组织,HE染色光镜下观察肺组织病理形态.随后取胸主动脉内皮组织,采用ELISA测定内皮素-1、肿瘤坏死因子（TNF）-α和IL-8水平,硝酸还原酶法测一氧化氮水平,RT-PCR测定核因子-κB mRNA,细胞间黏附分子（ICAM）-1mRNA,基质金属蛋白酶（MMP）-9 mRNA,内皮型一氧化氮合酶（eNOS） mRNA表达水平.结果 肺组织病理形态和血气分析结果证明,肺气肿合并间歇低氧大鼠模型建立成功.肺气肿合并间歇低氧组主动脉内皮组织内皮素-1、TNF-α和IL-8含量分别为（172.4＆#177;1.6）、（104.1＆#177;1.4）和（272.1＆#177;3.6） ng/L,与对照组、肺气肿组和间歇低氧组相比均明显增高（均P＜0.05）.一氧化氮水平为（27.07＆#177;0.57） μmol/L,与对照组、肺气肿组和间歇低氧组相比较低（均P＜0.05）.肺气肿合并间歇低氧组主动脉内皮组织核因子-κB mRNA、ICAM-1 mRNA、MMP-9 mRNA表达升高水平与对照组、肺气肿组和间歇低氧组相比较高（均P ＜0.05）.eNOS mRNA表达水平较正常对照组、肺气肿组、间歇低氧组均明显降低（P＜0.05）,核因子-κB mRNA表达水平与MMP-9 mRNA水平呈正相关（r=0.572,P＜0.01）,核因子-κB mRNA与eNOS mRNA水平呈负相关（r=-0.534,P＜0.01）;肺气肿组核因子-κB mRNA,eNOS mRNA表达水平与间歇低氧组比较差异无统计学意义（P＞0.05）.结论 肺气肿合并间歇低氧暴露相对于单纯肺气肿或间歇低氧暴Objective To investigate mechanism underlying the role of nuclear factor Kappa B （NF-κB） which induced inflammatory injury and functional lesions of aortic endothelial cells in rat with emphysema and intermittent hypoxia.Methods Sixty male Wistar rats were divided randomly into 4 experimental groups （n =15 each group）：control group,emphysema group,intermittent hypoxia （IH） group,emphysema with intermittent hypoxia group.The rats in control group had ad libitum access to food and water under normal circumstance.The rats in the emphysema group were exposed to cigarette smoke twice daily （30 min each time）.As for IH group,the rats were exposed to intermittent hypoxia circumstance （8 h/day）.Both cigarette smoke twice a day （30 min each time） and intermittent hypoxia circumstance （8 h/day） were imposed on the rats in emphysema with intermittent hypoxia group.All the rats were exposed for 8 weeks.Five rats were randomly selected from each group to measure the blood gas on the ninth week.We collected lung and endothelial tissues of thoracic aorta from the rest sacrificed rats,and observed the pathological changes of lung tissue through HE staining.The levels of ET-1,TNF-α and IL-8 in rat endothelial tissues of thoracic aorta were measured by ELISA testing.Nitrate reductase was used to measure the levels of NO,and RT-PCR to detect the levels of NF-κB mRNA,ICAM-1 mRNA,MMP-9 mRNA and eNOS mRNA.Results Lung pathology and blood gas results showed that the rat model of emphysema with intermittent hypoxia was established successfully.The levels of ET-1,TNF-α,IL-8 in emphysema with intermittent hypoxia group were （172.4 ＆#177; 1.6） ng/L,（104.1 ＆#177; 1.4） ng/L,（272.1 ＆#177; 3.6） ng/L respectively,significantly higher than the control group,emphysema group and intermittent hypoxia group （all P〈0.05）.The level of NO was （27.07 ＆#177;0.57） μmol/L,which was significant reduced; the expression of NF-κB mRNA,ICAM-1 mRNA,MMP-9 mRNA in emphysema with intermittent hypoxia

关 键 词：肺气肿 间歇低氧 重叠综合征 

分 类 号：R563.3[医药卫生—呼吸系统]