Preconditioning of carbon monoxide releasing molecule-derived CO attenuates LPS-induced activation of HUVEC.
Preconditioning of carbon monoxide releasing molecule-derived CO attenuates LPS-induced activation of HUVEC.
复制标题
一氧化碳释放分子衍生的 CO 的预处理可减弱 LPS 诱导的 HUVEC 激活。
DOI：
10.7150/ijbs.4.270
复制
发表时间：
2008-08-22
影响因子：
9.2
通讯作者：
Shi G
中科院分区：
文献类型：
--
作者：
Sun B;Zou X;Chen Y;Zhang P;Shi G
目的：探讨CORM-2释放的CO对脂多糖(LPS)诱导的内皮细胞(HUVEC)活化的影响及其机制。方法：人脐静脉内皮细胞用50或100μM的CORM-2处理2小时，再用10μg/ml的内毒素清洗和刺激4小时。通过检测细胞内DHR 123的氧化或DAF-FM、特定的H_2O_2和NO荧光素的硝化来评价HUVEC的激活(氧化应激)。检测HO-1、诱导型一氧化氮合酶(Western印迹)和细胞间黏附分子-1(Cell ELISA)蛋白的表达及炎症相关转录因子核转录因子B(NF-κB，EMSA)的激活情况。此外，还检测了PMN与HUVEC的粘附性。结果：CORM-2可抑制内皮细胞产生ROS和NO，在蛋白水平上上调HO-1的表达，减少诱导型一氧化氮合酶的表达，抑制内毒素诱导的NF-κB的活化，下调ICAM-1的表达，减少中性粒细胞与内毒素刺激的内皮细胞的黏附。结论：CORM-2释放的CO的预适应通过干扰细胞内氧化应激的诱导而产生抗炎作用。此外，它还支持CO是诱导型一氧化氮合酶和核因子-κB的有效抑制剂的观点。
Objective: To investigate the effects and potential mechanisms of preconditioning of tricarbonyldichlororuthenium (III) dimer (CORM-2)-liberated CO on LPS-induced activation of endothelial cells (HUVEC). Methods: HUVEC were pretreated with CORM-2 at the concentration of 50 or 100μM for 2 hrs, washed and stimulated with LPS (10μg/ml) for additional 4 hrs. Activation (oxidative stress) of HUVEC was assessed by measuring intracellular oxidation of DHR 123 or nitration of DAF-FM, specific H2O2 and NO fluorochromes, respectively. The expression of HO-1, iNOS (Western blot) and ICAM-1 (cell ELISA) proteins and activation of inflammation-relevant transcription factor, NF-κB (EMSA) were assessed. In addition, PMN adhesion to HUVEC was also assessed. Results: The obtained data indicate that pretreatment of HUVEC with CORM-2 results in: 1) decrease of LPS-induced production of ROS and NO; 2) up-regulation of HO-1 but decrease in iNOS at the protein levels; 3) inhibition of LPS-induced activation of NF-κB; and 4) downregulation of expression of ICAM-1, and this was accompanied by a decrease of PMN adhesion to LPS-stimulated HUVEC. Conclusions: Preconditioning of CO liberated by CORM-2 elicited its anti-inflammatory effects by interfering with the induction of intracellular oxidative stress. In addition, it also supports the notion that CO is a potent inhibitor of iNOS and NF-κB.