摘要
目的研究慢性间歇低氧(CIH)大鼠中性多核白细胞(PMNs)与内皮细胞间接共培养C反应蛋白(CRP)、肿瘤坏死因子-α(TNF-α)和细胞间黏附分子-1(ICAM-1)水平的变化,探讨炎症因子水平的改变对内皮细胞的影响及抗氧化剂4-羟基-2,2,6,6-四甲基哌啶(Tempol)的干预作用。方法 36只雄性Wistar大鼠采用随机数字表法分为常氧对照组(NC组)、间歇低氧组(IH组)、间歇低氧Tempol干预组(IHT组)和间歇低氧盐水干预组(IHN组),每组9只。除NC组外均给予间歇低氧环境,IHT组低氧暴露前给予100 mg/(kg·d)的Tempol腹腔注射,生理盐水稀释,IHN组同时给予等体积生理盐水腹腔注射。暴露6周后处死大鼠,分离纯化PMNs。PMNs与正常大鼠主动脉内皮细胞Transwell间接共培养4 h,酶联免疫吸附法(ELISA)测定间接共培养上、下室上清中CRP、TNF-α和ICAM-1的水平。结果 NC组、IH组、IHT组和IHN组上、下室CRP、TNF-α和ICAM-1水平比较,差异均无统计学意义(均P>0.05);与NC组下室相比,IH组和IHN组下室CRP、TNF-α和ICAM-1水平均明显升高,差异有统计学意义(P<0.05);与IH组和IHN组下室比较,IHT组下室CRP、TNF-α和ICAM-1水平均明显降低,差异有统计学意义(均P<0.05);IHT组下室与NC组下室相比,CRP、TNF-α和ICAM-1水平差异均无统计学意义(均P>0.05);IH组下室与IHN组下室比较,以上指标差异均无统计学意义(均P>0.05)。结论 CIH大鼠体内的PMNs与内皮细胞间接共培养,可上调CRP、TNF-α和ICAM-1等炎症因子水平,可能在CIH引起内皮损伤中发挥重要作用;抗氧化剂Tempol可能阻断炎症反应,干预CIH引起的内皮功能障碍的发生。
[Objectives] To study the changes of CRP, TNF-α and ICAM-1 induced by indirectly cocul- turing rat aortic endothelial cells and PMNs from rats exposed to chronic intermittent hypoxia （CIH） and to investigate the role of inflammatory cytokines on rat aortic endothelial cells and the effect of 4-hydroxy-2, 2, 6, 6-tetramethylpiperidine-N-oxyl or 4-hydroxy-TEMPO. [Methods] A CIH animal model of rats was estab-lished to mimic the intermittent hypoxia/re-oxygenation （IH/ROX） of obstructive sleep apnea syndrome in hu- mans. Thirty-six healthy male Wistar rats were randomly assigned to 4 groups： normoxic control group （NC group）, intermittent hypoxia group （IH group）, intermittent hypoxia tempol treatment group （IHT group）, inter- mittent hypoxia normal saline matched group （IHN group）, with 9 rats in each group. IHT group was treated with 10% Tempol 100 mg/kg^-1·d^-1 by intraperitoneal injection before exposed to IH, while IHN group was treated with NS as controls. After the experiment, PMNs was isolated, normoxic aortic endothelial cells and PMNs coincubated indirectly using Transwell for 4 hours. The levels of CRP, TNF-α and ICAM-1 in the up- per and lower chamber of Transwell were measured. [Results] The levels of CRP, TNF-α and ICAM-1 in the upper chamber of Transwell between the NC group, IH group, IHT group and IHN group were not signif- icantly different as compared to those in the lower chamber of Transwell （all P 〉0.05）. Compared with the lower chamber of NC group, in the lower chamber of IH group and the IHN group, the levels of CRP, TNF- α and ICAM-1 were increased （all P〈0.05）. Compared with the lower chamber of the IHN group and the CIH group, in the lower chamber of IHT group, the levels of CRP, TNF-α and ICAM-1 were decreased （all P〈0.05）. The levels of CRP, TNF-α and ICAM-1 in the lower chamber of IHT group were not significantly different compared to the NC group （all P〉0.05）. The levels of CRP, TNF-α and ICAM-1 in the lower chamber of IHN group were not significantly different as compared to those in the IH group （all P 〉0.05）. [Conclusion] Indirectly eoculturing PMNs and endothelial cells could up-regulate the level of TNF-α, CRP and ICAM-1 and the high levels of those inflammation cytokines could cause rat endothelial cells injury. Tempol could prevent CIH-indueed endothelial dysfunction by its anti-inflammation effect.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2014年第2期5-9,共5页
China Journal of Modern Medicine
基金
国家自然科学基金(No:81170071)
