摘要
目的:探讨过氧化氢(H2O2)对肺动脉内皮细胞环氧合酶-2(COX-2)表达的影响及钙-钙调蛋白激酶Ⅱ(CaMKⅡ)在其中的作用。方法:采用活细胞计数法(CCK-8法)检测H2O2处理肺动脉内皮细胞后的细胞活性,采用RT-PCR检测COX-2 mRNA的表达水平,采用Western blotting检测COX-2蛋白质的表达水平。结果:H2O2增强COX-2表达,呈浓度和时间依赖性。100μmol/L H2O2处理肺动脉内皮细胞4 h,COX-2 mRNA和蛋白质表达水平明显高于正常对照组,COX-2 mRNA水平为正常对照组的256.01%±22.36%(P<0.05),蛋白质水平为正常对照组的216.65%±21.52%(P<0.05)。CaMKⅡ特异性抑制剂KN-93能抑制H2O2的这一效应。结论:H2O2可增强肺动脉内皮细胞COX-2基因的表达,CaMKⅡ是H2O2增强肺动脉内皮细胞COX-2基因表达的途径之一。
AIM： To investigate if hydrogen peroxide may alter COX -2 gene expression in pulmonary artery endothelial cells （PAECs） and how CaMK 11 functions in this process. METHODS： Cultured pulmonary arterial endothelial cells were treated with different concentrations of hydrogen peroxide for different durations. The cells survival rates were measured by CCK - 8 after the cells were treated by hydrogen peroxide. The level of COX - 2 mRNA and protein were measured by RT - PCR and Western blotting, respectively. RESULTS ： The results showed that hydrogen peroxide up - regulated COX- 2 mRNA and protein levels in a concentration - and time -dependent manners. Incubation with 100 μmol/L H2O2 for 4 h increased COX - 2 mRNA and protein level to 256. 01% ±22. 36% （P 〈 0. 05） and 216.65%±21.52% （P 〈0. 05）, respectively. The up- regulatory effects were prevented by KN -93, a selective CaMK Ⅱ inhibitor, at concentration of 10 μmol/L. CONCLUSION： These results suggest that hydrogen peroxide up -regulates COX - 2 gene transcription in PAECs by a signaling pathway involving CaMK Ⅱ.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第10期1968-1972,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30200099No.30570793)
